Objective To study the effect of paeoniflorin on inflammatory chemokines and their receptor in a rat model of ovalbumin-induced asthma.Methods Sixty 6-week old SPF female BALB/c mice were used in this study.To es-tablish a mouse model of asthma by sensitizing and challenging with ovalbumin.ELISA was used to analyze the serum IL-6 and TNF-α, and the specific IgE against ovalbumin ( OVA-IgE ) , CCL19 and CCL21 in bronchoalveolar lavage fluid (BALF).RT-PCR was performed to determine CCR7mRNA and protein expression of chemokine receptor CCR7, and the level of NF-κB was tested by Western blot.ResultsIn In the paeoniflorin groups, the serum IL-6 and TNF-αlevels were significantly lower, and the OVA-IgE, CCL19 and CCL21 levels in BALF were significantly reduced, compared with that in the control group.CCR7mRNA and protein expression of chemokine receptor CCR7 and NF-κB in the lung were significant-ly reduced by paeoniflorin.Conclusions Paeoniflorin has a remarkably inhibitory effect on the airway inflammatory chemo-kines CCL19/CCL21 and the receptor CCR7 in the mouse model of asthma.

Objective Ischemia reperfusion injury (IRI) is a major limiting factor of graft survival in organ transplantation.We've established a novel procedure called ischemia-free liver transplantation (IFLT) in big animal study.In this report,we aimed to investigate the feasibility and early outcomes of IFLT.Methods We have performed 3 cases of IFLT during July 23,2017 to August 9,2017.We analyzed the surgical methods,normothermic perfusion parameters,blood gas analysis,liver function tests and complications early after liver transplantation.Pathologic studies and immunohistochemical staining of donor liver biopsies were conducted before procurement,at the end of machine perfusion,as well as after re-vascularization for evaluating IRI.Results The surgical procedures of all 3 patients were successful,without stoppage of blood supply for the liver grafts throughout organ procurement,ex vivo preservation and implantation.During normothermic perfusion,the pH value was stable within the normal range and the lactate levels dropped quickly to lower than detected (＜0.3 mmol/L) within 1.5-3 h.The livers continued to produce bile with the volume of 2-6 mL/h.Hematoxylin and eosin (HE) staining evaluation and TdT-mediated dUTP nick end labeling (TUNEL) assay of biopsies taken from liver tissues before procurement,at the end of machine perfusion and after re-vascularization,showed few necrostic and apoptotic hepatocytes in the liver biopsies.The immunohistochemical staining of IL-1β and vWF suggested no inflammatory cytokine release and sinusoidal endothelial cell activation.The three patients recovered smoothly without rejection,vascular and biliary complications.Conclusion IFLT is a feasible and effective procedure,which is able to overcome the major limitations of conventional procedure.The novel IFLT will become one of the mainstream transplant procedures in the future.

Objective To investigate the correlation between the elastographic characteristics of liver and postoperative function of liver allografts.Methods Forty-eight cases of liver transplantation from The First Affiliated Hospital of Sun Yat-sen University were analyzed,Shear wave elastography (SWE) was performed before operation or at one week or one month post-operation.Liver function was evaluated by measuring alanine aminotransferase (ALT),aspartate transaminase (AST),total bilirubin (TBIL),γ-glutamine transferase (GGT),albumin (ALB),alkaline phosphatase (ALP),prothrombin time (PT),activated partial thromboplastin time (APTT),and international normalized ratio (INR).Early allograft dysfunction (EAD) was also analyzed with reference to SWE among liver transplant recipients.Results SWE at one week after transplantation was significantly correlated with TBIL (r=0.525 6,P<0.01),APTT (r=0.668 3,P<0.000 1),PT (r=0.593 7,P=0.000 1),INR (r=0.609 6,P<0.000 1) and prealbumin (r=-0.464 1,P<0.01).However,no significant correlation was observed between pre-operative SWE and parameters of post-operative liver function.SWE in EAD patients was higher than that of patients without EAD (17.60±1.09 kPa vs.13.38±0.99 kPa,P<0.01).The optimal cut-off value of SWE at one week post-operation was 14.85 kPa.Conclusion Postoperative SWE is significantly correlated with postoperative liver function tests and EAD,suggesting SWE is a potential test for evaluating the quality of liver allografts.

