Objective To study the value of MRI and proton magnetic resonance spectroscopy(H 1-MRS)for neonatal hypoxic-is-chemic encephalopathy(HIE).Methods Magnetic resonance imaging (MRI)and proton magnetic resonance spectroscopy (H 1-MRS)were performed in 30 cases of full-term neonates with HIE,and 10 infant control group without evidence of birth asphyxia. Cerebral MRI and H 1-MRS were performed within 1 5 days after birth.The results of H 1-MRS such as subwave crest values of me-tabolites in lesion areas were recorded.The data were analyzed statistically.Results (1)MRI showed abnormal fetures of HIE such as diffuse cerebral edema,loss of hyperintensity in the posterior limb of the internal capsule on T1 WI,gyrus sign,diffuse parenchy-mal hemorrhage,which could predict the severity of brain damage.(2)On H 1-MRS,the ratio of Lac/Cr in HIE group was much higer than that in control group,which was statistically significant (P <0.05).The ratio of Lac/Cr showed a rising trend with clini-cal grading of HIE.The ratio of NAA/Cr and NAA/Cho were lower in HIE group than that in control group (P <0.05),which showed a trend of gradually reduce with clinical grading of HIE.The difference between ratio of Glx-α/Cr in HIE group and control group was also significantly,the moderate-severe group was much higher than the mild group and control group.There was no sig-nificant difference in the ratio of Cho/Cr between the 4 groups.Conclusion The combination of MRI and H 1-MRS can objectively re-flect brain morphology and metabolic changes of HIE,and evaluate the severity of the brain injury,and provide an effective evidence for clinical diagnosis and treatment.

Objective To investigate the effect of cluster needling at scalp points plus gavage of modified Buyang Huanwu decoction on NSC differentiation in rats with ischemic stroke of qi deficiency and blood stasis type.Methods A model of qi deficiency and blood stasis syndrome was made using healthy rats. Dil dye was given for pre-labelling after the success of model making. The rats were randomized into groups A, B, C, D, E and F. Group F consisted of 4 rats and each of the other groups, of 12 rats. A rat model of ischemic stroke of qi deficiency and blood stasis type was made by MCAO at 48 hrs after pre-labelling. After the success of model making, group A was the model group, group B was treated by cluster needling at scalp points, group C was treated by an oral gavage of Buyang Huanwu decoction, group D was treated by an oral gavage of modified Buyang Huanwu decoction, group E was treated by cluster needling at scalp points plus an oral gavage of modified Buyang Huanwu decoction and group F was the sham operation group. The rats were sacrificed at various time points respectively and the materials were taken. By nerve function assessment, double immunofluorescence staining and laser scanning confocal microscopy, a comparative study was conducted to investigate the effects of different treatments on NSC differentiation.Results There was a statistically significant difference in the nerve function score between group B, C, D, E or F and group A after one week of treatment (P<0.01), between group B or E and group A after two weeks of treatment (P<0.05,P<0.01) and between group E and group C or D after one and two weeks of treatment (P<0.01,P<0.05). After one and two weeks of treatment, there were statistically significant differences in Brdu﹢/NeuN﹢ cell count, Brdu﹢/GFAP﹢ cell count, Dil﹢/Brdu﹢NeuN﹢ cell count and Dil﹢/Brdu﹢/GFAP﹢ cell count between group B, C, D or E and group A (P<0.01), between group D or E and group B or C (P<0.01) and between groups E and D (P<0.01).Conclusions Various treatments have a promoting effect on nerve stem cell differentiation in the rats. Of various treatments, cluster needling at scalp points plus gavage of modified Buyang Huanwu decoction has the best therapeutic effect.

OBJECTIVETo analyze the differential miRNA expression profile in the myocardium of rats with lipopolysaccharide (LPS)-induced endotoxemia and explore the role of miRNA in endotoxin-induced myocardial injury.

METHODSTwenty male SD rats received intraperitoneal injection of 10 mg/kg LPS (n=10) or an equivalent amount of saline solution (n=10). At 24 h after LPS injection, the rats were sacrificed to detect myocardial expressions of TLR4, TNF-α and IL-1β using real-time PCR and for observing myocardial ultrastructures under transmission electron microscopy. The differentially expressed miRNA in the myocardium were detected using a miRNA array, and the common differentially expressed miRNAs were selected for verifying their actual expressions using real-time PCR.

