Airway mucus hypersecretion is one of the pathological changes in children with community acquired pneumonia（CAP），and affects the severity，progression and prognosis of CAP.Diagnosis of airway mucus hypersecretion currently relies on fiberoptic bronchoscopy. To predict the risk of airway mucus hypersecretion and to take early action to avoid serious consequences such as plastic bronchitis and hypoxia and respiratory failure，the article summarizes the relationship between age，clinical characteristics and laboratory indices and the formation of airway mucus hypersecretion in children with CAP. Among them，age，pathogenic species，fever peak and fever range，neutrophil ratio，C-reactive protein，lactate dehydrogenase，D-dimer，serum 25（OH）D，and some interleukins，interferons，and acquired immune indicators have an early warning effect on the development of airway mucus hypersecretion in children with CAP.

AIM:To investigate the mechanism of Piezo1 channel activation promoting the increase in intra-cellular Ca2+ concentration([Ca2+]i)of rat coronary artery smooth muscle cells(CASMCs).METHODS:The primary CASMCs of SD rats were cultured,and the expression and subcellular localization of Piezo1 in the cells were observed by immunofluorescence staining.The Piezo1 and stromal interaction molecule 1(STIM1)in CASMCs were knocked down by siRNA transfection,and the expression levels of the proteins were detected by Western blot.Utilizing laser confocal mi-croscopy,the change of[Ca2+]i in CASMCs was detected by Fluo-4 AM fluorescent probes.RESULTS:It was confirmed by immunofluorescence staining that the expression of Piezo1 existed in primary rat CASMCs.Immunofluorescence staining also showed that Piezo1 was co-located with sarco-/endoplasmic reticulum Ca2+-ATPase 2(SERCA2),mitochondrial outer membrane protein TOM20 and nuclear membrane protein lamin B1.Western blot results showed that the protein expres-sion levels of STIM1 and Piezo1 were significantly down-regulated by siRNA transfection(P<0.05).Compared with con-trol group,Yoda1,the agonist of Piezo1,could increase the extracellular Ca2+ influx of CASMCs(P<0.01).However,the Ca2+ influx mediated by Yoda1 was not affected by the inhibition of L-type calcium channels.Treatment with Yoda1 in-creased the intracellular Ca2+ release of CASMCs(P<0.01).However,inhibition of calcium channels on endoplasmic re-ticulum,ryanodine receptor and inositol 1,4,5-triphosphate receptor,did not affect intracellular Ca2+ release mediated by Yoda1.After the Ca2+ in endoplasmic reticulum was emptied using thapsigargin(TG),Yoda1 also mediated the Ca2+ re-lease of other organelles in CASMCs(P<0.01).After inhibition of L-type calcium channels,treatment with store-operated calcium channel(SOCC)inhibitor BTP2 or knockdown of STIM1 led to the decrease in extracellular Ca2+ influx of CASMCs mediated by Yoda1(P<0.01).Treatment with TG increased the release of Ca2+ from the endoplasmic reticulum of CASMCs after knockdown of Piezo1(P<0.05),but the extracellular Ca2+ influx mediated by TG was not affected.After inhibition of L-type calcium channels and SOCC,knockdown of Piezo1 led to the decreases in intracellular Ca2+ release and extracellular Ca2+ influx induced by Yoda1(P<0.01).CONCLUSION:The Piezo1 agonist orchestrates the influx of extracellular Ca2+ by activating Piezo1 channels on the cell membrane and inducing the indirect activation of SOCC.More-over,it facilitates the release of Ca2+ from organelles.Consequently,these pathways synergistically elevate the[Ca2+]i of rat CASMCs.

