Objective To explore the effect of erythropoietin (EPO) on cognition and neuron apoptosis in CA1 region of hippocampus in vascular dementia (VaD) rats.Methods 54 Wistar rats were randomly divided into three groups:sham control group,VaD group,VaD+ EPO group (n=18,each).The bilateral common carotid arteries of Wistar rats were permanently ligated to establish VaD models.Spatial study and memory were observed by Y maze test at 4,8 and 12 weeks after operation.Neuron apoptosis in CA1 area of hippocampus was measured by terminal deoxynucleotidyl-transferase (TdT) mediated dUTP-biotin nick end labeling (TUNEL) method.The protein and mRNA expressions of Bcl 2 and Bax in CA1 region of hippocampus were detected by immunohistochemistry and RT-PCR at 4,8,12 weeks after operation.Results Compared with VaD group,the VaD+ EPO group had better performances including less error reaction times and shorter total reaction time in Y-maze (both P＜0.05) at 4,8,12 weeks after operation.The apoptotic neuronal cells in CA1 region of hippocampus was decreased in VaD+ EPO group than in VaD group at 4,8,12 weeks after operation [(10.50±±2.43) vs.(20.50±± 3.29),(23.92±±3.18) vs.(33.58±3.48) and (36.92±4.10) vs.(54.17±4.26),t=4.23,3.54,5.05,P=0.013,0.024,0.007,respectively].The Bcl-2-positive cells and Bcl-2 mRNA expression were increased,and the Baxpositive cells and Bax mRNA expression were decreased in VaD+ EPO group than in VaD group (all P ＜0.05).Conclusions Erythropoietin can improve cognitive function by inhibiting neuron apoptosis through regulation of Bcl-2 and Bax expression in CA1 region of hippocampus.

Purpose　The aim is to find out an effective preventing influenza immune preparation. Methods　Egg yolk immune solution of anti-influenza virus(FM1 stem) was prepared from egg of the hens that had been successfully immunized with influenza virus (FM1 stem). Its effects of anti-virus were observed through animal experiment.Results　When the mice of control group began to die the average daily drink quantity of the mice of normal saline control group, egg yolk immune solution control group, immediate preventive group was (3.06±0.86), (2.93±1.47) and (3.99±0.21)ml(P＜0.05)respectively. The average body weights of the mice of these three groups were (15.85±2.70),(14.58±1.92) and (18.27±1.71)g(P＜0.05)respectively. Their mortality was separately 53.84%,69.23% and 3.84%(P＜0.01). The antibody positive rate of survived mice′s serum (1∶10 diluted) was separately 100%, 100% and 0%(P＜0.01). Conclusion　The anti-virus effects of egg yolk immune solution of anti-influenza virus was powerful. The result of preventing mice′s influenza was remarkable.

Objective To observe the effects of transdermal acupoint electric stimulation(TEAS)assisting sodium nitroprus-side induced controlled hypotension on serum glucose (Glu),angiotensin Ⅱ (ATⅡ)and superoxide dismutase (SOD),and to inves-tigate the protective effect of TEAS under controlled hypotension anesthesia.Methods 60 cases undergoing elective endoscopy si-nus surgery by adopting sodium nitroprusside induced controlled hypotension under general anaesthesia maintained the mean arterial pressure(MAP)in 50-60 mm Hg and were randomly and equally divided into two groups.The group Ⅰ conducted TEAS,while the group Ⅱ did not conduct TEAS.The controlled hypotension time and surgery time were recorded in the two groups;Glu and ATⅡ values were detected before anesthesia (T0 ),30 min after hypotension (T1 ),hypotension stopping(T2 );SOD was detected at T0 ,T2 ,30 min after hypotension(T3 ).Results The operation time and controlled hypotension continuous time had no statistically significant difference between the two groups(P >0.05).The Glu value in the group Ⅰ had no statistically significant difference a-mong the 3 time points,while which at T1 ,T2 was higher than that at T0 in the group Ⅱ(P <0.05),and which at T1 ,T2 in the group Ⅱ was higher than that in the group Ⅰ (P <0.05);the ATⅡ value at T1 was higher than that at T0 in the group Ⅰ (P <0.05),while which at T1 ,T2 was higher than that at T0 in the group Ⅱ(P >0.05),which at T1 ,T2 in the group Ⅱwas higher than that in the group Ⅰ (P <0.05);the SOD value at T2 was lower than that at T0 in the group Ⅰ,which at T2 ,T3 in the group Ⅱwas lower than that in the group Ⅰ(P <0.05).Conclusion TEAS assisting sodium nitroprusside controlled hypotension can better in-hibit the stress response.

