Objective To explore the effects of prucalopride(PRUC)on the intestinal function during the perioperative period of robot-assisted laparoscopic radical cystectomy(RARC)and urinary diversion.Methods A total of 75 patients undertaking RARC with urinary diversion(orthotopic neobladder or ileal bladder)in Nanjing Drum Hospital during Jan.and Dec.2021 were divided into PRUC group(n=28)and control group(n=47)according to whether they took PRUC or not.Postoperative intestinal ventilation time and defecation time,drainage tube retention time,tolerance time for first intake of semi-flow food,postoperative hospital stay,and incidence of complications were observed and recorded in the two groups.Postoperative C-reactive protein(CRP)and neutrophil/lymphocyte ratio(NLR)were compared.Results The PRUC group had shorter intestinal ventilation time and defecation time[(47.14±16.31)h vs.(74.04±35.33)h,P＜0.01;(86.14±30.47)h vs.(123.57±79.12)h,P=0.02],smaller change of ΔCRP and ΔNLR[(79.99±29.71)mg/L vs.(127.75±56.98)mg/L;(9.24±6.43)vs.(16.11±9.90),P＜0.01].All complications were minor,the incidence of intestinal obstruction in PRUC group tended to decrease within 90 days after operation(P=0.38),and there was no significant difference in other complications between the two groups(P＞0.05).Conclusion The perioperative use of PRUC in RARC with urinary diversion is safe and effective,which can promote the recovery of intestinal function after operation.

Objective To explore the value of transesophageal echocardiography(TEE)for surgical therapy of pediatric atrioventricular valve diseases.Methods Data of 273 children with atrioventricular valve diseases who underwent surgical operation under extracorporeal circulation were retrospectively analyzed.Transthoracic echocardiography(TTE)was performed within 1 week before,while TEE was performed before and after surgical operation.Whether TTE diagnosis should be adjusted were evaluated according to findings of TEE,so were the effects of surgical treatments,and the ultrasonic diagnostic coincidence rate before operation were compared taken surgical findings as standards.Results TEE was successfully completed in all 273 children.The diagnostic coincidence rate of preoperative TTE was 83.88％(229/273).After adjustment or supplement according to TEE in 44 cases(44/273,16.12％),the ultrasonic diagnostic coincidence rate reached 100％(273/273),higher than that of preoperative TTE alone(P＜0.001).Residual shunt was found in 5 cases,while no obvious improvement of valvular regurgitation was noticed in 3 cases with post operative TEE,and after corresponding treatments,re-examination of TEE showed satisfactory efficacy in all the above cases.Conclusion TEE could be used to accurately diagnose pediatric atrioventricular valve diseases before and evaluate the efficacy after surgical treatments,having important clinical application value.

