AIM:To explore the mechanism by which Chaijin-Jieyu-Anshen tablet(CJJY)-medicated serum prevents synaptic injury in rat anterior cingulate cortex(ACC)neurons using an in vitro depression model.METHODS:Cells(astrocytes,microglia and neurons)were isolated from the ACC of SD rats.The isolated cells were characterized by immunofluorescence staining.An in vitro depression model was developed using 1 mg/L lipopolysaccharide(LPS)com-bined with 200 μmol/L corticosterone(CORT).These cells were divided into control group,model group(CORT+LPS),glucocorticoid receptor(GR)blocker(GR-)group(CORT+LPS+RU486),GR agonist(GR+)group(CORT+LPS+dexa-methasone),CX3C chemokine receptor 1(CX3CR1)blocker(CX3-)group(CORT+LPS+AZD8797),CX3CR1 agonist(CX3+)group(CORT+LPS+fractalkine),CJJY group(CORT+LPS+CJJY-medicated serum),CJJY/GR+group(CORT+LPS+CJJY-medicated serum+dexamethasone),and CJJY/CX3+group(CORT+LPS+CJJY-mediated serum+fractalkine).The morphological characteristics of all ACC cells were observed by high-content analysis.The levels of neuroendocrine-related factors,adrenocorticotropic hormone(ACTH),corticotropin-releasing hormone(CRH)and CORT,and neuroin-flammatory mediators,tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),IL-6 and glutamate(Glu),in the cell supernatants were quantified by ELISA.Immunofluorescence staining was used to analyze the protein expression of GR and vesicular glutamate transporter 1(VGluT1)in astrocytes,as well as CX3CR1 and adenosine A2A receptor(A2AR)in microglia.Immunofluorescence staining with Nissl and β-tubulin was performed to evaluate synaptic damage in ACC neurons.RESULTS:In an in vitro model of depression,CJJY-medicated serum prevented morphological damage to ACC neurons,microglia and astrocytes.Moreover,CJJY-medicated serum reversed abnormal increases in the levels of ACTH,CRH,CORT,TNF-α,IL-1β,IL-6 and Glu in cell supernatants(P<0.05 or P<0.01).It was also found that CJJY-medi-cated serum reduced abnormal expression of GR,VGluT1,CX3CR1 and A2AR(P<0.05 or P<0.01),alleviating damage to the neuronal dendrites and dendritic spines of ACC neurons.CONCLUSION:The CJJY-medicated serum regulates GR/CX3CR1 double signaling in glia,and attenuates rat ACC neuronal synaptic damage in an in vitro depression model,indicating that CJJY-medicated serum controls depression by affecting the GR/CX3CR1 double signaling.

Objective: To investigate the changes of brain activity in adolescents with first-episode major depressive disorder (MDD) and its significance. Methods: Totally 42 first-episode MDD patients with suicidal ideation (SU group), 17 MDD patients without suicidal ideation (NSU group) and 29 healthy controls (HC group) were examined by psychological assessment and resting-state functional magnetic resonance imaging (fMRI). The differences of brain functional activities between the three groups were compared by amplitude of low frequency fluctuation(ALFF). The relationship between the difference of zALFF value in brain regions and the severity of depression, intensity of suicidal ideation were further analyzed. Results: The zALFF values in the left hippocampus(MNI: x=-21, y=-33, z=0) and left Frontal_Sup_Medial(MNI: x=3, y=33, z=63) of the three groups were significantly abnormal (P<0.001, k>20, uncorrected). The results of Post-hoc test showed that the zALFF value of left hippocampus increased significantly (F=28.931) and decreased significantly in left Frontal_Sup_Medial(F=16.648)in SU group , compared with NSU group and HC group(P<0.001, Bonferroni correction). There was no significant difference in zALFF value between HC group and NSU group(P>0.05). In SU group, the zALFF values of left hippocampus and left Frontal_Sup_Medial were not correlated with total scores of HAMD-24(r=-0.214, -0.103) and suicide ideation intensity factor scores(r=0.115, -0.055)(P>0.05). Conclusion Abnormal autonomic nervous activity exists in left hippocampus and left Frontal_Sup_Medial in adolescent MDD patients with suicidal ideation, which may be the neurological basis and biological index of suicidal ideation.