AIM:To investigate the influence of fat mass and obesity-associated(Fto)gene on the aberrant contraction of aortic smooth muscle in diabetes mellitus(DM)mice,and to explore the mechanism of Fto gene underlying the calcium regulation.METHODS:Smooth muscle-specific Fto gene knockout(FtoSMKO)mice were generated using Cre-loxP technology.The experiment involved 3 groups of mice:wild-type(WT)group,DM model group and FtoSMKO-DM group,with 15 mice in each group.In DM group and FtoSMKO-DM group,type 1 DM was induced by intraperitoneal injec-tion of streptozotocin.The mice in WT group were injected with equal volume of citric acid-sodium citrate buffer solution.The influences of different drugs on the contraction responses of aortic smooth muscle in mice were analyzed using a multi-myograph system.The expression level of FTO protein in the aortic tissues was detected by Western blot.RESULTS:(1)Compared with WT mice,the expression levels of FTO protein in the aortic tissues of DM mice were significantly in-creased(P＜0.01).(2)The expression level of FTO protein in smooth muscle was significantly decreased after knockout of Fto gene(P＜0.01).Compared with WT group,the mice in DM group exhibited a significant decrease in body weight and a marked increase in fasting blood glucose level(P＜0.05).There were no noticeable differences in body weight or fasting blood glucose level between FtoSMKO-DM group and DM group(P＞0.05).(3)The contraction responses of aortic smooth muscle in DM group were substantially increased by phenylephrine compared with WT group.Specifically,vaso-constriction responses mediated by non-L-type calcium channels and store-operated calcium channels(SOCC)were signifi-cantly enhanced in DM group.In addition,the responses mediated by inositol 1,4,5-trisphosphate receptors(IP3R),which facilitate calcium release from the sarcoplasmic reticulum,were significantly enhanced.However,the responses mediated by caffeine-activated ryanodine receptors(RyR),which also facilitate calcium release from the sarcoplasmic re-ticulum,were significantly inhibited(P＜0.05).(4)Compared with DM group,the phenylephrine-induced contraction re-sponses of aortic smooth muscle in FtoSMKO-DM group were greatly weakened(P＜0.05).In particular,the vasoconstriction responses mediated by non-L-type calcium channels and SOCC in FtoSMKO-DM group were greatly suppressed(P＜0.05),while those mediated by caffeine-activated RyR were dramatically boosted(P＜0.05).However,IP3R-mediated responses were not affected(P＞0.05).CONCLUSION:Smooth muscle-specific Fto gene knockout suppresses contractile hyperre-sponsiveness in the aortic smooth muscle of DM mice,which may be attributed to involvement of FTO protein in calcium regulation in the vascular smooth muscle.

AIM:The objective of this study is to examine the expression of profilin 1(PFN1)in mice with di-abetic nephropathy and determine its association with immune cell infiltration.METHODS:This study presents an analy-sis of PFN1 expression and immune cell infiltration in patients with diabetic nephropathy,utilizing transcriptome expres-sion data from kidney tissue microarray.Additionally,the findings were validated in a diabetic nephropathy mouse model.Sixteen C57BL/6 mice were randomly assigned into two groups,namely the normal group and the model group,in an equal manner.The model group underwent the establishment of the diabetic nephropathy model through intraperitoneal injection of streptozotocin.Subsequently,the expression levels of CD11b,F4/80,CC chemokine receptor 4(CCR4),interleukin-1 receptor type I(IL-1R1),B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax)and caspase-3 in kidney tissue were assessed upon successful establishment of the diabetic nephropathy model.Furthermore,the overexpression of PFN1 was observed in a cellular model of diabetic nephropathy,and the protein expression levels of monocyte chemotactic pro-tein-1(MCP-1)and caspase-3 were assessed.RESULTS:The expression of PFN1 was found to be significantly in-creased in the GSE30122 dataset of transcriptome expression in kidney tissues affected by diabetic nephropathy(P＜0.01).This increase in PFN1 expression was found to be correlated with the presence of macrophages and T cells.Fur-thermore,the renal tissue of the diabetic nephropathy model group exhibited significant pathological changes.In this mod-el group,the expression levels of PFN1,CD11b,F4/80,CCR4,IL-1R1,Bax,Bcl-2,and caspase-3 were all significant-ly increased(P＜0.01).Overexpression of PFN1 could enhance the expression of MCP-1 and caspase-3 proteins.CON-CLUSION:Macrophages and Th17 cells were identified within the renal tissue of mice with diabetic nephropathy,con-comitant with an up-regulation in the expression of PFN1.This up-regulation was observed to facilitate the induction of apoptosis in the context of diabetic nephropathy.

