Diphtheria Toxin/Human B-Cell Activating Factor Fusion Protein Kills Human Acute Lymphoblastic Leukemia BALL-1 Cells: An Experimental Study

Objective： This study aimed to express a fusion protein of diphtheria toxin and human B cell-activating factor （DT388sBAFF） in Escherichia coli （E. coli） and investigate its activity in human B-lineage acute lymphoblastic leukemia 1 cells （BALL-1）. Methods： A fragment of DT388sBAFF fusion gene was separated from plasmid pUC57-DT388sBAFF digested with Nde I and Xho I, and inserted into the expression vector pcold II digested with the same enzymes. Recombinants were screened by the colony polymerase chain reaction （PCR） and restriction map. The recombinant expression vector was transformed into BL21 and its expression was induced by isopropyl β-D-1-thiogalactopyranoside （IPTG）. The recombinant protein was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS-PAGE） and Western blot, and then purified by Ni2＋-NTA affinity chromatography. The expression level of B cell-activating factor receptor （BAFF-R） on BALL-1 cells was assessed by real-time PCR. The receptor binding capacity of recombinant protein was determined by cell fluorescent assay. The specific cytotoxicity of recombinant protein on BALL-1 cells was detected by 3-（4,5-dimethylthiazolyl-2）-2,5-diphenyltetrazolium bromide （MTT） assay. Results： The expression level of recombinant protein was 50% of total bacterial proteins in E. coli, and the recombinant protein could bind to BAFF-R-positive BALL-1 cells and thereby produce a cytotoxic effect on the cells. Conclusion： The fusion protein expression vector DT388sBAFF was successfully constructed and the recombinant protein with selective cytotoxicity against BALL-1 cells was obtained, providing foundation for further study of the therapy of human B-lineage acute lymphoblastic leukemia.

Change of serum B-cell activating factor level in patients with positive antiphospholipid antibodies and previous adverse pregnancy outcomes and its significance

Background:B-cell activating factor(BAFF)is vital for B cell survival.Serum BAFF levels are elevated in thrombotic antiphospholipid syndrome,but little is known about levels in patients with positive antiphospholipid antibodies(aPLs)and previous adverse pregnancy outcomes(APOs).We aimed to analyze serum BAFF concentrations of these patients in early pregnancy along with different pregnancy outcomes.Methods:Thirty-six pregnant patients positive for aPLs and previous APOs(patient group),25 healthy pregnant females(HP group)and 35 healthy non-pregnant females(HNP group)from the Peking University Third Hospital,between October 2018 and March 2019,were enrolled in this study.Serum of HNP and serum of patients as well as HP in the first gestational trimester were collected.Enzyme-linked immunosorbent assay kits were used to measure serum BAFF and interferon-alpha(IFN-a)concentrations.Cytometric bead array analysis was used to measure serum concentrations of cytokines.The patient group was further divided into APOs and non-APOs(NAPOs)group,fetal loss and live birth group according to pregnancy outcomes.The Mann-Whitney U-test was used to assess significance between and within groups.Spearman rank-order was used to evaluate correlation coefficients between BAFF and related cytokines.Results:The serum BAFF level in HP group was significantly lower than HNP group(245.24[218.80,265.90]vs.326.94[267.31,414.80]pg/mL,Z=-3.966,P<0.001).The BAFF level was obviously elevated in patient group compared to that in HP group(307.77[219.86,415.65]vs.245.24[218.80,265.90]pg/mL,Z=-2.464,P=0.013).BAFF levels in APOs group tended to be higher than that in NAPOs group(416.52[307.07,511.12]vs.259.37[203.59,375.81]pg/mL,Z=-2.718,P=0.006).Compared to HP group,concentrations of IFN-a,interleukin(IL-6)and tumor necrosis factor were higher in patient group(33.37[18.85,48.12]vs.13.10[6.85,25.47]pg/mL,Z=-2.023,P=0.043;39.16[4.41,195.87]vs.3.37[2.92,3.90]pg/mL,Z=-3.650,P<0.001;8.23[2.27,64.46]vs.1.53[1.25,2.31]pg/mL,Z=-3.604,P<0.001,respectively).Serum BAFF levels had a positive correlation with the concentrations of both IL-6 and IL-10(IL-6:r=0.525,P=0.002;IL-10:r=0.438,P=0.012).Conclusions:Serum BAFF levels are increased in patients with positive aPLs and previous APOs as compared to healthy pregnant females and tend to be higher in individuals with current APOs.The BAFF levels have a positive correlation with serum IL-6 and IL-10.

