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.

Objective To explore the role of Adra1a in regulating the LPS-induced inflammation response in primary hepatocytes of lipopolysaccharide-binding protein knockout(Lbp-/-)mice.Methods Primary hepatocytes were extracted from WT and Lbp-/-mice using a two-step perfusion method,and an inflammation model was established using LPS induction.Expression of Adra1a in primary hepatocytes of Lbp-/-mice was suppressed by administering the inhibitor prazosin and transfection with si-Adra1a.The cells were divided into three groups under inhibitor conditions:control group A,LPS group A,and prazosin group.For siRNA transfection,cells were also divided into groups:control group B,LPS group B,si-NC group,and si-Adra1a group.WT primary hepatocytes were divided into two groups:control group(blank)and LPS group(12 h stimulation).Changes in the Adra1a response to LPS stimulation were verified by Western blot.Other method ologies,such as CCK-8,qRT-PCR,and Western blot assays,were used to confirm improvements in cell inflammation and the survival rate by prazosin and si-Adra1a.Results Significant elevation in Adra1a protein expression in Lbp-/-primary hepatocytes was observed post-LPS stimulation(P＜0.01),whereas no notable change was found in the wildtype.A remarkable increase in the cell survival rate was noted in prazosin and si-Adra1a groups(P＜0.01,P＜0.05).Furthermore,prazosin and si-Adra1a groups exhibited significantly reduced expression of proinflammatory factors TNF-αand IL-1 β(P＜O.01),p-p38,p-ERK,and p-JNK(P＜0.01),which are associated with cell damage and inflammation.Conclusions Following LPS stimulation,upregulation of Adra1a and proinflammatory cytokine expression was observed in Lbp-/-primary hepatocytes.Specific downregulation of Adra1a expression using prazosin and si-Adra1a significantly decreased LPS-induced proinflammatory cytokines in Lbp-/-primary hepatocytes.Adra1a is implicated in the regulation of the LPS-induced inflammation response in primary hepatocytes of Lbp-/-mice.

Objective:To systematically review the central line associated bloodstream infection prediction models based on machine learning.Methods:The research on central line associated bloodstream infection prediction models based on machine learning was systematically searched in Cochrane Library, PubMed, Embase, WanFang Data, VIP, and China National Knowledge Infrastructure. The search period was from database establishment to March 18, 2022. Two researchers independently screened articles and extracted data. The data extraction was carried out using CHARMS checklist. The prediction model risk of bias assessment tool (PROBAST) was used to evaluate the bias risk of included studies.Results:A total of 9 related studies were ultimately included. The overall results indicated that modeling using machine learning algorithms had higher predictive accuracy compared to traditional Logistic regression. There was a risk of multiple biases in the modeling process, but the overall applicability evaluation of the article performed well.Conclusions:Existing evidence suggests that machine learning can accurately predict the occurrence of central line associated bloodstream infections, but there is a certain bias in development and reporting. In the future, emphasis should be placed on external validation and model updates, continuously improving its predictive performance.

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 investigate the efficacy of balloon dilatation in the treatment of traumatic biliary stricture(TBS).Methods Eighty-one patients with TBS who had been admitted into General Hospital of Chengdu Command from February 1997 to February 2007 were divided into 2 groups.In the control group,40 patients underwent choledoco-jeiunostomy with T tube stenting in the anastomotic stoma for 6 months.In the balloon dilatation group,41 patients received implantation of the balloon catheters along with the two arms of the T tube in the anastomotic stoma.The efficacy of different surgical procedures were analyzed by chi-square test.Results Seven patients in the control group were failed with a failure rate of 18%.Of the 7 palients,6 were transferred to receive balloon dilatation and 5 were cured.In the balloon dilatation group,only 1 patient was failed and the failure rate was 2%.The diameter of the anastomotic stoma ilq the balloon dilatation group was dilated gradually to 12 mm in 3 months.The difference upon the success rate of the operation between the 2 groups had statistical significance (x2=5.10,P＜0.05).Conclusions Balloon dilatation is effective in the treatment of TBS.

Objective To evaluate the effect s of Ginkgo Leaf Extract and diphyridamole injection on patient s with CKD.Methods 60 patients with CKD were made up with 36 cases in Ⅲ phase and 24 cases in Ⅳ phase.The patients with CKD were given Ginkgo Leaf Extract and diphyridamole injection,20 mL/d,for 15 days.The parameters of clini- cal condition,function of kidney,ALB and blood fat were observed before and after the therapy and the data were ana- lyzed.Results There were 28 cases that clinical condition was better; 24cases no difference; and 8 cases worse.The level of TC and TG were significantly decreased after the therapy,as well as the uric protein quality of 24 hours(P0.05). Conclusion Essential treatment for CKD combined with Ginkgo Leaf Extract and diphyridamole injection can improve nutrition,delay the process of CKD and protect the renal function in patient s with Ⅲ phase.

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.

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.