Objective To compare the biological characteristics of several different anxiety rat models established by different methods of stress at different time points and provide experimental basis for the most appropriate modeling methods .Methods 60 rats were randomly divided into normal , empty bottle stress , chronic emotional stress ( CES ) group, restraint stress for 3h, 6h, and modeling respectively .In the experimental 7 d, 14 d, 21 d, elevated plus maze and fear condition system was used to test anxiety-like behavior in rats , open field test to study anxiety or depression-like behavior , forced swimming test was used to detect depression-like behavior in rats , and using the Elisa test kit to detect the contents of 5-HT, DA in the hippocampus in rats .Results Anxiety-like behavioral test results showed that rats in empty bottle stress, CES, 6 h restraint stress group started to have anxiety-like behavior since 14 d, then anxiety-like behavior was becoming increasingly apparent .Forced swimming test results showed that immobility time in 6 h restraint rats was significantly increased in the first 7 d(P <0.05).Meanwhile, compared with control group, hippocampal 5-HT, DA contents in empty bottle stress and CES rats increased significantly since 14 d.Conclusions Among several stress methods established anxiety model , anxiety-like behavior in 3 h restraint stress was not obvious; 6 h restraint stress exhibited a depression-like behavior in the forced swimming test might be due to prolonged stress .Empty bottle stress and CES can successfully establish the anxiety rat model , and the anxiety behavior of the rats have some differences . Corresponding model methods can be selected according to different experimental purposes .

Objective To study the content of monoamine neurotransmitters and neurotrophic factor in the hippocampus, amygdala and prefrontal cortex in anxious depression rats, and explore the possible pathogenesis.Methods 60 SD rats were randomly divided into normal group, vehicle group, anxiety group, depression group, and anxious depression group, 12 rats in each group.Chronic restraint stress combined with corticosterone injection was used to establish anxiety and depression model, the modeling time was 21 d.After modeling, elevated plus maze test, open field test, and forced swimming test were used to evaluate the anxiety and depression-like behavior, HPLC-ECD was used to detect the content of 5-HT, NE, and DA in the hippocampus, amygdala, and prefrontal cortex of rats.Western-blotting was used to detect the expression of BDNF and NT-3 in rats.Results Rats in anxious depression model group were comparable to the anxiety group in time and frequency entering open arm time, and number of locomotor activity in open field, and it had a significant difference when compared with the control and depression groups (P<0.01 or P<0.05).Immobile time in anxious depression model rats was increased significantly when compared with the control and anxiety groups (P<0.01).Meanwhile, compared with the control group, 5-HT in hippocampus and 5-HT, NE in amygdala or prefrontal cortex were significantly decreased in the depressive rats with anxiety (P<0.01 or P<0.05).Moreover, the content of BDNF and NT-3 was significantly decreased in each brain regions compared with the control group (P<0.01 or P<0.05), and BDNF levels were obviously decreased compared with the anxiety group (P<0.05).Conclusions Rats of anxious depression have significant anxiety and depression-like behaviors.Its mechanism may be associated with the down-regulation of monoamine neurotransmitters and neurotrophic factors BDNF and NT-3 in hippocampus, amygdala, and prefrontal cortex region.

Objective To investigate the effects of dizocilpine(MK801) on depressive-like behaviors and damaged hippocampus in rats with diabetes-related depression.MethodsThe animal models of diabetes-related depression were established and they were randomly divided into two groups based on random number table: model group and MK801 group,while 6 rats were included in each group.And another six health rats were regarded as control group.The Open-field test was used to detect the activities.The damage of hippocampus was valued by HE staining,Nissl staining,and Tunel staining.The protein expressions of Bax,Bcl-2 in hippocampus were detected by Western blot.ResultsThe number of activities was significant decreased in Open-field test in model group when compared with control.Hippocampal neurons vacuoles,Nissl bodies were decreased and apoptotic cells were increased in hippocampus in model group as well.Furthermore,the expression of Bax was significant up-regulated(94.57±7.97,P<0.01),while the Bcl-2 was declined(24.65±5.26,P<0.01).Compared with the model group,the animals in MK801 group exhibited increased activities(12.50±4.42,P<0.01),which accompany with an increased Nissl body(133.55±16.74,P<0.01) and a decreased apoptosis(22.50±6.35,P<0.01).Moreover,the expression of Bax was decreased and the Bcl-2 was increased in MK801 group when they were contrasted to model(33.00±4.57,P<0.01).Conclusion MK801 is a significant element to regulate the expression of apoptosis protein including Bax,Bcl-2,and to protecte the hippocampal neuron in rats with diabetes-related depression effectively.