River width and depth as key factors of diurnal activity energy expenditure allocation for wintering Spot-billed Ducks in the Xin'an River Basin

Rivers are important habitats for wintering waterbirds.However,they are easily influenced by natural and human activities.An important approach for waterbirds to adapt to habitats is adjusting the activity time and energy expenditure allocation of diurnal behavior.The compensatory foraging hypothesis predicts that increased energy expenditure leads to longer foraging time,which in turn increases food intake and helps maintain a constant energy balance.However,it is unclear whether human-disturbed habitats result in increased energy expenditure related to safety or foraging.In this study,the scan sample method was used to observe the diurnal behavior of the wintering Spot-billed Duck(Anas poecilorhyncha) in two rivers in the Xin’an River Basin from October 2021 to March 2022.The allocation of time and energy expenditure for activity in both normal and disturbed environments was calculated.The results showed that foraging accounted for the highest percentage of time and energy expenditure.Additionally,foraging decreased in the disturbed environment than that in the normal environment.Resting behavior showed the opposite trend,while other behaviors were similar in both environments.The total diurnal energy expenditure of ducks in the disturbed environment was greater than that in the normal environment,with decreased foraging and resting time percentage and increased behaviors related to immediate safety(swimming and alert) and comfort.These results oppose the compensatory foraging hypothesis in favor of increased security.The optimal diurnal energy expenditure model included river width and water depth,which had a positive relationship;an increase in either of these two factors resulted in an increase in energy expenditure.This study provides a better understanding of energy allocation strategies underlying the superficial time allocation of wintering waterbirds according to environmental conditions.Exploring these changes can help understand the maximum fitness of wintering waterbirds in response to nature and human influences.

Activity Data and Emission Factor for Forestry and Other Land Use Change Subsector to Enhance Carbon Market Policy and Action in Malawi

Activity data and emission factors are critical for estimating greenhouse gas emissions and devising effective climate change mitigation strategies. This study developed the activity data and emission factor in the Forestry and Other Land Use Change (FOLU) subsector in Malawi. The results indicate that “forestland to cropland,” and “wetland to cropland,” were the major land use changes from the year 2000 to the year 2022. The forestland steadily declined at a rate of 13,591 ha (0.5%) per annum. Similarly, grassland declined at the rate of 1651 ha (0.5%) per annum. On the other hand, cropland, wetland, and settlements steadily increased at the rate of 8228 ha (0.14%);5257 ha (0.17%);and 1941 ha (8.1%) per annum, respectively. Furthermore, the results indicate that the “grassland to forestland” changes were higher than the “forestland to grassland” changes, suggesting that forest regrowth was occurring. On the emission factor, the results interestingly indicate that there was a significant increase in carbon sequestration in the FOLU subsector from the year 2011 to 2022. Carbon sequestration increased annually by 13.66 ± 0.17 tCO2 e/ha/yr (4.6%), with an uncertainty of 2.44%. Therefore, it can be concluded that there is potential for a Carbon market in Malawi.

Effects of serum inflammatory factors,health index and disease activity scores on ankylosing spondylitis patients with sleep disorder