There is an accumulating body of evidence implicating the muscarinic acetylcholine receptor 4 (M₄) in schizophrenia and dementia with Lewy bodies, however, a clinically validated M₄ positron emission tomography (PET) radioligand is currently lacking. As such, the aim of this study was to develop a suitable M₄ PET ligand that allows the non-invasive visualization of M₄ in the brain. Structure-activity relationship studies of pyrazol-4-yl-pyridine derivates led to the discovery of target compound 12 - a subtype-selective positive allosteric modulator (PAM). The radiofluorinated analogue, [¹⁸F] 12, was synthesized in 28 ± 10% radiochemical yield, >37 GBq/μmol and an excellent radiochemical purity >99%. Initial in vitro autoradiograms on rodent brain sections were performed in the absence of carbachol and showed moderate specificity as well as a low selectivity of [¹⁸F] 12 for the M₄-rich striatum. However, in the presence of carbachol, a significant increase in tracer binding was observed in the rat striatum, which was reduced by >60% under blocking conditions, thus indicating that orthosteric ligand interaction is required for efficient binding of [¹⁸F] 12 to the allosteric site. Remarkably, however, the presence of carbachol was not required for high specific binding in the non-human primate (NHP) and human striatum, and did not further improve the specificity and selectivity of [¹⁸F] 12 in higher species. These results pointed towards significant species-differences and paved the way for a preliminary PET study in NHP, where peak brain uptake of [¹⁸F] 12 was found in the putamen and temporal cortex. In conclusion, we report on the identification and preclinical development of the first radiofluorinated M₄ PET radioligand with promising attributes. The availability of a clinically validated M₄ PET radioligand harbors potential to facilitate drug development and provide a useful diagnostic tool for non-invasive imaging.

Objective: To analyze of epidemiological and etiological characteristics of 14 norovirus clusters or outbreak in Nan an District, for comprehensive prevention and control measures for norovirus infections in the region. Methods: Data were collected from the emergency public health event management information system of China Disease Prevention and Control Information System, and were analyzed by using descriptive epidemiological method. Results: In 2018, 14 cases of norovirus infection clusters and outbreaks were reported in Nan an District, accounting for 63.64% of the total number of incidents in the region. A total of 268 cases were reported, with an average incidence of 2.19%; the outbreak occurred mainly in November(n=6); kindergartens reported the most outbreak(n=7), followed by primary schools(n=5); the median duration of the outbreak was 2.80 days; and 14 outbreaks were caused by the GII-type genome of norovirus, with the main transmission routes being human-to-hnuman transmission. Conclusion The prevalence of norovirus outbreaks tends to be higher in schools, it is necessary to strengthen the monitoring of vomiting and diarrhea symptoms in collective units such as schools, and efforts should be promoted for implementation of all levels of prevention and control measures in school.

Objective To investigate the clinical efficacy of dexmedetomidine (DEX) combined with pentazocine intravenous anesthesia in percutaneous kyphoplasty (PKP) for osteoporotic vertebral compression fracture (OVCF) in the elderly.Methods A retrospective case-control study was performed to analyze 63 elderly OVCF patients treated with PKP and admitted to Tianjin Hospital from June 2018 to December 2018.There were 20 males and 43 females,aged 65-86 years [(74.7 ± 1.1)years].There were 15 patients with thoracic compression fractures and 48 with lumbar compression fractures,in whom the vertebral height loss was ＜ 30％ without posterior ligament complex damage.Nineteen patients received local anesthesia with lidocaine (Group A),21 patients received intravenous anesthesia with pentazocin and propofol (1 mg/kg) (Group B),and 23 patients received intravenous anesthesia with pentazocin and DEX (0.5 μg/kg) (Group C).Heart rate,systolic blood pressure,respiratory rate and blood oxygen saturation (SPO2) were recorded at 5 time points:at rest in the operating room (T0),after intravenous injection of pentazocine (T1),when the balloon dilated (T2),after the injection of cement (T3),and blinking after being called or at the end of the operation (T4).The levels of plasma cortisol were recorded before and at T3 in three groups.Visual analogue score (VAS) at T4 was recorded.The operation time,patient satisfaction and incidence of adverse reactions were recorded.The wake-up time and orientation recovery time of groups B and C were recorded.Results The heart rate,mean systolic blood pressure,respiratory rate in Group A at T2,T3 and T4 were higher than those in Groups B and C (P ＜ 0.05),but there was no significant difference between Groups B and C (P ＞ 0.05).There was no significant difference in SPO2 at only time among the groups (P ＞ 0.05).No significant differences were found in cortisol between the three groups before surgery.The cortisol level of Group A at T3 was higher than those of Groups B and C with significant difference (P ＜ 0.05),but no significant difference was found between Groups B and C (P ＞0.05).The VAS in Group A was significantly higher than those in groups B and C (P ＜ 0.01).The operative time in Group A was longer than those in Groups B and C (P ＜ 0.05),but no significant difference was found between Groups B and C (P ＞ 0.05).There was no significant difference in patient satisfaction between Groups B and C,and both of them were higher than Group A (P ＜0.01).There was no significant difference in the incidence of adverse reactions between the three groups (P ＞ 0.05).The wake-up time and orientation recovery time of Group C were shorter than those of Group B (P ＜ 0.01).Conclusion For elderly OVCF patients,pentazocin combined with propofol or DEX can be applied in PKP,which has satisfactory analgesic effect,slight effect on respiratory and circulatory,less adverse reactions,and good patient feedback.DEX has more advantages of awakening and orientation recovery and is worthy of clinical application.

