Objective: To investigate the effect of cold self-blood cardioplegia with ulinastatin on immature myocardial cell apoptosis and protein expressions of Bcl-2, Bax in ventricular septal defect (VSD) infants.
Methods: A total of 60 infants received VSD repairing operation with cardiopulmonary bypass (CPB) in our hospital were summarized. The patients were randomly divided into 2 groups:Test group, the infants received cold self-blood cardioplegia with ulinastatin when aortic cross-clamp was closed. Control group, the infants received cold self-blood cardioplegia when aortic cross-clamp was closed. n=30 in each group. The right atrium tissue was collected before CPB and 10 min after releasing aortic cross-clamp. The index of myocardial cell apoptosis was observed by TUNEL method, and the protein expressions of Bcl-2, Bax were examined by immunohistological method.
Results: Both groups showed the higher index of myocardial cell apoptosis at 10 min after releasing aortic cross-clamp than 5 min before CPB, and the apoptosis index in Test group was lower than that in Control group, all P<0.05. The protein expressions of Bcl-2 and Bax were obviously increased at 10 min after releasing aortic cross-clamp than 5 min before CPB in both groups. Compared with Control group, Test group presented the higher Bcl-2 protein expression and lower Bax protein expression, all P<0.05.
Conclusion: Cold self-blood cardioplegia with ulinastatin could protect immature myocardum from ischemia-reperfusion injury in VSD infants during CPB operation in clinical practice.

Objective To study the effect of single chain variable fragment (ScFv) antibody to EC3-4 fragment of desmoglein (Dsg) 3 in a mouse model of pemphigus vulgaris.Methods The ScFv an- tibody to EC3-4 fragment of Dsg-3 was injected subcutaneously into neonatal BALB/c mice at different time points;the mice were then evaluated clinically,histopathologically and by direct immunoflorescence exami- nation for the development of lesions.Results When injected alone,the ScFv antibody did not induce the appearance of key clinical features of pemphigus vulgaris.The antibody also did not prevent the develop- ment of pemphigus vulgaris features induced by sera of patients with pemphigus vulgaris,regardless of the time point of injection of ScFv antibody.Conclusion The ScFv antibody to EC3-4 fragment of Dsg-3 lacks pathogenicity in neonatal BALB/c mice,and also could not inhibit the development of lesions induced by sera from patients with pemphigus vulgaris.

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 observe the effects of Baishile Capsules on the hippocampal PI3K signaling pathway in chronic mild unpredictable stress (CUMS) rats; To discuss its mechanism of action for anti-depression. Methods SD rats were randomly divided into 6 groups, control group, model group, fluoxetine group, Baishile Capsules high-, medium- and low-dose groups, with 10 rats in each group. The cums depression model was established, and gavage for medication was conducted at the same time for 21 days in a row. The behavioral changes of rats in each group were detected by the novel feeding experiment and Open-field experiment; monoamine neurotransmitter in serum were detected by ELISA; the expressions of PI3K, AKT and SGK1 in rat hippocampus were detected by immunofluorescence and Western blot test. Results Compared with the control group, feeding latency of rats in model group was significantly prolonged (P<0.01), horizontal and vertical activities were significantly reduced (P<0.01); the content of serum monoamine neurotransmitter decreased (P<0.01); the expressions of PI3K and AKT in hippocampus decreased, while the expression of SGK1 increased (P<0.01, P<0.05). Compared with the model group, Baishile Capsule high- and medium-dose groups can significantly shorten the incubation period of feeding rats (P<0.05), and increase the number of horizontal and vertical activities (P<0.01); the 5-HT and NE levels in serum were significantly elevated (P<0.05, P<0.01); the expressions of PI3K, AKT in hippocampal increased; the expression of SGK1 decreased (P<0.05, P<0.01). Conclusion Baishile Capsules can alleviate the depression like behavior in rat models, and regulate key factors of hippocampal PI3K signaling pathway, so as to exert antidepressant effects.