BACKGROUND Patients with ankylosing spondylitis(AS)frequently suffer from comorbid sleep disorders,exacerbating the burden of the disease and affecting their quality of life.AIM To investigate the clinical significance of serum inflammatory factors,health index and disease activity scores in patients with AS complicated by sleep disorders.METHODS A total of 106 AS patients with comorbid sleep disorders were included in the study.The patients were grouped into the desirable and undesirable prognosis groups in accordance with their clinical outcomes.The serum levels of inflammatory factors,including C-reactive protein,erythrocyte sedimentation rate,interleukin(IL)-6,tumour necrosis factor-αand IL-1β,were measured.Disease activity scores,such as the Bath AS functional index,Bath AS disease activity index,Bath AS metrology index and AS disease activity score,were assessed.The health index was obtained through the Short Form-36 questionnaire.RESULTS The study found significant associations amongst serum inflammatory factors,health index and disease activity scores in AS patients with comorbid sleep disorders.Positive correlations were found between serum inflammatory factors and disease activity scores,indicating the influence of heightened systemic inflammation on disease severity and functional impairment.Conversely,negative correlations were found between disease activity scores and health index parameters,highlighting the effect of disease activity on various aspects of healthrelated quality of life.Logistic regression analysis further confirmed the predictive value of these factors on patient outcomes,underscoring their potential utility in risk assessment and prognostication.CONCLUSION The findings demonstrate the intricate interplay amongst disease activity,systemic inflammation and patientreported health outcomes in AS patients complicated by sleep disorders.The results emphasise the need for comprehensive care strategies that address the diverse needs and challenges faced by these patients and underscore the potential relevance of serum inflammatory factors,health index and disease activity scores as prognostic markers in this patient population.

Lipopolysaccharide-induced cerebral inflammatory damage and the therapeutic effect of platelet activating factor receptor antoganist

Objective To investigate lipopolysaccharide （LPS） induced acute cerebral inflammatory damage and the therapeutic effect of ginkgolide B （BN52021）. Methods Thirty Sprague-Dawley rats were randomly divided into 3 groups （n = 10 for each group）： Control group, Model group and Treatment group （treated with BN52021）. LPS were injected into the fourth ventricle of rat to make a neuroinflammatory murine model. Morris water maze was used to detect the learning and memory ability of rats; changes of synapse number and subcellular ultrastructures were observed under a transmission electron microscope; OX-42 positive microglia in the brain was detected by immunohistochemical method. Results The average escape latency in the Treatment group were significantly shortened than that in the Model group; and the percentage of swimming distance traveled in platform quadrant accounting for total distance increased markedly. The rough endoplasmic reticulum and polyribosomes in the Treatment group were more than that in the Model group, but the number of synapses seemed to have no obvious change. The number of OX-42 positive microglia in the Treatment group decreased markedly than that in the Model group, and the grey density of OX-42-positive cells increased significantly. Conclusion LPS can induce inflammatory damages to the brain, but the damage could be antagonized by BN52021. Platelet activating factor receptor antagonist may offer an effective therapy for neurodegeneration diseases.

Elevated Platelet Activating Factor Level in Ischemia-Related Arrhythmia and Its Electrophysiological Effect on Myocardium

Objective The mechanism through which platelet activating factor （PAF） induces cardiac electrical activity and arrhythmia is not well understood and previous studies have suggested a potential involvement of ion channels in its action. The present study was aimed to clarify the role of PAF in fatal arrhythmias following acute myocardia infarction （AMI） and the underlying mechanism. Methods （1） Blood PAF levels were measured among 72 AMI patients at the time of diagnosis with AMI and 48 h later, and their electrocardiogram （ECG） was recorded continuously. （2） Ischemia simulation and surface electrocardiogram were conducted in 20 pigs and their PAF levels were measured. （3） PAF perfusion and standard microelectrode recording were performed on guinea pig papillarymuscles. Results In both humans and pigs, elevated PAF levels were detected in AMI and simulated ischemia, respectively, and even higher PAF levels were found when fatal arrhythmias occurred. In guinea pig myocardium, PAF induced a shortening of action potential duration at 90% level of repolarization （APD 90 ）under non-ischemic conditions and a more pronounced shortening under early simulated ischemic conditions. Conclusion AMI and ischemia are associated with increased PAF levels in humans and pigs, which are further raised when fatal arrhythmia follows. The effects of PAF on the myocardium may be mediated by multiple ion channels.