Objective To investigate the effects of Zuogui Jiangtang Jieyu Prescription on neurotrophic effects of astrocytes in diabetes mellitus rats with depression. Methods The diabetes mellitus with depression rat models were established by composite method, and then were randomly divided into 5 groups: model group, positive group, Zuogui Jiangtang Jieyu Prescription high-, medium-, low-dose groups, with 12 rats in each group. 12 normal rats were set as control group. Each administration group was given relevenat medicine for gavage for continuous 4 weeks. Open-field test was used to evaluate the depression-like behavior. The expression of GFAP in astrocyte and MAP2 in neuron were tested by immunohistochemistry. The expression of protein and mRNA of BDNF, GDNF, and NGF were tested by Western blot and RT-PCR. Results Compared with the control group, the activities of rats in model group were significantly reduced. The expression of GFAP increased, while the expressions of MAP2, BDNF, GDNF and NGF decreased. Compared with the model group, the depression-like behavior of rats in model group were significantly improved. The expression of GFAP decreased, while the expressions of MAP2, BDNF, GDNF and NGF increased, and GFAP decrseaed significantly. Conclusion The secretion function of astrocyte can be improved by Zuogui Jiangtang Jieyu Prescription. Its anti-depression and neuron-protection function might be correlated with the enhancement of neurotrophic effects of astrocytes.

Objective To observe the effects of Zuogui Jiangtang Jieyu Formula (ZJJF) on the ability of learning and memory and the expressions of JNK, Bcl-2 and Bax in hippocampus in diabetic rats with depression; To explore the protective mechanism of hippocampal damage in diabetic rats with depression. Methods High-fat gavage combined with intravenous injection of STZ was used to establish the model of diabetic rats. 28 days of chronic stress was given continuously and diabetic rats complicated with depression were built successfully. Then rats were randomly divided into 6 groups, including normal, model, positive medicine, high-, medium-, and low-dose of ZJJF groups. After the last administration, Morris water maze was used to detect escape latency time;Western blot was used to disclose the protein expressions of JNK, Bcl-2 and Bax in rat hippocampaus;RT-PCR was used to test the gene expressions of JNK and Bcl-2 and Bax. Results Compared with the normal group, escape latency time in model rats was significant longer (P<0.01), the protein and gene expression of JNK and Bax in rat hippocampaus significantly increased, Bcl-2 was markedly decreased (P<0.05, P<0.01);Compared with the model group, escape latency time in positive medicine group and high-dose of ZJJF group was significant shorter (P<0.01), the protein and gene expressions of JNK and Bax significantly decreased, and Bcl-2 markedly increased (P<0.05, P<0.01). Conclusion ZJJF can significantly improve the ability of learning and memory in diabetic rats with depression, which might be associated with preventing neuronal apoptosis in hippocampus.

Objective To explore the chiaical value of cerebral perfusion reduction after subarachnoid hemorrhage(SAH)by multi spiral CT perfusion in rabbits.Methods 32 male Japanese big-ear rabbits of clean level were randomly divided into 2 groups:Group A(n =8),the normal group,were to obtain normal MS-CTP values of rabbit cerebrums;Group B(n =24),the SAH group was made into two-hemorrhage animal models,checked on seven day by multi spiral CT perfusion.The source data was transmitted to the post-processing workstation ADW 4.2.Perfusion parameters maps(CBF)of both sides of frontal,parietal and basal ganglia were got by computing.Each rabbit was killed immediately after scanning,and the spider blood was observed.Results The average CBF value of the two groups:The difference was not statistically significant at the frontal(t =1.740,P =0.092)and parietal(t =1.868,P =0.072); The average values of CBF were significantly decreased at the basal ganglia(t =2.481,P =0.019).Group B showed the distribution of blood clot in the basal cistern,but there was no blood clot at the frontal and parietal.Conclusion The monitor perfusion changes of rabbit brain after SAH could be detected by MS-CTP,and the lower of regional cerebral perfusion had a relationship with the distribution with the blood clot,and it could instruct the early diagnosis of cerebral perfusion,and the clinical benefit to early intervention treatment.

