ObjectiveTo investigate the mechanism by which adrenoceptor alpha 2A (Adra2a) regulates lipopolysaccharide (LPS)-induced inflammation in primary hepatocytes from lipopolysaccharide-binding protein (LBP) knockout mice (Lbp^-/-). MethodsPrimary hepatocytes from C57BL/6J and Lbp^-/- mice were isolated using a two-step perfusion method. An in vitro inflammatory model was established by LPS stimulation, and an in vivo inflammatory mouse model was established by intraperitoneal injection of LPS. The in vitro experiments were grouped as follows: Control group, LPS group, BRL+LPS group, OE-NC+LPS group, and OE-Adra2a+LPS group. The Control group served as the blank control. The LPS group involved stimulating primary hepatocytes with LPS. The BRL+LPS group involved pretreating primary hepatocytes with BRL-44408 maleate followed by LPS stimulation. The OE-NC+LPS group involved transfecting primary hepatocytes with an empty vector followed by LPS stimulation. The OE-Adra2a+LPS group involved transfecting primary hepatocytes with a lentivirus overexpressing Adra2a, followed by LPS stimulation. The in vivo experimental groups were divided into Control', LPS', BRL+LPS', OE-NC+LPS', and OE-Adra2a+LPS' groups. The Control' group served as the blank control. The LPS' group received intraperitoneal injection of LPS. The BRL+LPS' group received intraperitoneal injection of BRL-44408 maleate for pretreatment, followed by LPS injection. The OE-NC+LPS' group received intraperitoneal injection of empty vector for pretreatment, followed by LPS injection. The OE-Adra2a+LPS' group received intraperitoneal injection of a lentivirus overexpressing Adra2a for pretreatment, followed by LPS injection. Cell viability after Adra2a inhibition and overexpression was assessed via the Cell Counting Kit-8 (CCK-8) assay. RT-qPCR measured changes in gene expression levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) after Adra2a inhibition and overexpression. Western blotting was performed to detect Adra2a protein expression and phosphorylation levels of extracellular signal-regulated kinase 1/2 (ERK1/2), p38 mitogen-activated protein kinase, and c-Jun N-terminal kinase (JNK) following LPS stimulation. ResultsIn vitro experiments revealed that LPS stimulation significantly decreased Adra2a protein expression in primary hepatocytes from C57BL/6J mice compared to the Control group (P<0.05), whereas it increased in primary hepatocytes from Lbp^-/- mice (P<0.001). Compared to the LPS group, the BRL+LPS group exhibited significantly increased cell viability (P<0.01), reduced TNF-α, IL-6, and IL-1β gene transcription levels (P<0.01, P<0.001, P<0.001), and decreased phosphorylation levels of MAPK signaling pathway-related proteins ERK1/2, p38, and JNK (P<0.01, P<0.001, P<0.001). Compared with the OE-NC+LPS group, the OE-Adra2a+LPS group showed significantly decreased cell viability (P<0.001), increased gene transcription levels of TNF-α, IL-6, and IL-1β genes (P<0.001, P<0.01, P<0.001), and elevated phosphorylation levels of MAPK signaling pathway-related proteins ERK1/2, p38, and JNK (P<0.001, P<0.01, P<0.001). In vivo experiments showed that, compared with the LPS' group, the BRL+LPS' group exhibited significantly reduced phosphorylation levels of MAPK signaling pathway-related proteins ERK1/2, p38, and JNK (P<0.001, P<0.01, P<0.01). In the OE-Adra2a+LPS' group, the phosphorylation levels of ERK1/2, p38, and JNK were significantly elevated compared to the OE-NC+LPS' group (P<0.01, P<0.001, P<0.01). ConclusionLPS stimulation can cause a significant increase in Adra2a protein expression in primary hepatocytes of Lbp^-/- mice. Adra2a protein can regulate the level of LPS-induced inflammation in primary hepatocytes of Lbp^-/- mice through the MAPK signaling pathway.

