Objective:To study the effect of endothelin receptor antagonist BQ-123 on the nerve function damage after subarachnoid hemorrhage (SAH)in the rats,and to explore the mechanisms.Methods:Total 120 male SD rats were divided into sham group,SAH group,low dose of BQ-123 group (50 μg· kg-1 )and high dose of BQ-123 group (75 μg·kg-1 ).The SAH rat models were established by injecting the autologous blood into cisterna magna twice.The morphological changes of hippocampus nerve cells of rat brain tissue were detected with HE staining, and the expressions of mTOR, Beclin-1 and LC3-Ⅱ in the hippocampus of rats were detected with immunohistochemistry and RT-PCR;the shuttle-box experiment was used to evaluate the abilities of learning and memory,and the holding power evaluation was used to evaluate the forelimb pulling force of the rats in various groups at each time point.Results:Compared with sham group,the morphological damages of neurons of the rats in SAH group were increased,the survival rate of neurons of the rats in SAH group was decreased (P <0.05),the expression levels of mTOR mRNA,Beclin-1 mRNA and LC3-Ⅱ mRNA in hippocampus tissue of the rats were increased (P < 0.05),and the abtilities of learning and memory and the values of holding power were decreased (P <0.05).Compared with SAH group,the morphological damages of neurons of the rats in BQ-123 groups were decreased,the survival rates of neurons of the rats in BQ-123 groups were increased (P < 0.05),the expression levels of mTOR mRNA of rats were decreased (P <0.05),the expression levels of Beclin-1 mRNA and LC3-ⅡmRNA in hippocampus tissue were increased (P <0.05),and the abilities of learning and memory and the values of holding power were increased (P < 0.05 ). The changes were more significant in high dose of BQ-123 group compared with low dose of BQ-123 group (P <0.05).Conclusion:BQ-123 can improve the nerve function damage after SAH in the rats,its mechanism may be related to regulating the mTOR/autophagy signaling pathway.

Objectives To investigate the treatment effect of endothelin A receptor antagonist BQ-123 on early brain injury of subarachnoid hemorrhage (SAH ) in rats and its mechanism. Methods According to the random number table method,120 SD rats were divided into four groups:a sham operation (sham),a SAH,a high-dose BQ-123 (75 μg/ kg),and a low-dose BQ-123 (50 μg/ kg)(n = 30 in each group). A rat model was induced by using the injection of blood into cisterna magna twice. After establishing models at hours 6,24,72,and 144,the rats were further divided into four subgroups. Light and electron microscopes were used to observe the changes of the morphological structure in hippocampal area. Immunohistochemistry and RT-PCR were used to detect the expression levels of phosphoinositide 3 kina (PI3-K),protein kinase B (PKB/ Akt),and mammal target of rapamycin (mTOR). Results (1)In the process of model making,7 rats died and 1 model did not meet the criteria and was excluded from the SAH group. Six rats died in the high-dose BQ-123 group and the low-dose BQ-123 group respectively. One rat in each group did not meet the criteria and was excluded. The rats were included in the final statistical analysis:30 in the sham group,22 in the SAH group,23 in the high-dose BQ-123 group,and 23 in the low-dose BQ-123 group. (2)Compared with the sham group,the expression levels of PI3-K,AKt and mTOR were increased signifi-cantly (all P < 0. 05). Compared with SAH group,the hippocampal neuronal morphology and structure damage were alleviated in the low-dose BQ-123 group. The number of surviving neurons at each time point was increased ([132 ±18],[110 ±16],[84 ±13],[92 ± 10]cells/ HP,all P < 0. 05). The tensile force values of rats were increased at each time point and the learning and memory function were improved. The expression levels of PI3-K and Akt were further increased (all P < 0. 05). The expression level of mTOR was decreased (all P < 0. 05). (3)Compared with the low-dose BQ-123 group,the morphological and structural damage of hippocampal neurons were alleviated. The number of surviving neurons at each time point ([153 ±20],[131 ± 18],[137 ±19]and [135 ± 17]cells/ HP)was increased (all P < 0. 05). The tensile force values of the rats were increased at each time point. The learning and memory function of the animals were improved. The expression levels of PI3-K (3. 8 ± 0. 8,8. 9 ± 2. 4,8. 6 ± 2. 4,and 6. 2 ± 2. 0)and Akt (4. 86 ± 1. 74, 8. 64 ± 1. 62,7. 94 ± 1. 70,and 6. 48 ± 1. 58)were further increased (all P < 0. 05). The expression levels of mTOR (2. 89 ± 0. 26,2. 14 ± 0. 18,1. 94 ± 0. 17,and 1. 62 ± 0. 12)were decreased (all P < 0. 05). Conclusions BQ-123 has as a good therapeutic effect for early brain injury after SAH. Its mechanism may be associated with the regulation of PI3-K/ Akt signaling pathway.