Background Paraquat (PQ), one of the environmental poisons associated with sporadic Parkinson's disease (PD), can cause abnormal aggregation of alpha-synuclein (α-syn), but the research on its conformational changes and subcellular localization is limited. Objective To investigate the effect of PQ on α-syn conformation and subcellular localization in dopaminergic neurons. Methods Forty-eight SPF C57BL/6 male mice were selected and randomly divided into a control group and a model group. The model group was intraperitoneally injected with PQ (15 mg·kg⁻¹), and the control group was intraperitoneally injected with 0.9% normal saline, twice a week for eight weeks to construct a PD-like mouse model. The changes of neurobehavior (by open field test and pole climbing test) were observed to evaluate motor ability of mice. Immunohistochemical staining (IHC) was used to detect the expression levels of tyrosine hydroxylase (TH) and α-syn in the midbrain. Western blotting (WB) was used to measure the protein expression levels of TH and α-syn in midbrain. Human neuroblastoma SH-SY5Y cells were used as dopaminergic neuron in vitro models. After the cells were treated with PQ (100 μmol·L⁻¹) for 0, 12, 24, 36 and 48 h, the expressions of α-syn in whole cell, cytoplasm, and nucleus were detected by WB; the expression level of extracellular α-syn was detected by enzyme-linked immunosorbent assay (ELISA); the change of α-syn location was observed by immunofluorescence assay (IFA). Results The neurobehavioral tests' results showed that compared with the control group, the residence time in peripheral area of mice in the PQ model group increased with the increase of exposure time (P<0.05), the residence time and moving distance in the central region decreased (P<0.05), and the pole climbing time increased (P<0.05). The mouse IHC results showed that compared with the control group, the number of TH positive cells in the midbrain decreased in the model group at week 6 and 8 (P<0.05), while the expression level of α-syn increased at week 4, 6, and 8 (P<0.05). The WB results of mouse showed that the relative expression of TH decreased significantly after 6 and 8 weeks of PQ exposure (P<0.05), and the relative expression of oligomer α-syn increased after 4, 6, and 8 weeks of PQ exposure (P<0.05). The WB of in vitro models results showed that the relative expression of α-syn in cells increased with time (R²=0.7440, P<0.05); the relative expression of α-syn in cytoplasm increased firstly and then decreased with time (P<0.05); the relative expression of α-syn in nucleus increased with time (R²=0.7913, P<0.05). The IFA results of in vitro models showed that the expression of oligomerized α-syn increased and translocated to the nucleus (P<0.05). The ELISA results of in vitro models showed that α-syn increased with the increase of PQ exposure time (P<0.05). Conclusion PQ can increase the expression of α-syn in dopaminergic neurons, induce oligomerization and translocation to the nucleus.

Objective:To explore the mechanism of HDAC6 mediated aggresome-autophagy-lysosome pathway in paraquat-induced autophagy in dopaminergic neurons.Methods:Human neuroblastoma cell (SH-SY5Y cell) was used as model of dopaminergic neurons in vitro. The cells were treated with terminal concentrations of 0, 25, 50, 100, 200 and 400μmol/L PQ for 24 hours, and the cells were induced by 100 μmol/L PQ for different time (0, 12, 24, 36, 48, 60 and 72 h) . Cell viability was detected by CCK-8 assay. The expression levels of HDAC6, α-syn, Dynein IC1/2, LC3, Beclin1, p62 and Lamp-1 were detected by Western blot. Immunofluorescence double-labeling method was used to observe the expression and localization of HDAC6, α-syn, Dynein IC1/2, LC3, Lamp-1 and γ-tubulin in cells.Results:CCK-8 assay showed PQ induced cell survival rate decrease in a time and dose dependent manner ( R=-0.950、-0.960, P<0.05) .Western blot showed that compared with control group, the protein levels of HDAC6, α-syn, p62 in PQ-exposed group were significantly increased ( P<0.05) , but there was a significant decrease in expression level of the ratio of autophagy-related protein LC3 Ⅱ/LC3 Ⅰ, Beclin1, Dynein IC1/2, Lamp-1in PQ-exposed group ( P<0.05) . The results of immunofluorescence double-labeling showed that compared with the control group, the fluorescence signals of HDAC6 and α-syn in the PQ-exposed group increased, and the protein expression level increased, while the fluorescence signals of Dynein IC1/2, LC3, and Lamp-1 decreased. The protein expression level is reduced. HDAC6 gradually accumulates from the diffuse shape to the nucleus; Under normal circumstances, α-syn, Dynein IC1/2, γ-tubulin, LC3, and Lamp-1 are mainly distributed in the cytoplasm. After PQ is infected, they gather in the nucleus and co-localize with HDAC6 in the area around the nucleus. Conclusion:PQ may induce abnormal aggregation of α-syn by inducing HDAC6-mediated aggresome-autophagy-lysosomal pathway disorder.

