OBJECTIVE To investigate ameliorative effects and mechanisms of two probiotics （Bacillus subtilis， Lactobacillus acidophilus） combined with Aurantii Fructus Immaturus （AFI） on functional dyspepsia （FD） in rats. METHODS Rats were randomly divided into blank group， model group， positive control group （domperidone group， 2.7 mg/kg）， AFI group （9 g/kg）， L. acidophilus group （5×107 cfu/kg）， B. subtilis group （5×107 cfu/kg）， L. acidophilus+ AFI group （L. acidophilus 5×107 cfu/kg+ AFI 9 g/kg）， and B. subtilis+AFI group （B. subtilis 5×107 cfu/kg+AFI 9 g/kg）， with 8 rats in each group. Except for the blank group， FD model was established by tail-clamping stimulation+hunger and satiety disorder+swimming exhaustion in other groups. After modeling， each group was given the corresponding drug/probiotic suspensions/physiological saline intragastrically， once a day， for 14 consecutive days. After the last medication， gastric emptying rate and the rate of propulsion of the small intestine in rats were measured； the levels of brain-gut peptide-related indicators [gastrin （GAS）， substance P （SP）， vasoactive intestinal peptide （VIP）， somatostatin （SS） and cholecystokinin （CCK）] in the serum of rats were measured. The pathological morphology of the gastric antrum tissue and duodenal tissue was observed. Cecal contents from the rats were collected for gut microbiota sequencing analysis. The protein expression levels of tyrosine kinase receptor c-Kit and stem cell factor （SCF） in the gastric antrum tissue， as well as Toll-like receptor 4 （TLR4）， myeloid differentiation factor 88 （MyD88）， and nuclear factor κB （NF-κB） in the duodenal tissue of the rats were detected. RESULTS Compared with the blank group， model group showed significantly lower gastric emptying rate， small intestinal propulsion rate， serum levels of GAS and SP， relative abundance of Firmicutes， Ace， Chao and Sobs indexes of the gut microbiota， and protein levels of SCF and c-Kit in gastric antrum （P＜0.05）， while serum levels of VIP， SS and CCK， relative abundance of Bacteroidetes， as well as protein expressions of TLR4， MyD88， and NF-κB， were significantly higher （P＜0.05）. The histological structure JZYC23S53） of the gastric antrum tissue appeared basically normal； however， abnormalities were observed in the duodenal structure， with a significant infiltration of inflammatory cells visible. Compared with the model group， all treatment groups significantly modulated most of the above indexes （P＜0.05）. The histological structure of the gastric antrum tissue was normal. Except for the B. subtilis group and the B. subtilis+AFI group， the pathological states of the duodenum in the remaining rats gradually recovered. Compared with each single drug group， most of above indexes in rats from each combination group showed further improvement （P＜0.05）. CONCLUSIONS The combination of AFI with two probiotics can improve gastrointestinal motility in FD rats， and the effect is superior to that of using the drugs alone. The specific underlying mechanisms may be related to the activation of the SCF/c-Kit signaling pathway and the inhibition of the TLR4/MyD88/NF-κB signaling pathway.

Objective:To investigate the alterations in brain dynamic functional network connectivity (dFNC) and their significance in Parkinson's disease (PD) patients with postural instability/gait difficulty (PIGD).Methods:Ninety PD patients admitted to Department of Neurology, First Affiliated Hospital of Nanjing Medical University from May 2016 to August 2019 were recruited, and 54 healthy controls matched with gender and age were chosen; their clinical data and resting-state functional MRI (rs-fMRI) data were collected. PD patients were divided into PD with PIGD (PD-PIGD) group ( n=49) and PD without PIGD (PD-non-PIGD) group ( n=41) according to Unified Parkinson's Disease Rating Scale (UPDRS) scores. Independent component analysis (ICA), sliding window method and k-means clustering were used to analyze the dFNC and compare among groups. Correlations of dFNC alterations with clinical scales were verified by partial correlation analysis. Results:Four repeated recurring functional connectivity states were identified, and PD-PIGD patients had high frequency in state 3 (44%) and state 2 (23%) of the low dFNC. In terms of dFNC time attributes, PD-PIGD patients had longer mean dwell time in state 3 than PD-non-PIGD patients and had lower number of transitions in state 3 than PD-non-PIGD patients and healthy controls, with significant differences ( P<0.05); PD-PIGD patients had significantly higher fractional windows and statistically longer mean dwell time in state 2 than healthy controls ( P<0.05). In terms of dFNC strengths, compared with healthy controls, PD-PIGD patients showed significantly decreased functional connectivity within default mode network (DMN, between medial superior frontal gyrus and precuneus) and auditory network (AN, between superior temporal gyrus and middle temporal gyrus), but significantly increased functional connectivity between sensorimotor network (SMN, supplementary motor area) and DMN (precuneus) in state 2 ( P<0.05, false discovery rate [FDR]-corrected). Partial correlation analysis indicated positive correlation between mean dwell time in state 3 and PIGD scores in PD-PIGD patients ( r=0.450, P=0.039). Conclusion:PD-PIGD patients exhibit specific dFNC, mainly characterized by low connectivity of the brain functional network and prolonged dwell time; local functional network domains often separate between DMN, AN and SMN networks and within the networks.