RESULTSTLR4, TNF-α and IL-1β were over-activated in the myocardium of LPS-treated rats, in which mitochondria swelling, structural damaged and cytoplasmic vacuoles were observed. In LPS-challenged rats, miR-194-3p, miR-344a-3p, miR-465-3p, miR-501-5p, miR-3596c, miR-185-3p, and miR-877 were found up-regulated significantly, whereas miR-208b-3p, miR-547-3p, miR-141-3p, miR-28-5p, and miR-3585-5p down-regulated in the myocardium.

CONCLUSIONSignificant differential expression of the miRNAs occurs in the myocardium of LPS-treated rats, suggesting their involvement in endotoxin-induced myocardial injury.

Animals ; Endotoxemia ; metabolism ; Injections, Intraperitoneal ; Interleukin-1beta ; metabolism ; Lipopolysaccharides ; Male ; MicroRNAs ; metabolism ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Toll-Like Receptor 4 ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; Up-Regulation

Objective To investigate the imaging features of necrotizing pneumonia(NP)caused by mycoplasma pneumoniae in children and to review the changes of serum CGreactive protein (CRP)and plasma DGdimer,which may provide an objective and effective help for clinicians.Methods 54 children with mycoplasma pneumoniae pneumonia (MPP)infection in our hospital were studied retrospectively,including 24 cases of NP and 30 cases of nonGNP (the control group).The dynamic changes of chest imaging,serum CRP and plasma DGdimer were compared between two groups.Results Chest imaging in NP group:All cases showed consolidation and necrosis,accompanied by pulmonary cavities,pleural effusion,bronchiectasia,and thickening of the bronchial wall.Decreased enhancement areas could be found in all 24 cases by enhanced CT.The review after treatment showed that the pulmonary consolidation was absorbted or narrowed,accompanied by cavities in lung,atelectasis,and pleura thickening.The imaging of non NP group showed mainly patches and spots shadow in lung, and only a little consolidation.After treatment,the lung lesions were basically absorbed.Only a few cases had fibrotic streaks and pleural thickening.The peak values of serum CRP in group NP and non NP group were respectively 91(33－266)mg/L,37(18－189) mg/L,and the duration of the abnormity were 1 9(1 1－3 8)d and 8(4－1 9)d.The peak values of plasma DGdimer in the 2 groups were respectively 2 3 78(1 9 84－5 908)ng/mL,1 7 6(1 2－41 9)ng/mL,and the duration of the abnormity were 24(13－64)d and 3(0－10)d. There were significant differences between the 2 groups with all parameters above.Conclusion The imaging characteristics of NP caused by MP in children include consolidation,necrosis (decreased enhancement areas in consolidation),pneumothorax ,cavities ,cystic degenerations, atelectasis,bronchial wall thickening,pleural effusion and pleura thickening.The course of NP was long,and the absorption was slow. The peaks value of serum CRP and plasma DGdimer in NP group were significantly higher than those in the non NP group,and the duration was longer than that in non NP group.Therefore,the clinicians should pay more attention to CR,DGdimer and chest imaging,which may help clinicians with the diagnosis and treatment.

Objective To analyze the differential miRNA expression profile in the myocardium of rats with lipopolysaccharide (LPS)-induced endotoxemia and explore the role of miRNA in endotoxin-induced myocardial injury. Methods Twenty male SD rats received intraperitoneal injection of 10 mg/kg LPS (n=10) or an equivalent amount of saline solution (n=10). At 24 h after LPS injection, the rats were sacrificed to detect myocardial expressions of TLR4, TNF-αand IL-1βusing real-time PCR and for observing myocardial ultrastructures under transmission electron microscopy. The differentially expressed miRNA in the myocardium were detected using a miRNA array, and the common differentially expressed miRNAs were selected for verifying their actual expressions using real-time PCR. Results TLR4, TNF-αand IL-1βwere over-activated in the myocardium of LPS-treated rats, in which mitochondria swelling, structural damaged and cytoplasmic vacuoles were observed. In LPS-challenged rats, miR-194-3p, miR-344a-3p, miR-465-3p, miR-501-5p, miR-3596c, miR-185-3p, and miR-877 were found up-regulated significantly, whereas miR-208b-3p, miR-547-3p, miR-141-3p, miR-28-5p, and miR-3585-5p down-regulated in the myocardium. Conclusion Significant differential expression of the miRNAs occurs in the myocardium of LPS-treated rats, suggesting their involvement in endotoxin-induced myocardial injury.