Significance of platelet activating factor receptor expression in pancreatic tissues of rats with severe acute pancreatitis and effects of BN52021

AIM:To investigate the dynamic changes and significance of platelet activating factor receptor (PAF-R) mRNA and protein in pancreatic tissues of rats with severe acute pancreatitis (SAP) and effects of BN52021 (Ginkgolide B). METHODS:Wistar male rats were randomly assigned to the negative control group (NC group),SAP model group (SAP group),and BN52051-remedy group (BN group),and each of the groups was divided into 6 subgroups at different time points after operation (1 h,2 h,3 h,6 h,12 h,and 24 h) (n=10 in each). PT-PCR and Western blot methods were used to detect PAF-RmRNA and protein expression in pancreatic tissues of rats respectively. Pathological examination of pancreatic tissues was performed and the serum amylase change was detected. RESULTS:Serum amylase and pathological results showed the that SAP model was successfully prepared,BN52021 was able to decrease serum amylase,and the pathological ratings in BN group at 3 h,6 h,and 12 h significantly decreased compared with those in the SAP group (8.85 ± 0.39 vs 5.95 ± 0.19,9.15 ± 0.55 vs 5.55 ± 0.36,10.10 ± 0.65 vs 6.72 ± 0.30,P < 0.05). The result of PAF-mRNA showed dynamic changes in SAP and BN groups,which increased gradually in early stage,reached a peak at 3 h (0.71 ± 0.14 vs 0.54 ± 0.14,0.69 ± 0.13 vs 0.59 ± 0.04,P < 0.05),and decreased gradually later. There were significant differences at each time point except 1 h and 2 h,when compared with those in the NC group (0.71 ± 0.14 or 0.69 ± 0.13 vs 0.47 ± 0.10,0.38 ± 0.08 or 0.59 ± 0.04 vs 0.47 ± 0.09,0.25 ± 0.07 or 0.29 ± 0.05 vs 0.46 ± 0.10,0.20 ± 0.06 or 0.20± 0.04 vs 0.43 ± 0.09,P < 0.05),whereas there was no significant difference between BN and SAP groups at each time point. The result of PAF-R protein showed that the change of PAF-R protein in the SAP group and the BN group was consistent with that of PAF-R mRNA. There were significant differences at each time point except 1 h,when compared with those in the NC group (0.90 ± 0.02 or 0.80 ± 0.05 vs 0.48 ± 0.02,1.69 ± 0.06 or 1.58 ± 0.02 vs 0.48 ± 0.03,1.12 ± 0.10 or 0.98 ± 0.03 vs 0.49 ± 0.09,1.04 ± 0.14 or 0.87 ± 0.02 vs 0.52 ± 0.08,0.97 ± 0.16 or 0.90 ± 0.05 vs 0.49 ± 0.10,P < 0.05),whereas there was no significant difference between the BN group and the SAP group. CONCLUSION:PAF-R plays an important role in occurrence and development of SAP. BN52021 exerts biological effects through competitively inhibiting the binding of increased both PAF and PAF-R expression rather than through decreasing PAF-R expression in pancreatic tissues.

Pathogenetic effects of platelet activating factor on enterogenic endotoxemia after burn

INTRODUCTION Previous clinical and experimental studies haveindicated that an early endotoxemia occurred after amajor burn.It is unlikely that burn wound sepsis isthe source of circulating endotoxin in less than 12hour after burn.Increasing evidence demonstratesthat the bacteria and endotoxin in thegastrointestinal tract can pass through the gutbarrier into blood circulation to form enterogenicendotoxemia following burn.However。

Platelet-activating factor in cirrhotic liver and hepatocellular carcinoma

AIM： Platelet-activating factor （PAF） is a pro-inflammatory and angiogenic lipid mediator. Here we aimed to investigate levels of PAF, lyso-PAF （the PAF precursor）, phospholipase A2 （PLA2, the enzymatic activity generating lyso-PAF）, acetylhydrolase activity （AHA, the PAF degrading enzyme） and PAF receptor （PAF-R） transcripts in cirrhotic liver and hepatocellular carcinoma （HCC）. METHODS： Twenty-nine patients with HCC were enrolled in this study. Cirrhosis was present in fourteen patients and seven had no liver disease. Tissue PAF levels were investigated by a platelet-aggregation assay. Lyso- PAF was assessed after its chemical acetylation into PAR AHA was determined by degradation of [^3H]-PAE PLA2 levels were assessed by EIA. PAF-R transcripts were investigated using RT-PCR. RESULTS： Elevated amounts of PAF and PAF-R transcripts 1 （leukocyte-type） were found in cirrhotic tissues as compared with non-cirrhotic ones. Higher amounts of PAF and PAF-R transcripts 1 and 2 （tissue-type） were found in HCC tissues as compared with non-tumor tissues. PLA2, lyso-PAF and AHA levels were not changed in cirrhotic tissues and HCC. CONCLUSION： While the role of PAF is currently unknown in liver physiology, this study suggests its potential involvement in the inflammatory network found in the cirrhotic liver and in the angiogenic response during HCC.