The purpose of this study is to systematically determine an optimal percentile cutoff in histogram analysis for calculating the mean parameters obtained from a nonGaussian continuous-time random-walk (CTRW) diffusion model for differentiating individual glioma grades. This retrospective study included 90 patients with histopathologically proven gliomas (42 grade II, 19 grade III, and 29 grade IV). We performed diffusion-weighted imaging using 17 b-values (0-4000 s/mm²) at 3T, and analyzed the images with the CTRW model to produce an anomalous diffusion coefficient (D m ) along with temporal (α) and spatial (β) diffusion heterogeneity parameters. Given the tumor ROIs, we created a histogram of each parameter; computed the P-values (using a Student’s t-test) for the statistical differences in the mean D m , α, or β for differentiating grade II vs. grade III gliomas and grade III vs. grade IV gliomas at different percentiles (1% to 100%); and selected the highest percentile with P < 0.05 as the optimal percentile. We used the mean parameter values calculated from the optimal percentile cut-offs to do a receiver operating characteristic (ROC) analysis based on individual parameters or their combinations.We compared the results with those obtained by averaging data over the entire region of interest (i.e., 100th percentile). We found the optimal percentiles for D m , α, and β to be 68%, 75%, and 100% for differentiating grade II vs. III and 58%, 19%, and 100% for differentiating grade III vs. IV gliomas, respectively. The optimal percentile cut-offs outperformed the entire-ROI-based analysis in sensitivity (0.761 vs. 0.690), specificity (0.578 vs. 0.526), accuracy (0.704 vs. 0.639), and AUC (0.671 vs.0.599) for grade II vs. III differentiations and in sensitivity (0.789 vs. 0.578) and AUC (0.637 vs. 0.620) for grade III vs. IV differentiations, respectively. Percentile-based histogram analysis, coupled with the multi-parametric approach enabled by the CTRW diffusion model using high b-values, can improve glioma grading.

Objective To explore the clinical efficacy of radical resection with individualized surgical approach for borderline resectable pancreatic head carcinoma.Methods The retrospective descriptive study was conducted.The clinicopathological data of 54 patients with borderline resectable pancreatic head carcinoma who underwent radical resection with individualized surgical approach in the West China Hospital of Sichuan University from January 2015 to January 2018 were collected.There were 37 males and 17 females,aged from 37 to 73 years,with a median age of 59 years.For venous type borderline resectable pancreatic head carcinoma,surgery for pancreatic head carcinoma and (or) pancreatic head and neck carcinoma was performed via inferior mesenteric vein,and surgery for pancreatic uncinate process carcinoma was performed via inferior colon artery.For arterial type borderline resectable pancreatic head carcinoma,surgery for pancreatic head carcinoma and (or) pancreatic head and neck carcinoma was performed via medial uncinate artery,and surgery for pancreatic uncinate process carcinoma was performed via left posterior artery.Observation indicators:(1) surgical situations;(2) postoperative complications;(3) postoperative pathological examination;(4) follow-up.Patients were followed up by outpatient examination or telephone interview once every 3 months to detect survival up to March 2019.Measurement data with normal distribution were represented by Mean ± SD.Measurement data with skewed distribution were represented by M (range),and count data were represented by absolute numbers or percentage.Kaplan-meier method was used to draw the survival curve and calculate the survival rate.Results (1) Surgical situations:all the 54 patients underwent expanded pancreatoduodenectomy combined with superior mesenteric vein/portal vein (SMV/PV) resection,including 15 via inferior mesenteric vein,20 via inferior colon artery,12 via medial uncinate artery,and 7 via left posterior artery.The operation time was (320± 83)minutes,and the volume of intraoperative blood loss was (865±512) mL.(2) Postoperative complications:of 54 cases,28 had postoperative complications,including 13 with grade 1 Clavien-Dindo complications,12 with grade 2 ClavienDindo complications,3 with grade 3 or above Clavien-Dindo complications.One of the 28 patients with postoperative complications died and 27 were improved after symptomatic and supportive treatment.(3) Postoperative pathological examination:of 54 patients,31 had R0 resection and 23 had R1 resection.In the 23 patients with R1 resection,5 underwent surgery via the inferior mesenteric vein (4 with involvement of pancreatic anterior surface,1 with involvement of both pancreatic anterior and posterior surface),9 underwent surgery via the inferior colon artery (2 with involvement of both pancreatic anterior and posterior surface,2 with involvement of superior mesenteric artery margin,2 with involvement of pancreatic posterior surface,2 with involvement of pancreatic anterior surface,1 with involvement of superior mesenteric artery margin and pancreatic posterior surface),5 underwent surgery via the medial uncinate process artery (2 with involvement of superior mesenteric artery margin,2 with involvement of both pancreatic anterior and posterior surface,1 with involvement of pancreatic neck transected margin),and 4 underwent surgery via the left posterior artery (3 with involvement of superior mesenteric artery margin,1 with involvement of both pancreatic anterior and posterior surface).Of 54 patients,16 had no positive lymph nodes,26 had 1-3 positive lymph nodes,and 12 had 4 or more positive lymph nodes.The tumor diameter was (3.20±0.14)cm.There were 48 of 54 patients with nerve infiltration,41 with superior mesenteric vein and/or portal vein infiltration,and 11 with vascular thrombus.There were 17 of 54 patients with high differentiation and medium differentiation,and 37 with low differentiation and undifferentiation.(4) Follow-up:54 patients were followed up for 1-42 months,with a median time of 19 months.The 1-,3-year overall survival rate was 78.0％,11.4％.Condusion As for the borderline resectable pancreatic head cancer,individualized and customized surgical approach according to the location of tumor and the relationship with blood vessels is helpful to standardize the radical resection and avoid R2 resection.

