This study aims to investigate the effects of Japanese encephalitis virus(JEV)on TLRs signaling pathway and its regulation of the secretion of inflammatory factors during the infection of testicular interstitial cells,In this study,the mRNA levels of TLR3,TLR7,TLR8,TRIF and MyD88 genes were detected by qPCR after 1 MOI dose of JEV was inoculated into testicular stro-mal cells at different time periods.Western blot assay was used to detect the expression levels of TLR3,TLR7,TRIF and MyD88 protein at 6 h after JEV infection,and ELISA was used to detect the expression levels of IL-1β,IL-6 and TNF-α at different time periods(6,12 and 24 h).The re-sults showed as follows:After 6 h of JEV infection,the mRNA levels of TLR3,TLR7,TRIF and MyD88 genes were significantly up-regulated(P＜0.05),and the mRNA levels of TLR8 genes were down-regulated(P＜0.05).Western blot results showed that the protein expressions of TLR3,TLR7,TRIF and MyD88 were significantly up-regulated when JEV infected testicular stromal cells for 6 h(P＜0.05),which was consistent with the corresponding mRNA transcription levels.There was no significant change in TLR8 protein expression.ELISA results showed that 6 h after JEV infection of testicular stromal cells,IL-6 was significantly increased(P＜0.01),and the expressions of IL-1β and TNF-α were not changed.TLR3,TLR7,TLR8,TRIF and MyD88 were si-lenced by siRNA,and the silenced cells were inoculated with JEV for 6 h,and IL-6 expression lev-els were detected by ELISA.The results showed that silenced TLR3,TLR7,TLR8,TRIF and MyD88 could significantly reduce the increase of IL-6 secretion induced by JEV infection(P＜0.05).These results indicated that JEV could induce the expression of inflammatory factor IL-6 by activating TLR3,TLR7 and TLR8 signaling pathway after infection of testicular stromal cells.This study provides a reference for further elucidating the mechanism of reproductive disorders caused by JEV infection.

Porcine circovirus type 2(PCV2)is the main pathogen causing porcine circovirus related diseases.PCV2 infection in pigs may lead to porcine dermatitis and nephrotic syndrome(PDNS)and weaned piglets multiple system failure syndrome(PMWS),etc.At present,the pathogenic mechanism is not fully understood.PCV2 is a single strand of negative link DNA,which can cause immune suppression in the body and lead to increased secondary susceptibility,which has a syner-gistic effect with various pig diseases and brings major economic losses to the pig industry.Al-though there are commercial vaccines,the prevention of vaccines has certain limitations and there is no effective drug treatment so far,an outbreak will threaten people's life and health and public safety,resulting in significant economic losses.In order to understand the latest progress of PCV2 escape mechanism and prevention and control,this paper summarizes the inhibition of interferon production,regulation of apoptosis,regulation of autophagy,regulation of pyroptosis and inflam-matory response,evasion of adaptive immune response,and prevention and control of PCV2,in or-der to provide new theoretical ideas for the research and prevention and control of PCV2.

Breast cancer is the most common cancer among women worldwide, and it seriously threatens women′s life and health. circular RNA (circRNA) play a key role in the development and drug resistance of various cancers, including breast cancer. circRNA is an endogenous RNA molecule with a single-stranded closed structure, which has unique biological characteristics and function, and is a current research hotspot. This article will review circRNA as potential biomarkers to predict breast cancer drug resistance and therapeutic targets.

【Objective】 To study the effect of air inlet modes on the temperature variation, fluctuation, uniformity and coefficient of variation(CV), so as to evaluate the stability and uniformity of the temperature field in refrigerated warehouse for blood. 【Methods】 The temperature changes of blood refrigerated warehouse under different modes of air inlet during compressor operation were analyzed. The stability of the temperature field in the storehouse was evaluated by the changes, fluctuation, uniformity, CV and deviation of temperature at each testing point. 【Results】 The average value of temperature in the storehouse, adopting air inlet via straight blow, was (4.98±0.92)℃, while that of air inlet via micro-hole mode was(4.15±0.25)℃, with significant differences between each other(P<0.05). As to the CV of temperature, air inlet via straight blow was significantly different from that via micro hole(P<0.01). The fluctuation, uniformity and deviation of temperature created by straight blow and micro hole were 1.85±1.11 vs 0.49±0.38, 1.00±0.68 vs 0.47±0.37, and 0.61±0.45 vs 0.27±0.21, respectively, with significant differences between each other(P<0.01). 【Conclusion】 Compared with straight blow, the mean temperature created by micro hole was closer to the median value (4℃) of the temperature range, i. e.(4±2)℃, during blood storage. Otherwise, micro hole demonstrated a smaller CV of temperature, and superior performance in fluctuation, uniformity and deviation of the temperature at the testing points, which was conducive to ensure the stability of storehouse temperature field.