Synthesis of platelet-activating factor and its receptor expression in Kupffer cells in rat carbon tetrachloride-induced cirrhosis

AIM:To determine the platelet-activating factor (PAF) synthesis and its receptor expression in Kupffer cells in rat carbon tetrachloride-induced cirrhosis. METHODS:Kupffer cells, isolated from the livers of control and CCl4-induced cirrhotic rats, were placed in serum-free medium overnight. PAF saturation binding, ET-1 saturation and competition binding were assayed. ET-1 induced PAF synthesis, mRNA expression of PAF, preproendothelin-1, endothelin A (ETA) and endothelin B (ETB) receptors were also determined. RESULTS:A two-fold increase of PAF synthesis (1.42 ± 0.14 vs 0.66 ± 0.04 pg/μg DNA) and a 1.48-fold increase of membrane-bound PAF (1.02 ± 0.06 vs 0.69 ± 0.07 pg/μg DNA) were observed in activated Kupffer cells of cirrhotic rats. The application of ET-1 to Kupffer cells induced PAF synthesis in a concentration-dependent manner in both cirrhotic and normal rats via ETB receptor, but PAF synthesis in the activated Kupffer cells was more effective than that in the normal Kupffer cells. In activated Kupffer cells, PAF receptor expression and PAF binding capacity were markedly enhanced. Activated Kupffer cells raised the [125I]-ET-1 binding capacity, but changed neither the affinity of the receptors, nor the expression of ETA receptor. CONCLUSION:Kupffer cells in the course of CCl4-induced cirrhosis are the main source of increased PAF. ET-1 is involved endogenously in stimulating the PAF synthesis in activated Kupffer cells via ETB receptor by paracrine. ETA receptor did not appear in activated Kupffer cells, which may exacerbate the hepatic and extrahepatic complications of cirrhosis.

Effect of increased hepatic platelet activating factor and its receptor portal hypertension in CCl_4-induced liver cirrhosis

AIM： To evaluate the changes in hepatic platelet activating factor （PAF） and its receptors and their effect on portal pressure of cirrhotic rats induced by CCh. METHODS： A model of liver cirrhosis was replicated in rats by intra-peritoneal injection of CCh for 8 wk. We determined the effect of hepatic PAF and its receptor level on portal and arterial pressure by EIA, saturation binding and RT-PCR technique. RESULTS： Compared to control rats, cirrhotic rats had higher hepatic PAF levels and output as well as higher plasma PAF levels （P〈0.01, P〈0.01, P〈0.05, respectively）. Both hepatic PAF receptor mRNA levels and PAF binding were nearly 3-fold greater in cirrhotic rats （P〈0.01）. Portal injection of PAF （1 g/kg WT） increased the portal pressure by 22% and 33% in control and cirrhotic rats, respectively. In contrast, the arterial pressure was decreased in the both groups （54% in control rats and 42% in cirrhotic rats）. Injection of the PAF antagonist BN52021 （5 mg/kg WT） decreased the portal pressure by 16% in cirrhotic rats but had no effect in the control rats. CONCLUSION： The upregulation of the PAF system contributes to hepatic hemodynamic and metabolic abnormalities in drrhosis, and the increased release of PAF into the circulation has impacts on the systemic hemodynamics.