OBJECTIVE To elucidate the toxicological properties of CdTe quantum dots (CdTe QD) by investigating their effect on mitophagy in human umbilical vein endothelial cells (HUVECs).METHODS The purity of primarily cultured HUVECs was detected by immunofluorescence.Then,HUVECs were incubated with CdTe QD 0.1-100 mg· L-1 for 24 h.After treatment,the cell viability of HUVECs was detected with MT-T assay.The mitochondrial morphology was observed under a laser scanning confocal microscope after labeling with Mitotracker.The treated HUVECs were also labeled with JC-1 probe,and the mitochondrial membrane potential (MMP) was then examined by flow cytometry.The expression of mitophagy-related proteins including microtubule-associated protein 1 light chain 3 Ⅰ /Ⅱ (LC3 Ⅰ /Ⅱ),moesin-like BCL2-induced protein1(Beclin1),phosphatase and tensin (PTEN) homologinduced putative kinase 1(PINK1) and dynamin-related protein Ⅰ (DRP1) was determined by Westem blotting.RESULTS More than 95％ of the cultured cells expressed vascular endothelial cadherin and herein were vascular endothelial cells.The MTT result showed that the cell survival of HUVECs was significantly decreased after incubation with CdTe QD (0.1-100 mg,L-1) for 24 h (P＜0.05,P＜0.01).CdTe QD also induced extensive fragmentation of the mitochondrial network.The results of JC-1 assay showed that CdTe QD (0.1-100 mg· L-1) caused the disruption of MMP.The percentage of HUVECs with higher MMP was reduced from (91.8±0.6)％ in cell control group to (90.2±1.1)％,(84.4±0.9)％ (P＜0.05) and (78.1 ±1.3)％ (P＜0.01),respectively.The Western blotting data suggested that CdTe QD 10 mg·L-1 significantly increased the expression of autophagy-related protein beclin 1 and the ratio of LC3 Ⅱ/LC3 Ⅰ (P＜0.05,P＜0.01),CdTe QD 1 mg· L-1 also raised the level of mitophagy-related proteins like PINK1 and DRP1.CONCULSION CdTe QD can induce mitochondrial dysfunction as well as mitophagy in HUVECs.