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 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 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 investigate the effects of ulinastatin-containing autologous cold blood cardioplegic solution on the cardiac function of infants after cardiopulmonary bypass surgery.Methods Sixty infants younger than 10 months old,who underwent ventricular septal defect repair under cardiopulmonary bypass,were randomized into autologous cold blood cardioplegia group (30 patients,Group A)and ulinastatincontaining cold blood cardioplegia group (30 patients,Group B).CI,SI and LCWI were monitored 1 and 6 hours after opening the aorta.The time and rate of cardiac resuscitation,as well as the dependence on the inotropic drugs,were intraoperatively monitored.Results The automatic resuscitation rate in two groups was not siynificantly ( P ＞ 0.05).The time for automatic resuscitation were (34.2 ± 4.7) s and (52.1 ± 6.5 ) s for Group B and Group A,respectively ( P ＜ 0.05 ).The rate of dependence on inotropic drug were 40.0％ (12/30) and 66.7％ (20/30)for Group B and Gro~p A,respectively (P ＜ 0.05).Mter the operation,the CI,SI and LCWI of group B were higher than that of group A ( P ＜0.05 ).Conclusion Ulinastatin-containing autologous cold blood cardioplegic solution is beneficial to the functional cardiac recovery of the infants after heart bypass surgery by protecting the immature myocardium.

Objective To study the protective effect of intraaortic protamine injection on lung in infants undergwent opening heart operation by cardiopulmonary bypass surgery. Methods Sixty infants (age ≤ 1 year,weight ≤ 10 kg)who accepted opening heart operation by cardiopulmonary bypass surgery were randomly assigned into 2 groups ( n = 30 in each group) reciving intra-aortic and intra-venous protamine injection respectively. P-peak, P-plate, CL, Oxygenation Index, the number of WBC and neutrophil segregated in lungs were compared between two groups before injecting protamine and 10 minutes, 1 hour, 3 hours after injecting protamine. The time of mechanical ventilation were compared as well. Results P-peak, P-plate, the number of WBC and neutrophil segregated in lungs of intra-aortic injection group significantly decreased than intra-venous injection group at 1 hour, 3 hours after injecting protamine (t =2.743, 3.512; 3.218, 3.469; 3.716, 5.243; 3.853,4. 783 respectively, Ps ＜ 0. 05 ), while the CL and Oxygenation Index increased significantly ( t = 3. 976,4. 267; 4. 557,4. 265 respectively, P ＜ 0. 05 ). The duration of mechanical ventilation follow operation in intraaortic injection group ( [8. 03 ± 5. 14] h ) was shorter compared with intra-venous injection group ( [10. 56 ±6.95]h) (t =2.599,P＜0.05). Conclusion By intra-aortic protamine injection the lung injury decreased significantly. It shows good protective effect on lung in infants underwent opening heart operation by cardiopulmonary bypass surgery.

Objective To study the changes of behavior and depression-like classic indicators after hippocampal microinjection of K252a,and to establish a new animal model of depression.Methods SD rats were randomly divided into five groups,namely the control group,sham group,chronic stress depression model group,hippocampal of K252a microinjection group,and hippocampal microinjection K252a plus chronic stress group.Open field experiments,sucrose consumption test,and Morris water maze behavioral assay were used to assess the behavioral changes in the rats.ELISA was used to detect the plasma monoamine neurotransmitter,radioimmunoassay was used to determine the plasma CRH,ACTH,CORT contents,and western-blotting was performed to observe the protein expression of BDNF,CREB,ERK1/2,and BCL-2 in the hippocampus.Results Compared with the control group,the amount of activity,sugar consumption,learning and memory abilities were decreased(P<0.05 or P<0.01),also the serum monoamine neurotransmitters were decreased (P<0.01),HPA axis function was improved (P<0.01),and the expression of BDNF,CREB,ERK1/2,BCL-2 decreased in the CUMS group(P<0.05 or P<0.01),but there was no significant difference in the DMSO group.Compared with the DMSO group,the activity,consumption of sucrose,learning and memory ability were significantly decreased(P<0.05 or P<0.01),while the HPA axis function was increased (P<0.05 or P<0.01),the serum monoamine neurotransmitters decreased(P<0.05 or P<0.01),and the BDNF,CREB,ERK1/2,BCL-2 expressions in the hypocampus were significantly decreased(P<0.05 or P<0.01) in the K252a group and K252a + CUMS group.Compared with the CUMS group,the K252a group and K252a + CUMS group did not show significant changes in these parameters.Compared with the K252a group,these indicators were not significantly changed in the K252a + CUMS group.Conclusions The results of behavior,hematology,and molecular biology analysis show that this model has a great similarity to the classical model of CUMS in surface validity,construct validity,and functional validity.It may provide an alternative investigative technology platform for basic research and antidepressant drug screening.