Objective To study the effects of 3-n-butylphthalide (NBP) on motor function and learning and memory ability in rats with diffuse brain injury (DBI). Methods 128 male Sprague-Dawley rats were randomly divided into sham operation group, traumatic group,low-dose NBP treatment group and high-dose NBP treatment group with 32 rats in each group. Every group was divided into 24 h, 48 h and 72 h subgroups. DBI rat model was established according to the description of Marmarou's diffused brain injury. The histopathologic changes of cerebral tissue (0.2 cm bilateralis coronal line) were observed by light and electron microscope at 24 h, 48 h and 72 h after injury. Morris water maze and rolling test were performed daily at 24, 48 and 72 h (time points). Results Compared with the model group, the damage of brain tissue decreased, and the survival nerve cells increased (P<0.01). The behavioral tests showed that the latency to find the platform shortened (P<0.01) and the frequency of crossing the platform increased in the 72 h subgroup (P<0.01). The general movement ability enhanced in NBP groups (P<0.05). All the indexes were more significant in high-dose NBP treatment group. Conclusion NBP can improve neurological function and learning and memory function after brain injury and the molecular mechanisms is related to the decrease of the nerve cells lose and traumatic cerebral ultrastructure damage.

Objective To investigate the relationship of extracellular regulated protein kinases activation and neural cells autophagy in rats after subarachnoid hemorrhage.Methods One hundred twenty male SD rats were randomly divided into sham operated group,SAH group,ERK1/2 inhibitor U0126 group,autophagy inducer rapamycin (Rap) group.The animal models were established by injecting the autologous blood into cisterna magna twice.U0126 (5μ g/μL) and Rap (10nmol/μL) were injected into lateral ventricles in U0126 group and Rap group 30min before SAH.The morphology of hippocampal nerve cells were examined by using light microscopy.The expression levels of phosphorylated ERK 1/2 (p-ERK 1/2),ERK 1/2mRNA and autophagy markers (Beclin-1 and Beclin-1 mRNA、LC3-Ⅱ and LC3mRNA) in the hippocampus were detected by using inmunohistochemistry and real-time fluorescence quantitative PCR.Result Compared with sham group,the rate of dead nerve cells,the mRNA levels of ERK1/2,Beclin-1 and LC3 as well as the levels of the p-ERK1/2,Beclin-1 and LC3-Ⅱ increased in SAH group (P＜0.05).Compared with SAH group,the rate of dead nerve cells increased(P＜0.05),the ERK1/2 mRNA,Beclin-1 mRNA and LC3 mRN A,and p-ERK1/2,Beclin-landLC3-Ⅱ in U0126 group decreased(P＜0.05);the rate of dead nerve cells decreased (P＜0.05),the Beclin-1 mRNA and LC3 mRNA,the Beclin-1and LC3-Ⅱ level increased in Rap group(P＜0.05),but ERK 1/2 mRNA and p-ERK 1/2 remained unchanged (P＞0.05).Conclusion Activation of the ERK1/2 signaling pathway after SAH,can induce nerve cells death by increasing Beclin-1 and LC3-Ⅱ expressions.