Objective To investigate the effect of interleukin-1β (IL-1β) on the expression of synaptic protein SNAP-25 in the hippocampus in septic neonatal rat induced by systemic lipopolysaceharide (LPS) injection.Methods Sprague-Dawley (SD) rats were randomly divided into two groups:control group and sepsis group.The rat model of sepsis was produced by intraperitoneal injection of 1 mg/kg LPS,and rats in the control group were injected with an equal volume of 0.01 mol/L phosphate buffered saline (PBS).The expression levels of IL-1β and IL-1R1 in the hippocampus at 1,2 and 3 d,and synaptosomal-associated protein 25 (SNAP-25) at 7,14 and 24 d after LPS intraperitoneal injection were detected by Western blot.After cultured for 24 h,primary hippocampal neurons were divided into four groups including the control group,IL-1β (40 ng/mL) treatment group,IL-1β (40 ng/mL) + IL-1Ra (40 ng/mL) treatment group,and IL-1Ra (40 ng/mL) treatment group.The effect of IL-1β on SNAP-25 expression in primary hippocampal neuron was determined by Western blot and real-time PCR.The purity of hippocampal neurons were identified by NeuN immunofluorescence staining and the activity of neurons were detected by CCK-8 assay.All data were analyzed by SPSS version 22.0.The data were analyzed by student-t test and Dunnett-t test.The interaction effects were analyzed by factorial ANOVA.Differences were considered to be statistically significant if P＜ 0.05.Results Compared with the control group,the expressions of IL-1β and IL-1R1 were significantly increased in the hippocampus at 1,2 and 3 d after intraperitoneal injection of LPS (P＜0.05).The expression of SNAP-25 protein was decreased at 7,14,and 28 d after intraperitoneal injection of LPS (P＜0.05).The purity of primary neurons was about up to 92％.The activity of primary neurons was not relatively changed after treated with IL-1β at a dose less than 40 ng/mL.The level of SNAP-25 protein was obviously decreased in primary neurons at 24 h after IL-1β treatment (P＜0.05).IL-1Ra treatment might reverse the effect of IL-1β on primary neurons (P＜0.05).While,the expression of SNAP-25 mRNA was not statistically different in each group (P＞0.05).Conclusions IL-1β may possibly inhibit the expression level of SNAP-25 protein in the hippocampus in the septic rats through its receptor IL-1R1,which would contribute to cognitive dysfunction of septic neonatal rats in later life.

Objective:To analyze prognostic factors of hepatocellular carcinoma (HCC) combined with portal vein tumor thrombus (PVTT) after surgical resection.Methods:The data of 98 patients with HCC combined with PVTT who underwent surgical resection in Ningbo University Affiliated Li Huili Hospital from January 2008 to June 2019 were analyzed retrospectively, including 83 males and 15 females with an average age of 53 years. The survival rate was calculated by Kaplan-Meier method and compared using log-rank test. Cox regression model was used for the multivariate analysis of the prognosis of patients.Results:The 1, 2, and 3-year overall survival rates were 75.9%, 51.5%, and 35.4%, respectively, with a median survival time of 25 months; the 1, 2, and 3-year disease-free survival rates were 35.0%, 16.1%, and 8.6%, respectively, with a median disease-free survival time of 8 months. Multivariate analysis showed that the overall survival of patients with preoperative alpha-fetoprotein (AFP) ≥400 μg/L ( HR=1.760, 95% CI: 1.079-2.873) and hepatic vein tumor thrombus (HVTT, HR=3.809, 95% CI: 1.655-8.765) was poorer after surgical resection (all P<0.05), while the survival of patients with postoperative adjuvant trans-arterial chemoembolization (TACE) ( HR=0.397, 95% CI: 0.220-0.716, P=0.002) was better. Preoperative AFP≥400 μg/L ( HR=2.339, 95% CI: 1.488-3.676) , undergoing HCC resection combined with PVTT dissection ( HR=2.038, 95% CI: 1.090-3.811), and with HVTT ( HR=2.374, 95% CI: 1.160-4.857) (all P<0.05) are independent risk factors for recurrence in patients with HCC combined with PVTT, postoperative adjuvant TACE ( HR=0.535, 95% CI: 0.307-0.933, P=0.027) is a protective factor. Conclusion:Preoperative AFP≥400 μg/L and HVTT are independent risk factors for the prognosis of patients with HCC combined with PVTT. Reasonable selection of surgical methods and postoperative adjuvant TACE may improve the prognosis of patients.