Endoplasmic reticulum (ER) stress is mediated by disturbance of Ca²⁺ homeostasis. The store-operated calcium (SOC) channel is the primary Ca²⁺ channel in non-excitable cells, but its participation in agent-induced ER stress is not clear. In this study, the effects of tunicamycin on Ca²⁺ influx in human umbilical vein endothelial cells (HUVECs) were observed with the fluorescent probe Fluo-4 AM. The effect of tunicamycin on the expression of the unfolded protein response (UPR)-related proteins BiP and CHOP was assayed by western blotting with or without inhibition of Orai1. Tunicamycin induced endothelial dysfunction by activating ER stress. Orai1 expression and the influx of extracellular Ca²⁺ in HUVECs were both upregulated during ER stress. The SOC channel inhibitor SKF96365 reversed tunicamycin-induced endothelial cell dysfunction by inhibiting ER stress. Regulation of tunicamycin-induced ER stress by Orai1 indicates that modification of Orai1 activity may have therapeutic value for conditions with ER stress-induced endothelial dysfunction.
Blotting, Western ; Calcium ; Endoplasmic Reticulum ; Endoplasmic Reticulum Stress ; Endothelial Cells ; Homeostasis ; Human Umbilical Vein Endothelial Cells ; Tunicamycin ; Unfolded Protein Response

Objective To study the effects of mineral oil covered M2 culture medium droplet culture, M16 droplet culture, and dimethyl sulfoxide (DMSO) on the morphology and survival rates of mouse oocytes during the release from diplotene arrest. Methods Oocytes were randomly divided into 3 groups and individually cultured for 4 h in M2 covered with mineral oil, M16 covered with mineral oil, and/or M16 only to cause germinal vesicle breakdown (GVBD). The morphological changes and survival rates of oocytes in each group were observed under the microscope. Oocytes were randomly divided into 3 groups and cultured in the medium with 0％, 1％, and 2％ DMSO. The effect of DMSO on oocytes was also observed during the release from diplotene arrest. Results The survival rates of oocytes in M2 covered with mineral oil were higher than those in M16 (P ＜ 0.05). There was no statistical difference with respect to release of mouse oocytes from diplotene arrest between the oocytes in M2 covered with mineral oil and oocytes in M16. The shape of oocytes in M2 with mineral oil was better than that of oocytes in M16. The effect of DMSO on the survival rate of oocytes was similar in the medium with 0％, 1％ and 2％DMSO. But the effect of 2％ DMSO on the release of oocytes was statistically significant (P ＜ 0.01). Conclusion During the release of mouse oocytes from diplotene arrest, oocytes in M2 covered with mineral oil have much better morphology and higher survival rate than those in M16. DMSO (0％, 1％ and 2％) has no effect on the survival rate of oocytes. However, 2％ DMSO is more effective in promoting the release of mouse oocytes from diplotene arrest.

AIM: To investigate the primary culture method for coronary artery smooth muscle cells (CASMCs), and to establish the endoplasmic reticulum stress ( ERS) model in CASMCs of SD rats.METHODS:CASMCs were cultured by tissue explant method .The morphological characteristics were observed under optical micro-scope.The marker proteins of CASMCs , including α-SMA and SM-MHC, were identified by immunofluorescence tech-nique.The protein expression levels of BiP and CHOP , the marker molecules of ERS, were determined by Western blot . RESULTS:The spindle-shaped CASMCs climbed out from the edge of coronary artery tissues after 6 d, and formed the typical hill and valleygrowth pattern of CASMCs at 9~10 d.The result of immunofluorescence technique showed that α-SMA and SM-MHC were positively expressed .The results of Western blot showed that the protein expression of BiP and CHOP in TG ( 1 and 2 μmol/L ) treatment groups was increased compared with control group .Compared with control group, the protein expression of BiP and CHOP was significantly increased after 1 μmol/L TG treatment for 24 and 48 h. CONCLUSION:CASMCs can be successfully cultured by tissue explant method .ERS model of CASMCs was established by 1 μmol/L TG treatment for 24 h.