Objective To analyze the differential miRNA expression profile in the myocardium of rats with lipopolysaccharide (LPS)-induced endotoxemia and explore the role of miRNA in endotoxin-induced myocardial injury. Methods Twenty male SD rats received intraperitoneal injection of 10 mg/kg LPS (n=10) or an equivalent amount of saline solution (n=10). At 24 h after LPS injection, the rats were sacrificed to detect myocardial expressions of TLR4, TNF-αand IL-1βusing real-time PCR and for observing myocardial ultrastructures under transmission electron microscopy. The differentially expressed miRNA in the myocardium were detected using a miRNA array, and the common differentially expressed miRNAs were selected for verifying their actual expressions using real-time PCR. Results TLR4, TNF-αand IL-1βwere over-activated in the myocardium of LPS-treated rats, in which mitochondria swelling, structural damaged and cytoplasmic vacuoles were observed. In LPS-challenged rats, miR-194-3p, miR-344a-3p, miR-465-3p, miR-501-5p, miR-3596c, miR-185-3p, and miR-877 were found up-regulated significantly, whereas miR-208b-3p, miR-547-3p, miR-141-3p, miR-28-5p, and miR-3585-5p down-regulated in the myocardium. Conclusion Significant differential expression of the miRNAs occurs in the myocardium of LPS-treated rats, suggesting their involvement in endotoxin-induced myocardial injury.

Objective:To investigate the prognostic value of texture analysis of MRI diffusion weighted imaging (DWI) for neonatal hypoglycemic encephalopathy (HE).Methods:The clinical data and MRI data of 119 patients with neonatal HE admitted to Children′s Hospital of Nanjing Medical University from July 2013 to September 2020 were retrospectively analyzed. The children were followed up to 7—8 months and scored by Bayley scales of infant and toddler development. According to the overall development index, the children were divided into three groups: normal group (≥85, group A, n=42), mild developmental retardation group (70－84, group B, n=46) and developmental retardation group (≤69, group C, n= 31). The whole brain region (except sulcus and cisterna) was delineated as region of interest (ROI) by LIFEx 3.4 software in MRI apparent diffusion coefficient images. A total of 37 parameters were calculated automatically by the software, The clinical data, including gender, gestational age, age at MRI scan, birth weight, mode of delivery, history of asphyxia at birth, maternal preeclampsia or diabetes, minimum blood glucose, duration of hypoglycemia, neonatal behavioral neurological assessment (NBNA), presence or absence of polycythemia); the texture parameters, including histogram, volume, gray level co-occurrence matrix (GLCM), gray level run length matrix (GLRLM), neighborhood gray tone difference matrix (NGTDM), gray level size zone matrix (GLSZM), in the three groups were analyzed; and the diagnostic efficacy of clinical parameters and texture parameters was analyzed. Multivariate Logistic regression was used to analyze statistically significant clinical parameters and texture parameters, and receiver operating characteristic curve (ROC) was used to evaluate the prognostic efficacy of these parameter for neonatal HE. Results:There were no significant differences in gender, gestational age, age at MRI scan, delivery mode and blood glucose minimum among the three groups ( P>0.05). There were significant differences in birth weight [(3 150±130)g, (3 020±220)g, (2 880±140)g, F=-0.31, P=0.015], history of suffocation (10 cases, 18 cases, 20 cases, P=0.001), history of maternal diabetes or preeclampsia (14 cases, 29 cases, 21 cases, P=0.002), blood glucose duration [(5.0±0.2)d, (8.0±0.4)d, (14.0±1.7)d, F=-3.09, P=0.030] and NBNA scores (32.0±3.2, 28.0±2.6, 22.0±1.9, F=-4.21, P=0.010) among three groups. There were significant differences in kurtosis and entropy of histogram (2.57±1.12, 3.66±0.98, 4.23±0.37, F=3.54, P=0.010;5.89±1.09, 7.67±2.12, 8.92±1.62, F=-4.42, P=0.020); energy, contrast and dissimilarity of GLCM (0.48±0.01, 0.36±0.02, 0.23±0.01, F=-3.12, P=0.001;2 419±21, 3 354±31, 4 313±26, F=-4.16, P=0.020;126±14, 153±23, 344±43, F=-3.50, P<0.001); long run emphasis of GLRLM (0.78±0.15, 1.12±0.12, 1.76±0.31, F=-4.13, P=0.006), run length non-uniformity and run percentage (71.7±13.9, 96.6±10.7, 104.1±13.5, F=-0.98, P=0.001;0.91±0.05, 0.84±0.21, 0.72±0.17, F=2.97, P=0.010); coarseness and busyness of NGTDM [0.09±0.01, 0.13±0.03, 0.26±0.07, F=-1.95, P=0.003;0.16(0.04, 4.14), 0.32(0.05, 9.84), 0.45(0.15, 10.14), H=-3.24, P=0.030], short-zone emphasis and short-zone high gray length emphasis of GLSZM (4.74±0.45, 3.44±1.03, 1.88±0.67, F=-3.14, P=0.040; 278 963±239, 164 607±544, 111 653±618, F=-3.84, P=0.001) among three groups. Multivariate Logistic regression showed that duration of hypoglycemia, NBNA score, energy, kurtosis, run percentage and short zone effect were independent risk factors for poor prognosis of neonatal HE ( OR=7.43, 4.09, 1.10, 2.11, 1.36, 1.68, P=0.002, 0.027, 0.001, 0.006, 0.007, 0.010, respectively). ROC curve showed that for combined hypoglycemic duration, NBNA and texture parameters, the area under the curve (AUC) was the highest (AUC=0.94, P<0.001). Conclusion:Texture analysis of the MRI diffusion weighted imaging can predict the prognosis of neonatal hypoglycemic encephalopathy at an early stage, which has better prediction efficiency when combined with clinical features.