With the dramatically increasing detection rate of ground-glass nodules (GGN), exact understanding and treatment strategy of them has become the hottest issue currently. More and more studies have begun to explore the underlying mechanisms of their indolent characteristics and favorable prognosis from the perspectives of molecular evolution and immune microenvironment. GGN has different dominating gene mutations at different evolutional stages. The pure GGN has a lower tumor mutation burden and genomic instability, while a gradually evolutionary feature of genomic mutation along with the pathological progression can be observed. GGN has less infiltration of immune cells, and they are under the pressure of immune surveillance with weakened immune escape. With the increase of solid components, an inhibitory immune microenvironment is gradually established and immune escape is gradually enhanced, leading to rapid tumor growth. Further exploration of the molecular characteristics of GGN will help to more precisely distinguish these highly heterogeneous lesions, which could be helpful to make personalized treatment plans.

With the dramatically increasing detection rate of ground-glass nodules (GGN), exact understanding and treatment strategy of them has become the hottest issue currently. More and more studies have begun to explore the underlying mechanisms of their indolent characteristics and favorable prognosis from the perspectives of molecular evolution and immune microenvironment. GGN has different dominating gene mutations at different evolutional stages. The pure GGN has a lower tumor mutation burden and genomic instability, while a gradually evolutionary feature of genomic mutation along with the pathological progression can be observed. GGN has less infiltration of immune cells, and they are under the pressure of immune surveillance with weakened immune escape. With the increase of solid components, an inhibitory immune microenvironment is gradually established and immune escape is gradually enhanced, leading to rapid tumor growth. Further exploration of the molecular characteristics of GGN will help to more precisely distinguish these highly heterogeneous lesions, which could be helpful to make personalized treatment plans.

Objective    To emphasize the important role of video-assisted thoracoscopic surgery (VATS) in treatment of mediastinal bronchogenic cysts (MBCs). Methods    We retrospectively reviewed the clinical data of 112 patients (53 males and 59 females) of mediastinal bronchogenic cysts who underwent VATS in our institution between April 2001 and Aprial 2016. Median age was 4–75 (45.6±15.0) years. All patients underwent chest CT preoperatively. The patients were divided into two groups: an anterior mediastinum group, 47 patients; a middle and posterior mediastinum group, 65 patients including 35 patients in the middle mediastinum, 30 patients in the posterior mediastinum. The average diameter was 0.5–22.0 (3.50±2.33) cm. The average CT attenuation was 0–67 (35.5±15.3) Hu on unenhanced CT. We began each operation with the VATS technique. Results    The CT diagnostic accuracy for group middle and posterior mediastinum with CT value≤20 Hu was higher than others (61.5% vs. 13.1%, χ2=17.675, P<0.001). A total of 111 patients underwent VATS, only one patient converted to open thoracotomy. Cyst resection and thymectomy were conducted in 45 patients, cyst resection and extended thymectomy were conducted in 2 patients in the anterior mediastinum group. Simply cyst resection were performed in the middle and posterior mediastinum group (n=65). The average operative time was 40–360 (104.5±43.1) min. The average intraoperative blood loss was 5–600 (57.9±88.9) mL. The intraoperative complication rate was 3.6% and the incomplete resection rate was 6.3%. The main reason for these was severe adhesion between the cyst and mediastinal structure. No serious postoperative complication was found. Follow-up was done in 99 patients, and the mean follow-up time was 42 (12–191) months. There was no local recurrence. Conclusion    VATS resection of MBCs is a safe and efficacious procedure, and minimally invasive and  surgical resection should be performed as early as possible for MBCs.