Objective To investigate the effect of MAPK activation on autophagy in the hippocampus of rats with subarachnoid hemorrhage.Methods A total of 100 male SD rats were divided into 5 groups randomly:sham operated group,SAH group,inhibitor U0126 group,inhibitor SB203580 group,SP600125 group.The animal model was established by injecting the autologous blood into cisterna magna twice.The morphological changes of hippocampus nerve cells of rat brain were detected with HE.The mRNA levels of ERK1/2,p38MAPK,JNK and LC3 in hippocampus were detected with quantitative real time PCR and the expression of phosphorylated ERK1/2,phosphorylated p38MAPK,phosphorylated JNK and LC3-Ⅱ in hippocampus of rat brain were detected by immunohistochemistry.Results Compared with the Sham group,the survival rate of neurons in SAH group decreased (6 h:(84.982 ± 5.723) ％,24 h:(74.383± 9.860) ％,48 h:(62.860± 10.820) ％,72 h:(52.260± 10.960) ％) (all P＜0.05).The levels of ERK1/2 mRNA,p38MAPK mRNA,JNK mRNA and LC3 mRNA in hippocampus increased (all P＜ 0.05) and the expression of p-ERK1/2,p-p38MAPK,p-JNK,LC3-Ⅱ proteins increased(all P＜0.05).Compared with the SAH group,the survival rate of neurons in U0126 group was decreased (6 h:(71.620±6.542) ％,24 h:(66.221±7.742)％,48 h:(55.208±8.802) ％,72 h:(46.242±7.782) ％),and the ERK1/2 and LC3 in hippocampus decreased both in mRNA level and in protein level(all P＜0.05).Compared with the SAH group,the survival rate of neurons in SB203580 groups was increased (6 h:(89.082±6.602)％,24 h:(85.840±9.726) ％,48 h:(74.96± 10.916) ％,72 h:(69.211 ± 10.745) ％),and the p38MAPK,LC3-Ⅱ in hippocampus decreased at both mRNA and protein levels (all P＜0.05).Compared with the SAH group,the survival rate of neurons in SP600125 groups was increased (6 h:(91.620± 7.542) ％,24 h:(86.221 ± 10.742) ％,48 h:(75.208±11.802) ％,72 h:(70.242± 11.782) ％).The expression of JNK was decreased while the LC3-Ⅱ was increased in hippocampus (P＜0.05).Conclusion MAPK activation is involved in the autophagy of hippocampal neurons after SAH,in which ERK1/2 activation plays a positive role in the regulation of autophagy in hippocampal neurons after SAH,while p38MAPK and JNK activation plays a negative role in autophagy.

Objective To explore the expression levels of immune-related microRNA-146b (miR-146b),microRNA-155 (miR-155) and microRNA-30b(miR-30b) in human breast milk and its relationship with maternal and infant's health.Methods One hundred and thirty-four mothers and their infants from obstetrical department were recruited in the study after delivery.The subjects were divided into 2 groups,breast feeding group(n =86) and formula-feeding group(n =48),and were followed up 3 months after delivery.Breast milk samples were collected at 2-5 days after delivery(colostrum) and 3 months after delivery(mature milk).The expression levels of microRNAs in milk samples were detected by real-time PCR.The relationship between levels of microRNAs and maternal and infant-related factors was analyzed.Results 1.MiR-146b,miR-155 and miR-30b expressions were abundant both in human colostrums (5.950 ± 0.823,3.899 ± 0.920,4.057 ± 0.604) and mature milk (4.840 ± 0.805,2.128 ± 0.969,4.929 ± 0.566).The levels of miR-146b and miR-155 were higher in colostrum than that of mature milk (t =7.716,10.215,all P ＜ 0.01),while the level of miR-30b was higher in mature milk than that of colostrums(t =-8.626,P ＜ 0.0l).2.Additionally,the level of miR-30b was negatively correlated with maternal pre-pregnancy body mass index (r =-0.298,P ＜ 0.01).3.The levels of miR-146b and miR-30b were higher in mothers giving birth by vaginal delivery than those who underwent cesarean section(t =2.356,3.108,all P ＜0.05).4.The levels of miR-146b and miR-155 were higher in colostrum-fed girls than boys (t =-2.204,-2.985,all P ＜ 0.05).5.The level of miR-146b in mature milk was negatively correlated with 3-month-old infant' s Z score of body weight (r =-0.425,P ＜ 0.05) and body length (r =-0.569,P ＜ 0.01).6.During follow-up,the incidence of baby eczema in breast feeding group (8.82％,3/34 cases) was lower than that in formula milk feeding group(29.17％,14/48 cases) (x2 =5.012,P =0.025).Conclusions The levels of immunocompetent microRNAs in human milk are influenced by the lactation period,maternal prepregnancy body mass index,mode of delivery and infant sex.The immune-related microRNAs in human milk could be involved in the regulation of infant's immunity and growth.