Objective:To explore the mechanism of HDAC6 mediated aggresome-autophagy-lysosome pathway in paraquat-induced autophagy in dopaminergic neurons.Methods:Human neuroblastoma cell (SH-SY5Y cell) was used as model of dopaminergic neurons in vitro. The cells were treated with terminal concentrations of 0, 25, 50, 100, 200 and 400μmol/L PQ for 24 hours, and the cells were induced by 100 μmol/L PQ for different time (0, 12, 24, 36, 48, 60 and 72 h) . Cell viability was detected by CCK-8 assay. The expression levels of HDAC6, α-syn, Dynein IC1/2, LC3, Beclin1, p62 and Lamp-1 were detected by Western blot. Immunofluorescence double-labeling method was used to observe the expression and localization of HDAC6, α-syn, Dynein IC1/2, LC3, Lamp-1 and γ-tubulin in cells.Results:CCK-8 assay showed PQ induced cell survival rate decrease in a time and dose dependent manner ( R=-0.950、-0.960, P<0.05) .Western blot showed that compared with control group, the protein levels of HDAC6, α-syn, p62 in PQ-exposed group were significantly increased ( P<0.05) , but there was a significant decrease in expression level of the ratio of autophagy-related protein LC3 Ⅱ/LC3 Ⅰ, Beclin1, Dynein IC1/2, Lamp-1in PQ-exposed group ( P<0.05) . The results of immunofluorescence double-labeling showed that compared with the control group, the fluorescence signals of HDAC6 and α-syn in the PQ-exposed group increased, and the protein expression level increased, while the fluorescence signals of Dynein IC1/2, LC3, and Lamp-1 decreased. The protein expression level is reduced. HDAC6 gradually accumulates from the diffuse shape to the nucleus; Under normal circumstances, α-syn, Dynein IC1/2, γ-tubulin, LC3, and Lamp-1 are mainly distributed in the cytoplasm. After PQ is infected, they gather in the nucleus and co-localize with HDAC6 in the area around the nucleus. Conclusion:PQ may induce abnormal aggregation of α-syn by inducing HDAC6-mediated aggresome-autophagy-lysosomal pathway disorder.

Objective:To investigate the protective effect of taurine (Tau) on hippocampus, substantia nigra neurons and microglia in paraquat (PQ) -induced Pakinson's disease-like mice.Methods:In April 2019, the specific pathogen free (SPF) C57BL/6 mice ( n=36) were randomly divided into control group (NaCl) , Tau control group (150 mg/kg) , PQ exposure group (10 mg/kg PQ group, 15 mg/kg PQ group) , Tau intervention group (Tau+10 mg/kg PQ group, Tau+15 mg/kg PQ group) , respectively. Tau was used in 1 h before PQ administration for consecutive 6 weeks (twice per week) . General and neurobehavioral tests (Traction test, Open field test, Forced Swimming test, Tail suspension test, High plus maze and Object recognition test) were performed to test motor and cognitive function. After neuroethology detection, mice were euthanized and brains were collected. Nissl staining was used to detect the changes of the number and morphology of Nissl bodies in hippocampus and substantia nigra neurons of mice. Immunohistochemistry (IHC) was used to test the levels of neuron marker neuronal nuclei antigen (NeuN) , substantia nigra dopaminergic neuron marker tyrosine hydroxylase (TH) , α-synuclein (α-syn) , microglia markers ionized calcium bindingadaptor molecule-1 (Iba-1) , inducible nitric oxide synthase (iNOS) and interleukin-1β (IL-1β) in mice substantia nigra. The coexpression of Iba-1 and TH double-labeling, α-syn and TH double-labeling in mice substantia nigra were measured by immunofluorescence double staining. Results:General behavioral changes such as slow reaction and reduced action occurred in mice of PQ group. Compared with the control group, the scores of Traction test, and the time ratio of new object recognition in the PQ group decreased ( P<0.05) , the fixed time of Swimming test and Tail suspension test increased ( P<0.05) , the horizontal crawl number and vertical times of Open field test and the ratio of open arm residence time of High plus maze in the 15 mg/kg PQ group decreased ( P<0.05) . Compared with the PQ group, the same dose of Tau+PQ group showed increased scores in Traction test ( P<0.05) and decreased fixed time of Swimming test and Tail suspension test ( P<0.05) . Compared with the 15 mg/kg PQ group, the horizontal crawl number of Open field test and the time ratio of new object recognition increased in the Tau+15 mg/kg PQ group ( P<0.05) . Compared with the control group, the PQ group showed a decrease in the number of Nissl body in the hippocampus and substantia nigra ( P<0.05) , a decrease in the number of NeuN and TH positive cells in the substantia nigra ( P<0.05) , with a large number of α-syn deposition, Iba-1 activation of microglia cells, and an increase in the expression of inflammatory factors (IL-1β, iNOS) in the hippocampus and substantia nigra ( P<0.05) . Compared with the PQ group, the same dose of Tau+PQ group showed the number of Nissl in the hippocampus and substantia nigra was significantly increased ( P<0.05) , the number of NeuN and TH positive cells in the substantia nigra was significantly increased ( P<0.05) , the expression levels of α-syn, Iba-1 and inflammatory factors (IL-1β, iNOS) in the substantia nigra were significantly decreased ( P<0.05) . Conclusion:Tau could protect PQ-induced degeneration of substantia nigra dopaminergic neurons and hippocampal neuron loss by inhibiting the activation of microglia cells and release of inflammatory factors, and effectively improve the neurobehavioral and brain histopathological changes of PQ-induced PD-like mice.