Objective To study the relationship between plasma neurodegenerative protein level and non-motor symptoms(NMS)in PD patients.Methods Eighty-four PD patients served as a PD group and 54age-matched persons undergoing physical examination served as a control group. The NMS of PD patients were assessed according to the HAMD scale.The plasma levels of tau,p-tau181,Aβ-42andα-syn were measured by ELISA and analyzed by Spearman correlation analysis and binary logistic regression analysis respectively.Results The FSS score and plasmaα-syn level were significantly higher while the plasma Aβ-42level was significantly lower in PD group than in control group(3.22±1.68 vs 1.89±1.16,P=0.000;320.00±64.91ng/L vs 277.78±52.75ng/L,P=0.000;267.61±77.75ng/L vs 321.80±49.41ng/L,P=0.001).No significant difference was detected in plasma tau and p-tau181levels between the two groups(P＞0.05).The plasmaα-syn level was positively related with the FSS score(r=0.237,P=0.030)and was an influencing factor of FSS(OR=1.019,95％CI:1.006-1.032,P=0.004).Conclusion Plasma neurodegenerative protein level is related with NMS and plasmaα-syn level is a peripheral biomarker for fatigue in PD patients.

PURPOSE: Research has shown that sevoflurane-induced toxicity causes neurodegeneration in the developing brain. miR-34a has been found to negatively regulate ketamine-induced hippocampal apoptosis and memory impairment. However, the role of miR-34a in sevoflurane-induced hippocampal neurodegeneration remains largely unclear. MATERIALS AND METHODS: C57/BL6 mice (7-day-old) inhaled 2.3% sevoflurane for 2 h/day over 3 consecutive days. miR-34a expression was reduced through intracerebroventricular injection with miR-34a interference lentivirus vector (LV-anti-miR-34a) into mouse hippocampus after anesthesia on the first day of exposure. Hippocampal apoptosis was detected by TUNEL assay and flow cytometry analysis. Spatial memory ability was evaluated by the Morris water maze test. The interaction between miR-34a and Wnt1 was confirmed by luciferase reporter assay, RNA immunoprecipitation, Western blot, and immunofluorescence staining. The effects of miR-34a on protein levels of B-cell lymphoma 2 (Bcl-2), bcl-2-like protein 4 (Bax), and Wnt/β-catenin pathway-related proteins were evaluated using Western blot analysis. RESULTS: Sevoflurane upregulated hippocampal miR-34a, and miR-34a inhibitor attenuated sevoflurane-induced hippocampal apoptosis and memory impairment. miR-34a negatively regulated Wnt1 expression by targeting miR-34a in hippocampal neurons. Moreover, forced expression of Wnt1 markedly undermined miR-34a-mediated enhancement of sevoflurane-induced apoptosis of hippocampal neurons, while Wnt1 silencing greatly restored anti-miR-34a-mediated repression of sevoflurane-induced apoptosis of hippocampal neurons. Increased expression of miR-34a inhibited the Wnt/β-catenin pathway in hippocampal neurons exposed to sevoflurane, while anti-miR-34a exerted the opposite effects. CONCLUSION: miR-34a inhibitor may effectively protect against sevoflurane-induced hippocampal apoptosis via activation of the Wnt/β-catenin pathway by targeting Wnt1.
Anesthesia ; Animals ; Apoptosis* ; Blotting, Western ; Brain ; Flow Cytometry ; Fluorescent Antibody Technique ; Hippocampus ; Immunoprecipitation ; In Situ Nick-End Labeling ; Lentivirus ; Luciferases ; Lymphoma, B-Cell ; Memory ; Mice ; Neurons ; Repression, Psychology ; RNA ; Spatial Memory ; Water