Effect of Buyang Huanwu Decoction on Platelet Activating Factor Content in Arterial Blood Preand Post-Arterial Thrombosis in Rats

To explore the effect of Buyang Huanwu Decoction (BHD) on platelet activating factor (PAF) content in arterial blood pre- and post-arterial thrombosis in rats, male Wistar rats were randomly divided into 3 groups, the medicine group treated with BHD, the control group with dexamethasone liquid, and the blank group with distilled water. Oral administration was given for 14 consecutive days, once daily. Model of arterial thrombosis was established in the animals 2 hours after final medication, the blood content of PAF, dry weight (DW) and occlusion time (OT) of thrombus, and dry weight of thrombus/body weight (TW/BW) ratio were observed. Results indicated that BHD could markedly lower the arterial blood content of PAF after thrombosis, increase the OT of thrombus, reduce the dry weight of thrombus and the TW/BW ratio (P

Sequential expression of cyclooxygenase-2, glutamate receptor-2, and platelet activating factor receptor in rat hippocampal neurons after fluid percussion injury

Traumatic brain injury causes gene expression changes in different brain regions. Occurrence and development of traumatic brain injury are closely related, involving expression of three factors, namely cyclooxygenase-2, glutamate receptor-2, and platelet activating factor receptor. However, little is known about the correlation of these three factors and brain neuronal injury. In this study, primary cultured rat hippocampal neurons were subjected to fluid percussion injury according to Scott’s method, with some modifications. RT-PCR and semi-quantitative immunocytochemical staining was used to measure the expression levels of cyclooxygenase-2, glutamate receptor-2, and platelet activating factor receptor. Our results found that cyclooxygenase-2 expression were firstly increased post-injury, and then decreased. Both mRNA and protein expression levels reached peaks at 8 and 12 hours post-injury, respectively. Similar sequential changes in glutamate receptor 2 were observed, with highest levels mRNA and protein expression at 8 and 12 hours post-injury respectively. On the contrary, the expressions of platelet activating factor receptor were firstly decreased post-injury, and then increased. Both mRNA and protein expression levels reached the lowest levels at 8 and 12 hours post-injury, respectively. Totally, our findings suggest that these three factors are involved in occurrence and development of hippocampal neuronal injury.

Hepatic stellate cells may be potential effectors of platelet activating factor induced portal hypertension

AIM: To determine platelet activating factor (PAF) receptor expression in cirrhotic hepatic stellate cells.METHODS: Hepatic stellate cells, isolated from the livers of control and CCl4-induced cirrhotic rats, were placed in serum-free medium after overnight culture. We determined the PAF receptor in hepatic stellate cells by saturation binding technique and semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR), and the effects of PAF and its antagonist BN52021 on prostaglandin E2 (PGE2) release by stellate cells.RESULTS: Scatchard analysis indicated the presence of PAF receptor with dissociation constant (Kd) of 4.66 nmol/L and maximum binding capacity (Bmax) of 24.65 fmol/μg in cirrhotic stellate cells. Compared with the control, the maximum PAF binding capacity increased significantly (Bmax: 24.65 ± 1.96 fmol/μg. DNA, R = 0.982 vs 5.74 ± 1.55 fmol/μg. DNA, R = 0.93; P < 0.01), whereas receptor affinity had no significant difference (Kd of 4.66 ± 0.33 nmol/L for the cirrhosis and 3.51 ± 0.26 nmol/L for the control; P > 0.05). Consistent with the receptor binding data, the mRNA expression of PAF receptor was increased significantly in cirrhotic stellate cells. PAF in a concentration-dependent manner induced PGE2 synthesis in cirrhotic hepatic stellate cells, but the effects were blocked significantly by BN52021.CONCLUSION: Cirrhosis sensitizes hepatic stellate cells to PAF by elevating its receptor level and hepatic stellate cells maybe potential effectors of PAF induced portal hypertension.

Creation of reversed phase high-performance liquid chromatographic technique to assay platelet-activating factor

Objective: To establish a new assay for platelet-activating factor (PAF), to compare it with bio-assay; and to discuss its significance in some elderly people diseases such as cerebral infarction and coronary heart disease. Methods: To measure PAF levels in 100 controls, 23 elderly patients with cerebral infarction and 65 cases with coronary heart disease by reversed phase high-performance liquid chromatographic technique (rHPLC). Results:rHPLC is more convenient, sensitive,specific, and less confusing, compared with bio-assay. The level of plasma PAF in patients with cerebral infarction was higher than that in the controls (P<0.01), and in patients with coronary heart disease. Conclusion: Detection of PAF with rHPLC is more reliable and more accurate. The new assay has important significance in PAF research.