BACKGROUND:Exercise training can improve osteoporosis,but its effects and mechanisms on senile osteoporosis are not fully understood. OBJECTIVE:To observe the effect of treadmill exercise on osteoporosis and wnt/β-catenin signal pathway in aged rats. METHODS:Sixteen 24-month-old male Sprague-Dawley rats were randomly divided into osteoporosis group(n=8)and treadmill group(n=8)and eight 6-month-old male Sprague-Dawley rats were used as young control group.The model of senile osteoporosis was replicated by natural aging and the rats in the treadmill group were treated with treadmill exercise once a day,5 days a week,for 8 weeks.Levels of bone metabolic markers such as type I collagen cross-linked C-terminal peptide,tartrate resistant acid phosphatase,osteocalcin and bone specific alkaline phosphatase were detected by ELISA;bone mineral density of the left femur and L5 was measured by dual energy X-ray;bone scanning and bone microstructure quantitative analysis were performed by bone micro-CT;and the mRNA and protein expression levels of wnt3a,β-catenin,LRP5,DKK1 and GSK3β were detected by RT-PCR and western blot,respectively. RESULTS AND CONCLUSION:Compared with the young control group,the osteoporosis group showed a reduction in serum bone specific alkaline phosphatase and osteocalcin levels(P<0.05),bone mineral density of the femur and L5,the number of tibia and L4 bone trabeculae,bone volume,bone volume fraction(P<0.05),and mRNA and protein expression of wnt3a,β-catenin,and LRP5 in bone marrow tissue(P<0.05)as well as an increase in serum levels of tartrate resistant acid phosphatase and type I collagen cross-linked C-terminal peptide(P<0.05),the intertrabecular space between the tibia and L4,structural model index(P<0.05),and mRNA and protein expression of DKK1 and GSK3 β in bone marrow tissue(P<0.05).In addition to the reduced number of trabeculae in the tibia and L4 vertebrae,the trabeculae were structurally disturbed and sparsely aligned and fractured.Compared with the osteoporosis group,the treadmill group showed an increase in serum bone specific alkaline phosphatase and osteocalcin levels(P<0.05),bone mineral density of the femur and L5(P<0.05),the number of tibial trabeculae,bone volume,bone volume fraction(P<0.05),mRNA and protein expression of wnt3a,β-catenin,and LRP5 in bone marrow tissue(P<0.05)but a reduction in the serum levels of tartrate resistant acid phosphatase and type I collagen cross-linked C-terminal peptide,L4 trabecular space,tibial trabecular space,structural model index,and mRNA and protein expression of DKK1 and GSK3 β in bone marrow tissue(P<0.05).In addition to the increased number of tibial and L4 trabeculae,the trabeculae were arranged in a regular and dense pattern and were connected to a network.To conclude,treadmill exercise may improve osteoporosis in aged rats by activating the wnt/β-catenin signal pathway.

Background Acute cadmium (Cd) exposure can cause damage to multiple tissues, with the kidney being the primary target organ. The development of Cd-induced acute kidney injury involves complex mechanisms, in which autophagy and oxidative stress play crucial roles. Objective To investigate the effect of 10-hydroxy-2-decenoic acid (10-HDA) on kidney injury in mice exposed to cadmium, and provide experimental basis for studying the pathogenesis and prevention of Cd poisoning. Methods Thirty-five male C57BL/6 mice were divided into 7 groups (each of 5 mice): control group (normal saline, intraperitoneal injection), CdCl₂ group (4 mg·kg⁻¹, intraperitoneal injection), intervention groups ( 4 mg·kg⁻¹ CdCl₂, intraperitoneal Injection + 50, 100, 150, or 200 mg·kg⁻¹ 10-HDA, oral gavage), and 10-HDA group (150 mg·kg⁻¹, oral gavage). All treatments were given for 14 d. Twenty-four hours after the last infection, physiological indicators [blood urea nitrogen (BUN), creatinine (CRE), malondialdehyde (MDA), and superoxide dismutase (SOD)], histopathological indicators, autophagy-related proteins (Atg7, Atg5, Beclin-1, and LC3), and mitochondrial autophagy-related proteins (PINK1 and Parkin) were detected to examine the effect of 10-HDA on kidney injury caused by CdCl₂. Results Compared with the control group, the body weight of mice in the CdCl₂ group was significantly reduced (P<0.01); compared with the CdCl₂ group, the body weight of mice after intervention with different concentrations of 10-HDA was significantly increased (P<0.01). CdCl₂ significantly increased BUN and CRE in the serum samples compared with the control group (P<0.01), which was significantly reduced to varying degrees after 100, 150, and 200 mg·kg⁻¹ 10-HDA intervention (P<0.01). MDA significantly increased and SOD significantly decreased in the renal cortex following CdCl₂ administration compared with the control group (P<0.01), which was resolved following 10-HDA administration at different concentrations (P<0.01). In histopathological studies, 10-HDA restored injured kidney tissues induced by CdCl₂. The expression levels of autophagy proteins Atg7 and LC3-II/I were significantly increased (P<0.05), and the expression level of Beclin-1 was significantly decreased (P<0.05) in the CdCl₂ group compared with the control group. The expression levels of Atg7 were reduced to varying degrees after treatment with designed concentrations of 10-HDA, the expression levels of LC3-II/I were also reduced in the 50, 150, and 200 mg·kg⁻¹ 10-HDA intervention groups, and the expression levels of Beclin-1 were increased in the 50, 100, and 150 mg·kg⁻¹ 10-HDA intervention groups (P<0.05). The expression levels of PINK1 and Parkin in the CdCl₂ group and the 50 mg·kg⁻¹ 10-HDA intervention group were lower than those in the control group (P<0.01). Compared with the CdCl₂ group, the expression levels of PINK1 increased to varying degrees after 100, 150, and 200 mg·kg⁻¹ 10-HDA intervention, and the expression levels of Parkin increased in all 10-HDA intervention groups (P<0.01). Conclusion The intervention using 10-HDA can lessen acute kidney injury caused by CdCl₂, reduce the expression of autophagy-related proteins, and increase the expression of mitochondrial autophagy-related proteins.