Objective To evaluate the effect of isoflurane on the expression of phosphorylated glycogen synthase kinase-3 beta (p-GSK-3β) and β-catenin in neural stem cells (NSCs) in the hippocampus of developing rats.Methods Twenty-four 7-day-old Sprague-Dawley rats,weighing 15-20 g,were divided into 2 groups (n =12 each) using a random number table:control group (group C) and 2％ isoflurane group (group Ⅰ).Group C inhaled 30％ oxygen for 4 h.Group Ⅰ inhaled 2％ isoflurane in 30％ oxygen for 4 h.Six rats were randomly selected from each group,and 5-bromodeoxyuridine (BrdU) 200 mg/kg was intraperitoneally injected immediately before anesthesia to assess the proliferation of NSCs in the hippocampal dentate gyrus.The rats were sacrificed at 6 h after the end of anesthesia,and hippocampi were isolated for determination of the number of BrdU positive cells in the dentate gyrus (by immunohistochemistry) and expression of p-GSK-3β and β-catenin in hippocampal tissues (by Western blot analysis).Results Compared with group C,the number of BrdU positive cells was significantly decreased,and the expression of p-GSK-3β and β-catenin was down-regulated in group Ⅰ (P＜0.05).Conclusion Isoflurane can inhibit the proliferation of NSCs in the hippocampal dentate gyrus of developing rats,and the mechanism may be related to down-regulation of the expression of p-GSK-3β and β-catenin.

Objective To explore the application value of susceptibility weighted imaging(SWI)in measuring brain iron deposition in the diagnosis and the assessment of Parkinson’s disease(PD).Methods Thirty cases with PD underwent head routine magnetic resonance imaging and SWI scanning.The unified PD rating scale(UPDRS)was used to assess the severity of PD.The phase values of substantia nigra (SN),red nucleus(RN),caudate nucleus (CA),globus pallidus(GP)and putamen (PUT)were measured manually on the phase map.The correlation between the phase values of the region of interest(ROI)and the UPDRS scores of PD was analyzed.Results There were no significant differences between the phase values of the less severe body side and those of the more severe body side (SN,P=0.120;RN, P=0.402;CA,P=0.196;GP,P=0.616;PUT,P =0.985).Significant negative correlations were found between the phase values of SN, CA,GP and the UPDRS Ⅲ scores,respectively (SN-UPDRS Ⅲ:r=-0.407,P =0.026;CA-UPDRS Ⅲ:r =-0.424,P =0.02;GP-UPDRS Ⅲ:r=-0.363,P =0.048).Significant negative correlation was found between the phase values of SN and the UPDRSⅤranks (r=-0.373 ,P =0.043 ),while the similar correlation was found between the phase values of CA and the UPDRSⅠ scores (r=-0.367,P =0.046)and the medium negative correlation was found between the phase values of GP and the gait disorder scores of UPDRS Ⅲ(r=-0.41 1,P =0.024).But no correlation was found between the phase values of other ROIs and the UPDRS scores. Conclusion SWI could quantitatively assess the brain iron deposition of the PD patient,which provided reference for clinical diagnosis and assessment of PD.

Objective To evaluate the effect of isoflurane anesthesia on Wnt3a expression in the hippocampus of developing rats.Methods Twenty-four 7-day-old male Sprague-Dawley rats,weighing 125-155 g,were randomly divided into 3 groups (n=8 each) using a random number table:control group (group C),4 h inhalation of 2％ isoflurane group (group I4),and 6 h inhalation of 2％ isoflurane group (group I6).The rats were sacrificed at 6 h after the end of treatment in each group,and the hippocampi were removed for determination of Wnt3a mRNA expression (by real-time reverse transcriptase polymerase chain reaction) and Wnt3a expression (by Western blot).Results Compared with group C,the expression of Wnt3a and Wnt3a mRNA in hippocampi was significantly up-regulated in I4 and I6 groups (P＜0.05).Compared with group I4,the expression of Wnt3a in hippocampi was significantly up-regulated (P＜0.05),and no significant change was found in the expression of Wnt3a mRNA in hippocampi in group I6 (P＞0.05).Conclusion The mechanism of isoflurane-induced neurotoxicity is probably related to upregulation of Wnt3a expression in the hippocampal tissues of developing rats.