Objective To investigate the relationship between portal vein pressure and liver regeneration after 90% portal branch ligation in rats.Methods Forty-five male SD rats underwent 90% portal branch ligation (including 5 rats underwent sham operation),and then the changes of portal vein pressure and weight of unligated hepatic lobes were detected.The morphological changes of hepatocytes of the unligated hepatic lobes were observed under a light microscope.Proliferative cell nuclear antigen(PCNA)index was detected by immunohistochemistry,and hepatocyte apoptosis of the unligated hepatic lobes by TUNEL method.All data were analyzed by Pearson rank correlation analysis and t test.Results Thirty-eight out of 40 rats survived(95%).The ligated hepatic lobes diminished progressively,whereas the unligated hepatic lobes regenerated.Preoperative portal vein pressure was(9.1±1.8)cm H_2O(1 cm H_2O=0.098 kPa),and it was increased significantly shortly after the ligation and reached (15.8±2.7)cm H_2O 12 hours later(t=6.847,P＜0.05).The portal vein pressure decreased from(13.6±2.3)cm H_2O at day 1 to(9.3±2.0)cm H_2O at day 28.Preoperative positive PCNA index was 7%±3%,which was significantly lower than 14%±5%at postoperative 12 hours,21%±6%at day 3 and 26%±7%at day 5(t=9.129,P＜0.05),and it began to return to normal at day 5.Few apoptotic hepatoeytes were observed in preoperative liver tissue and unligated hepatic lobes.The expression of PCNA in unligated hepatic lobes and portal vein pressure had apositive correlation at postoperative day 1,3,5(r=0.913,0.896,0.908,P<0.05)and a negative correlation at postoperative day 14(r=-0.926,P<0.05).Conclusions The regeneration of hepatocytes in unligated hepatic lobes is activated after 90% portal branch ligation,and the regenerated liver compensates the weight loss of the ligated hepatic lobes.Liver is regenerated mainly by speeding hepatocyte proliferation rather than reducing hepatocyte apoptosis.Changes of portal vein pressure may play an important role in liver regeneration.

1.8.PCR was performed using the human Gpx-1 gene,and we obtained the needed target gene fragments.Conclusion The alkaline lysis extraction method is reliable for obtaining high quantities of DNA from blood clot suited for PCR amplification.

Objective To study the microinvasive treatment of post-gastrectomy acute cholecystisis. Methods Teweenty-eight cases of post-gastrectomy acute cholecystitis were treated by routine non-operative method first, if cases with no apparent relief after 24h of treatment were further treated with percutanous transhepatic gallbladder puncture and drainage(PTGD) guided by ultrasonography. Results Five cases were treated by non-operative method with complete relief within 24 hours. PTGD was done successfully in all the other 23 cases.In cases with ideal bile drainage at the time of intubation, the patients had immediate marked relief of their symptoms and signs, and they fully recovered within 3~7 days.Conclusions Ultrasound guided PTGD is an effective treatment for post-gastrectomy acute cholecystitis with the advantages of microinvasion and quick patient recovery.

To seek the optimum experiment methods of animal immunization with HCV gene and to explore the effect on antibody responses in mice immunized by pCD-HCV1 recombinant in different administration, recombinant pCD-HCV1 was constructed by technique of molecular biology and was injected into muscles of Balb/c of mice with different times, routes and dosage of inoculations as well as different treatment. The results showed that the serum antibody level reached 0.183±0.06,0.428±0.05,0.707±0.08 and 0.773±0.07(OD410 value) respectively after recombinant pCD-HCV1(100μg/mouse) were injected into mice once, twice, three times and four times. The antibody level of mice (n＝12) with four times inoculation was the highest; pCD-HCV1 was perfused into stomach orally in mice or were into mice by i.p, s.c and i.m(100μg/mouse, three times) in different routes (n＝6), and the antibody levels were 0.138±0.05, 0.178±0.07, 0.233±0.08 and 0.691±0.05 respectively; after the mice (n＝8) were inoculated with the pCD-HCV1 of different dosage(10μg, 50μg and 100μg) the antibody levels of three groups were 0.11±0.09, 0.33±0.04, and 0.700±0.07, and the results showed a significant difference (P＜0.01); Mice was injected with procaine (100μl, 0.4mg) by i.m or s.c. Then pCD-HCV1 was injected into mice and antibody levels were higher than that of mice immunized directly with recombinant pCD-HCV1 of same dosage. The results may provide a reference data deserved for screening the optimum immunization method of development HCV-DNA-based vaccine in mice model.