Objective:To investigate the protective effect of taurine (Tau) on hippocampus, substantia nigra neurons and microglia in paraquat (PQ) -induced Pakinson's disease-like mice.Methods:In April 2019, the specific pathogen free (SPF) C57BL/6 mice ( n=36) were randomly divided into control group (NaCl) , Tau control group (150 mg/kg) , PQ exposure group (10 mg/kg PQ group, 15 mg/kg PQ group) , Tau intervention group (Tau+10 mg/kg PQ group, Tau+15 mg/kg PQ group) , respectively. Tau was used in 1 h before PQ administration for consecutive 6 weeks (twice per week) . General and neurobehavioral tests (Traction test, Open field test, Forced Swimming test, Tail suspension test, High plus maze and Object recognition test) were performed to test motor and cognitive function. After neuroethology detection, mice were euthanized and brains were collected. Nissl staining was used to detect the changes of the number and morphology of Nissl bodies in hippocampus and substantia nigra neurons of mice. Immunohistochemistry (IHC) was used to test the levels of neuron marker neuronal nuclei antigen (NeuN) , substantia nigra dopaminergic neuron marker tyrosine hydroxylase (TH) , α-synuclein (α-syn) , microglia markers ionized calcium bindingadaptor molecule-1 (Iba-1) , inducible nitric oxide synthase (iNOS) and interleukin-1β (IL-1β) in mice substantia nigra. The coexpression of Iba-1 and TH double-labeling, α-syn and TH double-labeling in mice substantia nigra were measured by immunofluorescence double staining. Results:General behavioral changes such as slow reaction and reduced action occurred in mice of PQ group. Compared with the control group, the scores of Traction test, and the time ratio of new object recognition in the PQ group decreased ( P<0.05) , the fixed time of Swimming test and Tail suspension test increased ( P<0.05) , the horizontal crawl number and vertical times of Open field test and the ratio of open arm residence time of High plus maze in the 15 mg/kg PQ group decreased ( P<0.05) . Compared with the PQ group, the same dose of Tau+PQ group showed increased scores in Traction test ( P<0.05) and decreased fixed time of Swimming test and Tail suspension test ( P<0.05) . Compared with the 15 mg/kg PQ group, the horizontal crawl number of Open field test and the time ratio of new object recognition increased in the Tau+15 mg/kg PQ group ( P<0.05) . Compared with the control group, the PQ group showed a decrease in the number of Nissl body in the hippocampus and substantia nigra ( P<0.05) , a decrease in the number of NeuN and TH positive cells in the substantia nigra ( P<0.05) , with a large number of α-syn deposition, Iba-1 activation of microglia cells, and an increase in the expression of inflammatory factors (IL-1β, iNOS) in the hippocampus and substantia nigra ( P<0.05) . Compared with the PQ group, the same dose of Tau+PQ group showed the number of Nissl in the hippocampus and substantia nigra was significantly increased ( P<0.05) , the number of NeuN and TH positive cells in the substantia nigra was significantly increased ( P<0.05) , the expression levels of α-syn, Iba-1 and inflammatory factors (IL-1β, iNOS) in the substantia nigra were significantly decreased ( P<0.05) . Conclusion:Tau could protect PQ-induced degeneration of substantia nigra dopaminergic neurons and hippocampal neuron loss by inhibiting the activation of microglia cells and release of inflammatory factors, and effectively improve the neurobehavioral and brain histopathological changes of PQ-induced PD-like mice.