Objective:To explore the effects of acupuncture combined with Buyang Huanwu Decoction on intestinal flora in cerebral blood flow hypo perfusion model rats with carotid artery stenosis.Methods:Totally 40 rats were randomly divided into sham-operation group, model group, TCM treatment group and acupuncture and drug combination treatment group, with 10 rats in each group. Except the sham-operation group, the other groups were prepared cerebral ischemia model by needle control and thread embolism method. TCM treatment group received Buyang Huanwu Decoction 100 mg/kg for gavage, once a day, and the intervention lasted for 2 weeks. In the acupuncture and drug combination group, based on the TCM treatment group, Baihui and its left and right sides of 2 mm were selected for acupuncture, once a day, and continuous intervention was performed for 2 weeks. Neurological function evaluation and behavioral function score were performed 7 and 14 days after administration, respectively. 16S rRNA sequencing was used to comprehensively characterize the structure and composition of fecal microflora of rats in each group. Linear discriminant analysis Effect Size (LEfSe) was used to analyze the difference of intestinal bacteria among groups.Result:On the 7th and 14th day after administration, compared with the model group, the neurological function score in the TCM treatment group and the acupuncture and drug combination group decreased ( P<0.05), and the behavioral function score increased ( P<0.05). Compared with model group, the Shannon index of TCM treatment group and acupuncture and drug combination group increased ( P<0.05). The abundance of Firmicutes increased ( P<0.05), and the abundance of Bacteroidetes and Proteobacteria decreased ( P<0.05); the abundance of Clostridia increased ( P<0.05), and the abundance of Gammaproteobacteria decreased ( P<0.05). The abundance of Escherichia-Shigella and Bacteroides decreased ( P<0.05); the abundance of lactobacillus significantly increased ( P<0.05). Conclusion:Acupuncture combined with Buyang Huanwu Decoction can improve the symptoms of cerebral hypoperfusion model rats with carotid artery stenosis, and the mechanism may be to increase the abundance of probiotics.