Objective To investigate the value of permeability surface (PS) in predicting hemorrhagic transformation (HT) in acute ischernic stroke (AIS) using CT peffusion (CTP).Methods The study included 31 consecutive patients who presented symptoms suggestive of an AIS for 3-9 h. All patients underwent CT examination (noncontrast CT,CTP).HT was determined by follow-up CT images.According to presence of HT,the AIS was divided into HT group (PSHT,11 patients) and non-HT group(PSNo-HT,20 patients).PS,cerebral blood flow (CBF),cerebral blood volume (CBV) and mean transit time (MTT) on both sides of brains were measured.The relative PS(rPS),relative CBF (rCBF),relative CBV(rCBV) and relative MTT(rMTT) were obtained by calculating the ratio of the values of bilateral regions.The rPS between PSHT and PSNo-HT was compared with an exact Wilcoxon signed-rank test. The rCBF,rCBV,rMTT and the PS of the ischemic side between PSHr and PSNo-HT were compared with independent-sample t test.Meanwhile,Spearman rank correlation analysis was conducted to analyze the relationship between the CTP parameters and HT.ResultsThe PS value of ischemic side was (1.61±0.77) ml · min - 1 · 100 g-1 for the PSHT group,and the value was (0.91 ± 0.49) ml · min - 1 · 100 g- 1 for the PSNo-HT group.For the PSHT group,rPS,rCBF,rCBV,rMTT were 2.76 ±0.78,0.32 ±0.18,0.66 ±0.31,2.67 ±0.71,and for the PSNo-HT group,rPS,rCBF,rCBV,rMTT were 1.35 ±0.19,0.50±0.21,0.91 ±0.28,2.62 ± 1.31.Compared with PSNo-HT,PSHT had higher rPS and PS value,and there were significant statistical differences (U =0.000,t =3.070,P ＜0.01).But rCBF and rCBV values were lower in the PSHT group compared to the PSNo-HT group,and there were significant statistical differences (trCsF =2.343,trCBV =2.210,P ＜ 0.05).There was no significant statistical difference in rMTT between the two groups(t =0.118,P ＞ 0.05).Significant positive correlations were detected between the rPS and PS with HT(r=0.496,0.821,P ＜0.01).ConclusionsThe value of rPS is helpful in predicting HT in AIS.And it can be used as a predictor in determining clinical personalized treatment and thus reduce the incidence of adverse events.

Objective To evaluate perfusion computed tomography in the assessment of portal vein pressure changes in an experimental dog model of liver cirrhosis and portal hypertension.Methods The canine model of cirrhosis and portal hypertension was induced by portal vein stenosis with combination of systemic thioacetamide(TAA) feeding in drinking water.All of the Beagles in control group and cirrhotic group underwent hepatic perfusion on a spiral CT scanner.The parameters of hepatic perfusion were calculated by the method of deconvolution.The portal vein pressure was measured by a laparotomy surgery.Results ① In control group, the portal vein pressure was ( 14.5 ± 2.2) cm H2O, while it was (23.1 ± 2.8) cm H2O in PHT group, there was significant difference in the portal vein pressure between the two groups (P＜0.05).② The blood flow(BF) was (112 ±14) ml·100 g-1·min-1 in controls, while ( 96 ± 11) ml·100 g-1·min-1 in PHT group; the blood volume ( BV ) in control group and PHT group was (10 ±3) ml·100 g-1 and (11 ± 5) ml· 100 g-1, respectively; the mean transit time( MTT) was (7.1 ± 2.0) s and (10.4 ± 3.5) s, respectively; the hepatic arterial fraction (HAF) was ( 24 ± 5) % and ( 37 ± 6)% , respectively; the hepatic arterial perfusion (HAP) was(27 ±6) ml·100 g-1·min-1 and (35 ±5) ml·100 g-1·min-1, respectively; the portal venous perfusion (PVP) was (85 ± 13) ml·100 g-1·min-1 and (61 ±11) ml·100 g-1·min-1, respectively.There was significant difference in all parameters between the two groups except the parameter BV(P ＜ 0.05).③ In PHT group, the PVP and BF were negatively correlated with the portal vein pressure, while positively correlated with MTT and HAF.Portal vein pressure was negatively correlated with PVP, the equation, Y = 36.624 -0.219X, was deduced with linear regression analysis, by which the portal vein pressure in PHT Beagles was ( 23.2 ± 2.4) cm H2O, which was correlated with the observed by laparotomy value (23.1 ± 2.8) cm H2O (r = 0.843, P ＜ 0.05).Conclusion CT perfusion is a new non-invasive and effective method for assessment of portal vein pressure.