Objective To explore the HO-1 expression levels with 5-FU chemosensitivity in esophageal squamous cell carcinoma.Methods Eca109 cells used in all experiments,MTT assay was used in cell growth curve and inhibit rate.RT-PCR and Western blot was used to detect HO-1 in cells treated with different concentrations of ZnppⅨ (0,20,80,100 μmol/L).Results The inhibitory rate of cells was significantly increased when the concentrition of ZnppⅨ increased.The inhibitory rate of cells in 80 μmol/L ZnppⅨ was higher than 20 μmol/L ZnppⅨ group(P＜0.05).The expression of HO-1 mRNA in each group was 0.50± 0.17,0.55±0.15,0.58 ± 0.09 and 0.55 ± 0.16,respectively,there was no significant difference between the two groups (P＞ 0.05).The expression of HO-1 was 0,85±0.07,0.63±0.11,0.43±0.12 and 0.25±0.10,respectively.The expression of HO-1 had significant difference (F=20.01,P＜0.01).Conclusion Eca109 cells inhibition rate positive correlated with ZnppⅨ concentrations,and ZnppⅨ were inhibited the expression of HO-1.not from gene level,but after the translation level.

Objective To summarize the experience of the ventriculoperitoneal shunt in treating children with methylmalonic aciduria combined with hydrocephalus,and to assess the clinical value.Methods From September 2012 to May 2016,a total of 12 patients with methylmalonic aciduria combined with hydrocephalus in Peking University First Hospital were enrolled,including 7 boys and 5 girls.All the 12 patients underwent ventriculoperitoneal shunt.Drug therapy was performed after surgery.The clinical manifestations and imaging findings were used as the basis for adjusting the pressure of the diverter valve appropriately.The clinical condition of patients were evaluated retrospectively.The patients' clinical symptoms,signs,imaging materials,surgical complications and postoperative prognosis were analyzed.Results All the cases were followed up for 3 to 36 months,no death case and no serious postoperative complications of hydrocephalus occurred.Clinical symptoms of intracranial hypertension were relieved or disappeared.The head circumference progressive enlargement stopped.The anterior fontanelle tension decreased significantly.Setting-sun sign of eyes disappeared.One out of 4 cases with convulsion and epilepsy was relieved after the operation.Seven cases of poor vision or vision loss,postoperative visual acuity were improved though not recover to normal eyesight.One case of the children with hearing loss,postoperative hearing recovered.During the follow-up period,the head CT showed that the ventricle was narrowed significantly,interstitial brain edema improved obviously.Conclusion Ventriculoperitoneal shunt is a effective method for treating children with methylmalonic aciduria combined with hydrocephalus,which is beneficial for patients with these diseases.

SUMMARY Hemophilia A is aninherited bleeding disorder, lack of coagulation factorⅧ( FⅧ) , and if combined with intracranial malignant tumor, the operation risk is very high. Department of Pediatric Sur-gery in Peking University First Hospital used coagulation factor replacement therapy, succeeded in the operation of 2 cases of intracranial malignant tumor with hemophilia A in children, with no abnormal bleeding events. The establishment of the multi subject cooperation group before operation, good preoper-ative preparation, enough alternative factors, and close postoperative monitoring, are the key to the suc-cessful treatment.