Objective:To investigate the effect of melatonin on oligodendrocyte maturation and differentiation in corpus callosum of septic neonatal rats induced by systemic lipopolysaccharide (LPS) injection.Methods:Sprague-Dawley rats were randomly allocated into the control group, septic experimental group, and melatonin group. In the septic experimental group, rats were intraperitoneally administrated with lipopolysaccharide (LPS) (1 mg/kg). In the melatonin group, melatonin was intraperitoneally administered (10 mg/kg) at 0.5 h after LPS injection. The expression level of IL-6, olig1, olig2, and the MAG protein were detected by Western blot at different time points in the three groups. BV-2 cells were used in vitro. For drug administration, the effect of LPS, melatonin and melatonin receptor antagonist, luzindole, on IL-6 expression in BV-2 microglia cell was determined by Western blot. The medium of BV2 cell were collected to treat primary OPCs. The expression level of olig1, olig2 and MAG protein in primary OPCs were detected by Western blot. SPSS 20.0 statistical software was used for analysis, and the data were analyzed by one-way ANOVA and two-way ANOVA. Differences were considered to be statistically significantly if P<0.05. Result:Compared with the LPS group, the expression of IL-6 was significantly decreased in the corpus callosum at 6 h, l d, and 3 d in the melatonin group ( P<0.05). The expression of olig1, olig2 and MAG protein were increased at day 7, 14, and 28 in the melatonin group compared with the LPS group ( P<0.05). In vitro the expressions of IL-6 was significantly increased after LPS treatment ( P<0.05), but was decreased in the LPS+melatonin treatment group ( P<0.05). After treatment with melatonin receptor inhibitor, luzindole, the expressions level of IL-6 was increased ( P<0.05). The expression of olig1, olig2 and MAG protein were decreased with conditioned medium in the LPS BV2 cell group than the control group in the primary OPCs ( P<0.05). However, those were increased with conditioned medium in the LPS+melatonin BV2 cell group than the LPS group ( P<0.05). Conclusions:Melatonin may inhibit the inflammation response in the corpus callosum through its receptor, and may promote the maturation and differentiation of oligodendrocyte, suggesting that melatonin may have therapeutic effect on neuroinflammation and axonal hypomyelination on PWM in septic neonatal rats.

Peribiliary glands (PBG) is a kind of microscopic structure around the intra-hepatic bile ducts and extrahepatic bile ducts. PBG not only participates in maintaining the normal physiological function of biliary epithelial tissue, but also plays an important role in its damage and repair process. Biliary tree stem/progenitor cells in PBG are important cell sources of biliary epithelial regeneration and repair. PBG and the surrounding peribiliary vascular plexus are key influencing factors for the occurrence of ischemic-type biliary lesions (ITBL) after liver transplantation. Based on relevant literatures and clinical practice, the authors summarize the function of PBG as well as its relationship with ITBL.

Objective:To study the safety and efficacy of combining portal vein resection and reconstruction (PVR) with resection of perihilar cholangiocarcinoma (PHC).Methods:A total of 104 patients with PHC who underwent hepatectomies for either biliary resection alone or biliary resection combined with PVR from October 2006 to December 2019 at the Department of Hepatopancreatobiliary, Ningbo Medical Center of Lihuili Hospital entered into this study. There were 63 males and 41 females, with the age of (64.4±10.4) years. The control group consisted of 75 patients who underwent biliary resection alone, while the PVR group consisted 29 patients with biliary resection combined with PVR. The patient characteristics and the follow-up outcomes of the two groups were analyzed and compared. Survival analyses were performed using the Kaplan Meier method with the log-rank test.Results:Wedge resection of portal vein, side to side anastomosis in 2 cases, segmental resection and end to end anastomosis in 27 cases. The time taken for PVR and portal vein resection were (12.7±2.9)(range 8 to 18)min and (20.7±7.3)(range 8 to 38) mm, respectively. The estimated blood loss for the PVR group was significantly more than the control group [ M( Q1, Q3)] 800.0 (600.0, 1 500.0) ml vs. 600.0(500.0, 1 000.0) ml ( P<0.05). Based on postoperative pathological studies, the proportion of lymph node metastasis was significantly higher in the PVR group than the control group (58.6% vs. 32.0%, P<0.05). Clavien-Dindo grade Ⅲ and above complications were 30.7%(23/75) and 34.5%(10/29) in the control and PVR groups, respectively ( P>0.05). The re-operation and postoperative 90 days mortality rates were 9.3%(7/75) and 2.7%(2/75) in the control group, compared with 3.4%(1/29) and 0 in the PVR group, respectively (both P>0.05). The 1-, 3- and 5-year survival rates were 81.1%, 44.8% and 36.4% respectively for the control group and 78.1%, 35.9% and 31.4% for the PVR group (χ 2=0.33, P=0.570). Conclusion:When compared to biliary resection alone, biliary resection combined with PVR did not significantly increase postoperative complication and mortality rates, but with comparable long-term survival outcomes. Combined biliary resection with PVR was safe and improved the resection rate in selected patients with locally advanced PHC.