ObjectiveTo explore the protective mechanism of paeoniflorin on mice with ulcerative colitis （UC） through the adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR） autophagy pathway. MethodUC mouse model was established by allowing mice freely drink 4% DSS， and 56 BALB/c male mice were randomly divided into model group， AMPK inhibitor group （20 mg·kg^-1）， paeoniflorin （50 mg·kg^-1） + inhibitor （20 mg·kg^-1） group， and high dose （50 mg·kg^-1）， medium dose （25 mg·kg^-1）， and low dose （12.5 mg·kg^-1） paeoniflorin groups. After seven days of drug intervention， the protective effect of paeoniflorin on mice with UC was determined by comparing the body weight， disease activity index （DAI） changes， and Hematoxylin-eosin （HE） staining results. Enzyme linked immunosorbent assay （ELISA） was used to detect the levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in the serum of mice in each group， and immunofluorescence was utilized to detect microtubule-associated protein 1 light chain 3 （LC3） content in the colon， AMPK， mTOR proteins， and their phosphorylated proteins including p-AMPK and p-mTOR in the colon tissue were detected by Western blot， and the mRNA expression levels of AMPK， mTOR， Beclin1， LC3， and p62 were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with the blank group， the model group showed a decrease in body mass， an increase in DAI score， and severe pathological damage to the colon. The levels of inflammatory factors including TNF-α and IL-6 increased in serum （P<0.01）， while the protein levels of LC3 and p-AMPK/AMPK were down-regulated in colon tissue， and those of p-mTOR/mTOR were up-regulated （P<0.01）. The mRNA expression levels of AMPK and LC3 were down-regulated， while the mRNA expression levels of mTOR and p62 were up-regulated （P<0.01）. Compared with the model group and the paeoniflorin + inhibitor group， the mice treated with paeoniflorin showed an increase in body mass， a decrease in DAI score， a reduction in pathological damage to colon tissue， and a reduction in the levels of inflammatory factors of TNF-α and IL-6 in serum （P<0.05）. The protein levels of LC3 and p-AMPK/AMPK in colon tissue were up-regulated， while the protein levels of p-mTOR/mTOR were down-regulated （P<0.01）. The mRNA expression levels of AMPK， Beclin1， and LC3 were up-regulated， while the mRNA expression of mTOR and p62 were down-regulated （P<0.01）. The colon tissue of the inhibitor group was severely damaged， and the trend of various indicators was completely opposite to that of the high dose paeoniflorin group. ConclusionPaeoniflorin can enhance autophagy and reduce inflammatory damage in mice with UC by activating the AMPK/mTOR signaling pathway and thus play a protective role.

ObjectiveTo explore the mechanism of Huanglian Jiedutang in inhibiting the pyroptosis mediated by NOD-like receptor protein 3 （NLRP3） inflammasomes and alleviating the acute liver injury （ALI） induced by lipopolysaccharide （LPS） in the mouse model of sepsis. MethodFifty-four male C57BL/6 mice were randomized into blank， model， low- （3.08 g·kg^-1）， medium- （6.15 g·kg^-1）， and high-dose （12.30 g·kg^-1） Huanglian Jiedutang， and positive control （dexamethasone） groups （n=9）. The mice were administrated with Huanglian Jiedutang at different doses by gavage for 7 days， and then LPS （15 mg·kg^-1） was injected intraperitoneally for the modeling of sepsis. In the positive control group， dexamethasone （0.05 g·kg^-1） was injected intraperitoneally 1.5 h after modeling， and the mouse sepsis score （MSS） was recorded 12 h after modeling. The mice were sacrificed for the collection of blood and liver tissue samples. The levels of alanine transaminase （ALT） and aspartate transaminase （AST） were measured by a biochemical analyzer. The levels of tumor necrosis factor （TNF）-α， interleukin （IL）-6， IL-1β， and IL-18 in the serum were measured by enzyme-linked immunosorbent assay kits. Hematoxylin-eosin staining was used to observe the pathological changes in the liver tissue. The content of NLRP3 was observed by the immunofluorescence assay. The expression of apoptosis-associated speck-like protein containing CARD （ASC） was detected by immunohistochemistry. The protein levels of NLRP3， ASC， Caspase-1， and gasdermin D （GSDMD） in the liver tissue were determined by Western blot. Real-time quantitative polymerase chain reaction（Real-time PCR） was employed to determine the mRNA levels of GSDMD， Caspase-1， IL-1β， and IL-18. ResultCompared with the blank group， the model group showed elevated levels of ALT and AST （P<0.01） and risen levels of inflammatory cytokines in the serum （P<0.01）. In addition， the modeling resulted in edema and necrosis in the liver， and up-regulated the protein levels of GSDMD， NLRP3， ASC， and Caspase-1 （P<0.01） and the mRNA levels of GSDMD， Caspase-1， IL-1β， and IL-18 （P<0.01）. Compared with the model group， the drug intervention groups showed reduced content of inflammatory cytokines （P<0.01）， alleviated pathological damage in the liver tissue， and down-regulated protein levels of GSDMD， NLRP3， ASC， and Caspase-1 （P<0.05，P<0.01） and mRNA levels of GSDMD， Caspase-1， IL-1β， and IL-18 （P<0.05，P<0.01） in the liver tissue. ConclusionHuanglian Jiedutang can inhibit pyroptosis and reduce inflammation by inhibiting the activation of NLRP3 inflammasomes， thus demonstrating a therapeutic effect on acute liver injury in the mouse model of sepsis induced by LPS.