Objective:To investigate the association between serum thyroid level and prognosis of critically ill children with euthyroid sick syndrome(ESS).Methods:The clinical data and serum thyroid hormone levels of 176 children with ESS who were admitted to the Department of Pediatric Intensive Care Medicine at West China Second Hospital of Sichuan University from January 2015 to April 2021 were retrospectively collected.According to the prognosis, the children were divided into improved group and invalid group, as well as basic disease group and non basic disease group, and the differences of thyroid hormone between two groups were compared.The pediatric risk of mortality Ⅲ(PRISMⅢ) scores within 24 hours of admission were assessed, and the correlation between thyroid hormone level and PRISMⅢ score was analyzed.Results:Among 176 critically ill children with ESS, the most common diseases were sepsis(31.8%), severe pneumonia (23.8%) and heart failure(10.7%), respectively.The levels of free T3(FT3), T3, free T4(FT4) and T4 in invalid group were significantly lower than those in improved group ( P<0.05), but there was no statistical difference in thyroid-stimulating hormone(TSH) level between two groups( P>0.05). The levels of FT3, T3, FT4 and T4 were negatively correlated with PRISMⅢ score( r=-0.419, -0.459, -0.341, -0.383, respectively, P<0.05), and there was no correlation between TSH level and PRISMⅢ score ( P>0.05). The common underlying diseases of severe children with ESS were malnutrition(31/98), heart disease(30/98), hematologic neoplasms(15/98), and bronchopulmonary dysplasia(10/98). The median age of children in basic disease group was younger than that in non-basic disease group(0.7 years old vs. 2.0 years old, P<0.05); The proportion of children with underlying diseases in invalid group was 24.5%, which was significantly higher than that of children without underlying diseases (6.4%), and the difference was statistically significant ( P<0.05); There were no significant differences in the levels of FT3, T3, FT4, T4 and TSH between two groups ( P>0.05). Conclusion:In critically ill children, a variety of diseases can lead to ESS, and sepsis is the most common disease.Young children with underlying diseases should be more alert to ESS.The more severe the disease, the greater the decline of FT3, T3, FT4 and T4 levels.When low T3, T4 and TSH occur simultaneously, the prognosis of the children may be worse.Thyroid hormone level could be used as an indicator to evaluate the prognosis of critically ill children, which is needed further studies to explore.

AIM: To establish an LC-MS/MS method to determine the concentration of ambrisentan in human plasma and apply it to the study of human pharmacokinetics. METHODS: After extracting ambrisentan and internal standard from human plasma by liquid-liquid extraction, chromatographic separation was performed on a Waters Symmetry C

Objective To summarize the clinical characteristics of children with Kawasaki disease (KD) in pediatric intensive care unit(PICU). Methods Medical record of children with KD at PICU were collected. At the same time,29 cases of KD in PICU were 1∶3 matched by age,gender and the time admitted in hospital with those admitted in general pediatric department(control group). Results PICU patients had longer length of hospital stay,longer fever duration compared with control group. In addition,patients in PICU had higher neutrophil percentage,C reaction protein,creatinine,urea nitrogen,N￣terminal natriuretic peptide and procalcitonin,but lower hemoglobin,blood platelet and albumin compared with the control group. What′s more,patients in PICU tended to find hemoglobin<100 g/L,platelet<150×109/L,albumin<30 g/L,abnor￣mal in urine routine and echocardiographic and more likely to have fever longer than 10 days when used intra￣venous immunoglobulin(IVIG) compared with control group. And PICU patients were more likely to require therapy with antibiotics,albumin,glucocorticoid and the second dose of IVIG. Some part of children in PICU group were treated with IVIG and glucocorticoid because of doubted severe infection before KD diagnosed,all patients in the control group used IVIG after the diagnosis. Conclusion Patients who admitted in PICU are severe and not typical in clinical manifestation. These patients are easily misdiagnosed as sepsis and more likely to be IVIG￣refractory and have coronary artery damage. We still worry that somebody might be misdi￣agnosed as sepsis,who are treated with IVIG and get better. Because they are not diagnosed as KD,these pa￣tient would not followe up like KD,but have potential risk of cardiovascular disease and need more alarming.