Brain metastasis is a common and serious complication of breast cancer, which is commonly associated with poor survival and prognosis. In particular, the treatment of brain metastasis from triple-negative breast cancer (BM-TNBC) has to face the distinct therapeutic challenges from tumor heterogeneity, circulating tumor cells (CTCs), blood-brain barrier (BBB) and blood-tumor barrier (BTB), which is in unmet clinical needs. Herein, combining with the advantages of synthetic and natural targeting moieties, we develop a "Y-shaped" peptide pVAP-decorated platelet-hybrid liposome drug delivery system to address the all-stage targeted drug delivery for the whole progression of BM-TNBC. Inherited from the activated platelet, the hybrid liposomes still retain the native affinity toward CTCs. Further, the peptide-mediated targeting to breast cancer cells and transport across BBB/BTB are demonstrated in vitro and in vivo. The resultant delivery platform significantly improves the drug accumulation both in orthotopic breast tumors and brain metastatic lesions, and eventually exhibits an outperformance in the inhibition of BM-TNBC compared with the free drug. Overall, this work provides a promising prospect for the comprehensive treatment of BM-TNBC, which could be generalized to other cell types or used in imaging platforms in the future.

Objective To produce rabbit polyclonal antibody against mouse testis expressed 38 (TEX38). Methods Full-length open reading frame sequence of TEX38 was amplified and inserted into the pET-30a-(+) vector to construct pET-30a-TEX38 prokaryotic plasmid. The recombinant plasmid was transformed into E.coli BL21, and expression was induced with isopropyl β-D-thiogalactopyranoside (IPTG). New Zealand white rabbits were immunized with TEX38 protein after purification and denaturation, then TEX38 polyclonal antibodies were collected from rabbit serum samples. ELISA was performed to detect the antibody titer. Western blot and immunofluorescence staining were performed to determine the specificity of TEX38 polyclonal antibodies. Results The pET-30a-TEX38 recombinant plasmid was constructed, and TEX38 prokaryotic protein was expressed and purified successfully. After immunization, the titer of TEX38 antibody reached 1:1 000 000. Western blot analysis and immunofluorescence staining showed that TEX38 was localized in the mouse spermatogenic cells and sperms with a good specificity. Conclusion The rabbit polyclonal antibody against mouse TEX38 is successfully produced, and the expression of TEX38 in mouse spermatogenic cells and sperms is validated.
Male ; Rabbits ; Animals ; Mice ; Testis ; Antibodies ; Enzyme-Linked Immunosorbent Assay ; Immunization ; Spermatozoa ; Escherichia coli

AL3810, a molecular dual inhibitor of the vascular endothelial growth factor receptor (VEGFR) and fibroblast growth factor receptor (FGFR), has earned the permission of phase II clinical trial for tumor treatment by China FDA. As a reversible ATP-competitive inhibitor, AL3810 targets ATP-binding site on intracellular region of VEGFR and FGFR, whereas, AL3810 lacking interplay with extracellular region of receptors rendered deficient blood-brain tumor barrier (BBTB) recognition, poor brain penetration and unsatisfactory anti-glioma efficacy. Integrin