Role of platelet-activating factor in reproduction:sperm function

Since its discovery nearly thirty years ago, platelet-activating factor has emerged as one of the more important lipidmediators known. Platelet-activating factor (PAF; 1-O-alkyl-2-O-acetyl-sn-glycero-3-phosphorylcholine) exists en-dogenously as a mixture of molecular species with structural variants of the alkyl moiety. PAF is a novel potent signal-ing phospholipid that has unique pleiotropic biological properties in addition to platelet activation. PAF also plays a sig-nificant role in reproduction. PAF content in squirrel monkey sperm is significantly higher during the breeding seasonthan the non-breeding season. PAF content in human sperm has a positive correlation with seminal parameters and preg-nancy outcomes. High-fertility boars have significantly more PAF in their sperm than low-fertility boars. The enzymes(lyso-PAF-acetyltransferase and PAF-acetylhydrolase) necessary for PAF activation and deactivation are present insperm. PAF-acetylhydrolase may act as a 'decapacitation factor'. Removal of this enzyme during capacitation maypromote PAF synthesis increasing motility and fertilization. PAF also plays a significant role in the fertilization process,enhancing the fertilization rates of oocytes. Enhanced embryo development has also been reported in oocytes fertilizedwith PAF-treated sperm. PAF antagonists inhibit sperm motility, acrosome reaction, and fertilization, thus suggestingthe presence of receptors for PAF. The PAF-receptor is present on sperm, with altered transcript levels and distributionpatterns on abnormal cells. Whereas the exact mechanism of PAF in sperm function and reproduction is uncertain, itsimportance in normal fertility is substantial. The reproductive significance of PAF activity in sperm and fertility plus therole of PAF in the establishment of pregnancy requires further study.

Prospective Cohort Investigation on Physical Activity of Osteoporosis Outcomes(PAOPO)in Jidong:Objectives,Study Design,and Baseline Characteristics

Objective The aim of this study was to investigate the prospective association between physical activity(PA),independently or in conjunction with other contributing factors,and osteoporosis(OP)outcomes.Methods The Physical Activity in Osteoporosis Outcomes(PAOPO)study was a community-based cohort investigation.A structured questionnaire was used to gather the participants’sociodemographic characteristics.Bone mineral density(BMD)measurements were performed to assess OP outcomes,and the relationship between BMD and OP was evaluated within this cohort.Results From 2013 to 2014,8,471 participants aged 18 years and older were recruited from Tangshan,China’s Jidong community.Based on their PA level,participants were categorized as inactive,moderately active,or very active.Men showed higher physical exercise levels than women across the activity groups.BMD was significantly higher in the very active group than in the moderately active and inactive groups.Individuals aged>50 years are at a higher risk of developing OP and osteopenia.Conclusion The PAOPO study offers promising insights into the relationship between PA and OP outcomes,encouraging the implementation of PA in preventing and managing OP.

Predictive active control of building structures using LQR and artificial intelligence

This study presents a neural network-based model for predicting linear quadratic regulator(LQR)weighting matrices for achieving a target response reduction.Based on the expected weighting matrices,the LQR algorithm is used to determine the various responses of the structure.The responses are determined by numerically analyzing the governing equation of motion using the state-space approach.For training a neural network,four input parameters are considered:the time history of the ground motion,the percentage reduction in lateral displacement,lateral velocity,and lateral acceleration,Output parameters are LQR weighting matrices.To study the effectiveness of an LQR-based neural network(LQRNN),the actual percentage reduction in the responses obtained from using LQRNN is compared with the target percentage reductions.Furthermore,to investigate the efficacy of an active control system using LQRNN,the controlled responses of a system are compared to the corresponding uncontrolled responses.The trained neural network effectively predicts weighting parameters that can provide a percentage reduction in displacement,velocity,and acceleration close to the target percentage reduction.Based on the simulation study,it can be concluded that significant response reductions are observed in the active-controlled system using LQRNN.Moreover,the LQRNN algorithm can replace conventional LQR control with the use of an active control system.