Objective To analyze the distribution and antibiotic resistance of bacterial pathogens in blood culture in the Department of Hematology of the First Affiliated Hospital of Nanjing Medical Universi-ty from 2013 to 2017. Methods BACTEC FX automatic blood culture instrument was used for blood cul-ture. VITEK-2 Compact automatic microbial identification and drug susceptibility system was used for bacte-rial identification, drug sensitivity testing and fungal identification. K-B and ATB FUNGUS 3 methods were used for Streptococcus and fungi susceptibility testing, respectively. WHONET5. 6 software was used for sta-tistical analysis. Results The positive results of blood culture accounted for 7. 6% during 2013 to 2017 and the top five isolated pathogenic bacteria were Escherichia coli (20. 9% ), Klebsiella pneumoniae (18. 0% ), coagulase negative Staphylococcus (12. 4% ), Pseudomonas aeruginosa (11. 5% ) and Enterobacter cloacae (3. 6% ). Carbapenems (94. 4% ) were the most active antibiotics against Enterobacteriaceae. Pseudo-monas aeruginosa strains were sensitive to all antibacterial dugs except aztreonam. Acinetobacter baumannii strains had a lower sensitivity to almost all antibacterial dugs. Vancomycin, linezolid and teicoplanin were active antimicrobial agents against gram-positive cocci. Conclusion Patients with hematological diseases are more susceptible to nosocomial infection, which should be paid more attention to in clinical and laborato-ry practice. Rational antimicrobial therapy is an effective way for control of antimicrobial resistance.

Objective To investigate synergistic effects of extracellular signal regulated kinase (ERK1/2) and phosphatidylinositol 3 kinase (PI3-K) pathway inhibitors on autophagy in the hippocampus of rats with subarachnoid hemorrhage (SAH),for identification of therapeutic targets in SAH.Methods Totally,200 male SD rats were randomly divided into a sham operated group,SAH group,inhibitor U0126 group,inhibitor LY294002 group,and a U0126+ LY294002 group.Animal models were established by injecting autologous blood twice into the cisterna magna.Morphological changes in the hippocampus nerve cells were detected by HE staining;ERK1/2,PI3-K,beclin-1,and LC3 mRNA expression in the hippocampus were detected by real-time PCR,and phosphorylated ERK 1/2,PI3-K,beclin-1,and LC3-Ⅱ protein expression were detected by immunohistochemistry.Results Neuronal death rate and phosphorylated ERK1/2,PI3-K,beclin-1,and LC3-Ⅱ levels in the hippocampus in the SAH group were higher than in the sham group (all P ＜ 0.05).Neuronal death rate in U0126 or LY294002 group was higher than in SAH group,while ERK1/2,PI3-K,beclin-1,and LC3 mRNA and phosphorylated ERK 1/2,PI3-K,beclin-1,and LC3-Ⅱ protein levels were lower than in SAH group (all P ＜ 0.05).Neuronal death rate in U0126 +LY294002 group was higher than in U0126 or LY294002 group,while ERK1/2,PI3-K,beclin-1,LC3 mRNA and phosphorylated ERK 1/2,PI3-K,beclin-1,and LC3-Ⅱ protein levels in the hippocampus were lower than in U0126 or LY294002 group (all P ＜ 0.05).Conclusion Co-targeting the inhibition of ERK 1/2 and PI3-K pathways can significantly reduce neuronal cell autophagy and aggravate cells loss after SAH.