Objective To study the survival status of very elderly patients with intertrochanteric fracture admitted in hospital and its risk factors.Methods Clinical data of 54 patients with intertrochanteric fracture aged 90 years and over admitted in hospital from Jan.2006 to Dec.2013 were retrospectively analyzed.5 patients were lost to follow-up.The survival status of patients was analyzed.Results Patients were followed up until January 2015.The period of follow-up ranged from 85 months to 0.2 months,with an average follow-up of 33.3 months.17 cases were survived.Harris scores were(61.2 ± 10.6).The overall 1-,3-,5-year survival rates were(0.88 ± 0.05),(0.35 ±0.08),(0.18 ± 0.08),respectively.The mean and median survival times were (33.3 ± 4.6) months and(25.0±5.4) months.Cox proportional hazard regression model showed that American Society of Anaesthesiologists(ASA) score,operation time and the number of comorbidities were the risk factors for the survival time of very elderly patients with intertrochanteric fracture (all P ＜0.05).Conclusions Compared with conservative treatment,surgical treatment can increase the survival time of the elderly patients with intertrochanteric fracture.ASA score,operation time and the number of comorbidities may be the risk factors for the survival time of very elderly patients with intertrochanteric fracture.

Thymopentin (TP5) and bursopentin (BP5) are both immunopotentiators. To explore whether the TP5-BP5 fusion peptide (TBP5) has adjuvant activity or not, we cloned the TBP5 gene and confirmed that the TBP5 gene in a recombinant prokaryotic expression plasmid was successfully expressed in Escherichia coli BL21. TBP5 significantly promoted the proliferation of thymic and splenic lymphocytes of mice. The potential adjuvant activity of the TBP5 was examined in mice by coinjecting TBP5 and H9N2 avian influenza virus (AIV) inactivated vaccine. HI antibody titers, HA antibodies and cytokines levels (IL-4 and IFN-γ) were determined. We found that TBP5 markedly elevated serum HI titers and HA antibody levels, induced the secretion of both IL-4 and IFN-γ cytokines. Furthermore, virus challenge experiments confirmed that TBP5 contributed to inhibition replication of the virus [H9N2 AIV (A/chicken/Jiangsu/NJ07/05)] from mouse lungs. Altogether, these findings suggest that TBP5 may be an effective adjuvant for avian vaccine and that this study provides a reference for further research on new vaccine adjuvants.
Adjuvants, Immunologic ; pharmacology ; Animals ; Antibodies, Viral ; blood ; Cell Proliferation ; drug effects ; Influenza A Virus, H9N2 Subtype ; drug effects ; physiology ; Influenza Vaccines ; immunology ; Interferon-gamma ; immunology ; Interleukin-4 ; immunology ; Lymphocytes ; drug effects ; Mice ; Oligopeptides ; immunology ; Orthomyxoviridae Infections ; drug therapy ; Recombinant Fusion Proteins ; immunology ; Spleen ; cytology ; Thymopentin ; immunology ; Thymus Gland ; cytology ; Vaccines, Inactivated ; immunology ; Virus Replication

Objective:To explore the accuracy of CT three dimensional reconstruction technique for the diagnosis of tibia shaft fracture accompanied by ankle injury .Methods:Both X‐ray examination and three dimensional CT scanning were conducted for the 117 patients with closed tibia shaft fracture ,who went to Zhongshan Hospital of Fudan University ,before surgery .The numbers of patients ,who were diagnosed with tibia shaft fracture accompanied by ankle injury ,were compared between the two examination methods .Results:Nineteen patients with tibia shaft fracture were diagnosed with accompanied posterior malleolar fracture by X‐ray examination before surgery . However ,34 patients were diagnosed with accompanied posterior malleolar fracture by CT three dimensional reconstruction images . Thus , 15 more patients were diagnosed by CT three dimensional reconstruction technique than that by X‐ray imaging .Conclusions:Comparing with X‐ray examination , three dimensional CT reconstruction technique could increase the diagnosis accuracy for posterior malleolar fracture associated with tibia shaft fracture and reduce the rate of missed diagnosis .