Objective:To evaluate the short-term efficacy of split liver transplantation (SLT) in patients with acute-on-chronic liver failure (ACLF).Methods:The clinical data of 9 ACLF patients receiving SLT in our center from Mar 2021 to May 2022 were retrospectively analyzed to evaluate its safety and efficacy.Results:The preoperative APASL ACLF Research consortium (AARC) score of the 9 ACLF patients was 8 points in 1 case, 9 points in 3 cases, 10 points in 3 cases, 11 points in 1 case and 12 points in 1 case, 7 cases were in AARC-ACLF grade 2, and 2 cases in grade 3.In-situ liver splitting was performed in 9 deceased donors, including 4 classical split cases, 5 full size split cases. Among these 9 ACLF patients, 2 received left half liver transplantation, 3 received right half liver transplantation, and 4 received extended right lobe liver transplantation. After transplantation, all 9 recipients were discharged fully recovered, 1 case developed Clavien grade Ⅳa complication and 2 cases developed Clavien grade Ⅲb complication.After SLT treatment the median postoperative hospital stay was 27 days, the 1-year survival rate was 100%, and the organ survival rate was 88.9%.Conclusion:Split liver transplantation is a safe and feasible treatment method for ACLF patients.

Objective:To explore the feasibility of technological modification and innovation of full-left/full-right liver splitting in situ for donors and examine the safety of clinical application for liver transplantation (LT).Methods:From March 2021 to June 2022, clinical and surgical data are retrospectively reviewed for 27 donors undergoing full-left/full-right liver splitting in situ and the corresponding 49 recipients undergoing full-left/full-right LT.According to the split liver technique used in donor liver surgery, they are divided into conventional split group(group A, 13 cases)and innovative split group(group B, 14 cases). The corresponding recipients are divided into two groups of recipient C(25 cases)and recipient D(24 cases). General profiles, intraoperative findings, type of vascular allocation and short-term outcomes in two groups are compared.After full-size split liver transplantation(fSLT), follow-ups continued until the end of September 2022.Results:There are 23 males and 4 females in donors.The causes of mortality for donors are traumatic head injury(12 cases)cerebrovascular accident(13 cases)and anoxia encephalopathy(2 cases). Baseline characteristics of two groups indicate that body weight and body mass index(BMI)are higher in group B and blood sodium level is lower than that in group A( P<0.05). No statistical differences exist for the others.Liver splitting time is significantly shorter in group B than that in group A(175 vs.230 min, P=0.022). No significant inter-group difference exists in type of vascular allocation.Retrohepatic inferior vena cava(IVC)is split in one case in group A and 10 cases in group B( P=0.001). Among 20 cases of right hemiliver requiring a reconstruction of segment Ⅴ/Ⅷ venous outflow, 12 cases in group A and 3 cases in group B are reconstructed with conventional independent bridging method(independent type)while another 5 cases in group B reconstruct with innovated technique by bridging Ⅴ/Ⅷ vein for splitting IVC with iliac vessel and molding all outflows as one for anastomosis(combined typ e). There is significant inter-group difference( P=0.004). No significant differences exist in operative duration, anhepatic phase or blood loss between groups C and B, except for T tube retaining in 7 cases of group A and 14 cases of group D( P=0.032). Twelve cases developed a total of 26 instances of≥Clavien-Dindo grade Ⅲ complications.Of which, 7 cases in group C and 5 cases in group D show no significant difference in postoperative morbidity.However, for serious biliary complications(≥Clavien Dindo grade Ⅲ), there are 6 cases in group C versus none in group D( P=0.016). Two cases died from postoperative complication with a postoperative mortality rate of 4.1%.Postoperative hospital stay is similar in two groups.And accumulates 6/12-month survivals were 95.9% and 87.7% for grafts and 95.9% and 92.4% for recipients respectively. Conclusions:Operative duration of full-left/full-right liver splitting in situ tends to shorten with an accumulation of a certain amount of cases.Technological modification and innovation in IVC splitting and segment Ⅴ/Ⅷ vein reconstruction should be further validated as both feasible and safe by short-term outcomes of the corresponding recipients.