Objective:To investigate the myocardial protective effect of extracorporeal cardiac shock wave therapy (CSWT) combined with sulfur hexafluoride microbubble post-conditioning on myocardial ischemia-reperfusion injury (MI/RI) in rats, and to provide theoretical support for clinical treatment of MI/RI.Methods:A total of 32 male SD rats were randomly divided into 4 groups: sham operation group (Sham group), MI/RI group (IR group), CSWT group (IR+ SW group), and CSWT combined with sulfur hexafluoride microbubble treatment group (IR+ SW+ MB group), with 8 rats in each group. Therapeutic intervention was performed in IR+ SW group and IR+ SW+ MB group on the 1st, 3rd and 5th day after modeling. The left ventricular end-diastolic diameter (LVEDD), left ventricular end-systolic diameter (LVESD), left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) of the rats were measured by echocardiography before and after treatment. On the 7th day, myocardial fibrosis was assessed by Masson staining, and cardiomyocyte apoptosis was observed by TUNEL staining. The myocardial apoptotic proteins Bcl-2, BAX, Cleaved-Caspase-3 and Cleaved-Caspase-9 in the infarct boundary area were detected by Western blot. The differences of the above indexes among different groups were compared.Results:①There was no significant change in heart rhythm and heart rate among the groups before and after treatment, and there was no significant difference in heart rate ( P>0.05). ②The echocardiographic results after treatment showed that, compared with IR group, LVEDD and LVESD in IR+ SW group and IR+ SW+ MB group decreased in turn, while LVEF and LVFS increased in turn with significant differences between each two groups (all P<0.05). ③Compared with IR group, the degrees of myocardial fibrosis and apoptosis in IR+ SW group and IR+ SW+ MB group were alleviated in turn, and the relief in IR+ SW+ MB group was the most obvious. Quantitative analysis showed that compared with IR group, the proportions of cardiomyocyte apoptosis in IR+ SW group and IR+ SW+ MB group decreased in turn, and there were significant differences between each two groups (all P<0.05). ④The results of Western blot detection showed that compared with IR group, the levels of Bcl-2 in IR+ SW group and IR+ SW+ MB group increased in turn, while the levels of BAX and the activation level of Caspase-3 and Caspase-9 protein decreased in turn. These differences were all statistically significant between each two groups (all P<0.05) except for the activation level of Caspase-3 protein between IR+ SW group and IR+ SW+ MB group ( P>0.05). Conclusions:CSWT combined with sulfur hexafluoride microbubble therapy can improve left ventricular remodeling and left ventricular systolic function by inhibiting cardiomyocyte apoptosis.

Objective:To explore the effect of Plantaginis semen on the expressions of AQP4 gene and protein in small intestine of diarrhea rats. Methods:Sixty Sprague-Dawley male rats were randomly divided into normal control group, model control group, hydrochlorothiazide group, and low-, medium-, and high-dose Plantaginis semen groups. Intragastric administration of Senna (20 ml/kg) was implemented in the morning for 5 groups except the normal group. The rats in the low-, medium-, and high-dose Plantaginis semen groups were intragastrically administered with 0.95, 1.9, 3.8 g/kg Plantaginis semen formula granule solution, while the rats in hydrochlorothiazide group were given hydrochlorothiazide suspension 9 mg/kg by gavage according to 1 ml/100 g body weight for 14 days. The loose stool rate, average ranking of loose stool, and diarrhea index were compared according to the fecal traits and stool times after 14 days of treatment, and the small intesine tissue were collected. Hematoxylin-eosin (HE) staining to observe the pathological morphological changes in small intestine, and Quantiative Real-time PCR and Western blot were applied to detect the gene and protein expressions of AQP4. Results:Compared with the model group, the loose stool rate (50.89% ± 6.17%, 41.14% ± 4.48%, 36.37% ± 4.81 % vs. 67.45% ± 7.35%), the average ranking of loose stool (2.16 ± 0.34, 1.73 ± 0.28, 1.52 ± 0.25 vs. 2.63 ± 0.29), and the diarrhea index (1.10 ± 0.19, 0.71 ± 0.11, 0.57 ± 0.12 vs. 1.77 ± 0.24) of rats in each group of Plantaginis semen significantly decreased ( P<0.05); the degree of intestinal mucosal injury, hyperemia and neutrophil infiltration were alleviated; the expressions of AQP4 mRNA (0.48 ± 0.10, 0.69 ± 0.12, 0.97 ± 0.15 vs. 0.21 ± 0.03), and the protein of AQP4 (0.59 ± 0.08, 0.64 ± 0.09, 0.78 ± 0.11 vs. 0.32 ± 0.05) in the small intestine tissue of Plantaginis semen groups significantly increased ( P<0.05). Conclusions:Plantaginis semen has antidiarrheal effect, and its mechanism is related to up-regulation the gene and protein expressions of AQP4, addition of water absorption and promotion of water and fluid metabolism.