Roles of BN52021 in platelet-activating factor pathway in inflammatory MS1 cells

AIM: To determine the effects of BN52021 on platelet-activating factor receptor (PAFR) signaling molecules under lipopolysaccharide (LPS)-induced inflammatory conditions in MS1 cells. METHODS: MS1 cells (a mouse pancreatic islet endothelial cell line) were grown in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum, 2 mmol/L glutamine and 100 μg/mL penicillin/streptomycin in 5% CO 2 at 37 ℃. After growth to confluency in media, the cells were processed for subsequent studies. The MS1 cells received 0, 0.1, 1 and 10 μg/mL LPS in this experiment. The viability/prolifera-tion of the cells induced by LPS was observed using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide colorimetric assay. Apoptosis and necrosis of the cells under the inflammatory condition described previously were observed using Hoechst 33342-propidium iodide staining. Adenylate cyclase (AC), phospholipase A 2 (PLA 2 ), phospholipase Cβ (PLCβ), protein tyrosine kinase (PTK), G protein-coupled receptor kinases (GRK) and p38-mitogen-activated protein kinase (p38 MAPK) mRNA in the PAFR signaling pathway were measured by real-time polymerase chain reaction. The protein expression level of phosphorylated AC (p-AC), phosphorylated PLA 2 (p-PLA 2 ), phosphorylated PTK (p-PTK), phosphorylated p38 MAPK (p-p38 MAPK), PLCβ and GRK was measured using Western blotting analysis. RESULTS: The activity of MS1 cells incubated with dif- ferent concentrations of LPS for 6 h decreased significantly in the 1 μg/mL LPS group (0.49 ± 0.10 vs 0.67 ± 0.13, P < 0.05) and 10 μg/mL LPS group (0.44 ± 0.10 vs 0.67 ± 0.13, P < 0.001), but not in 0.1 μg/mL group. When the incubation time was extended to 12 h (0.33 ± 0.05, 0.32 ± 0.03 and 0.25 ± 0.03 vs 0.69 ± 0.01) and 24 h (0.31 ± 0.01, 0.29 ± 0.03 and 0.25 ± 0.01 vs 0.63 ± 0.01), MS1 cell activity decreased in all LPS concentration groups compared with the blank control (P < 0.001). BN52021 significantly improved the cell activity when its concentration reached 50 μmol/L compared with the group that received LPS treatment alone, which was consistent with the results obtained from fluorescence staining. The mRNAs levels of AC (4.02 ± 0.14 vs 1.00 ± 0.13), GRK (2.63 ± 0.03 vs 1.00 ± 0.12), p38 MAPK (3.87 ± 0.07 vs 1.00 ± 0.17), PLA 2 (3.31 ± 0.12 vs 1.00 ± 0.12), PLCβ (2.09 ± 0.08 vs 1.00 ± 0.06) and PTK (1.85 ± 0.07 vs 1.00 ± 0.11) were up-regulated after LPS stimulation as compared with the blank control (P < 0.05). The up- regulated mRNAs including AC (2.35 ± 0.13 vs 3.87 ± 0.08), GRK (1.17 ± 0.14 vs 2.65 ± 0.12), p38 MAPK (1.48 ± 0.18 vs 4.30 ± 0.07), PLCβ (1.69 ± 0.10 vs 2.41 ± 0.13) and PLA 2 (1.87 ± 0.11 vs 2.96 ± 0.08)were significantly suppressed by BN52021 except for that of PTK. The level of p-AC (1.11 ± 0.12 vs 0.65 ± 0.08), GRK (0.83 ± 0.07 vs 0.50 ± 0.03), PLCβ (0.83 ± 0.16 vs 0.50 ± 0.10) and p-p38 MAPK (0.74 ± 0.10 vs 0.38 ± 0.05) was up-regulated after LPS stimulation as compared with the blank control (P < 0.05). The up-regulated proteins, including p-AC (0.65 ± 0.15 vs 1.06 ± 0.14), GRK (0.47 ± 0.10 vs 0.80 ± 0.06), PLCβ (0.47 ± 0.04 vs 0.80 ± 0.19) and p-p38 MAPK (0.30 ± 0.10 vs 0.97 ± 0.05), was significantly suppressed by BN52021, but p-PLA 2 and p-PTK protein level were not suppressed. CONCLUSION: BN52021 could effectively inhibit LPS-induced inflammation by down-regulating the mRNA and protein levels of AC, GRK, p38 MAPK, PLA 2 and PLCβ in the PAFR signaling pathway.