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Objective To investigate the influence of transient receptor potential cation channel 6 (TRPC6) on renal tubulointerstitial inflammation in mice with diabetic kidney disease (DKD) and its mechanism. Methods A total of 36 male C57/BL6J mice, aged 6 weeks, were randomly divided into control group (group A), DKD model group (group B), DKD+normal saline intervention group (group C), DKD+mitophagy activator intervention group (group D), DKD+negative control lentivirus transfection group (group E), and DKD+TRPC6 knockdown lentivirus transfection group (group F), with 6 mice in each group. The mice in groups A and B were respectively given intraperitoneal injection of 0.1 mmol/L citrate buffer and 10 g/L streptozotocin (hereinafter referred to as administration); the mice in groups C and D were respectively given normal saline and 10 mmol/L urolithin A by gavage in addition to the treatment in group B; the mice in groups E and F were respectively injected with negative control lentivirus and TRPC6 knockdown lentivirus via the tail vein in addition to the treatment in group B. The levels of fasting blood glucose (FBG), urinary albumin-to-creatinine ratio (ACR), and blood urea nitrogen (BUN) were measured at week 12 after administration; PAS staining was used for the observation and scoring of renal tubular injury; RT-qPCR was used to measure the mRNA expression levels of inflammatory factors [interleukin-1β (IL-1β), monocyte chemoattractant protein-1 (MCP-1), and tumor necrosis factor-α (TNF-α)] in renal tissue; immunohistochemical staining was used to measure the protein expression level of TRPC6 in renal tissue, and Western blotting was used to measure the relative expression levels of TRPC6, LC3B, P62, PINK1, and Parkin in renal tissue; transmission electron microscopy was used to observe the change in the number of mitophagosomes in renal tubular cells of mice. HK-2 cells were divided into high glucose+TRPC6 siRNA transfection+DMSO intervention group (group G, treated with TRPC6 siRNA+35.0 mmol/L glucose+0.06% DMSO) and high glucose+TRPC6 siRNA transfection+mitophagy inhibitor intervention group (group H, treated with TRPC6 siRNA+35.0 mmol/L glucose+12 μmol/L oroxylin A), and then RT-qPCR was used to measure the mRNA expression levels of inflammatory factors (IL-1β, MCP-1, and TNF-α) in cells. Results At week 12 after administration, compared with group A, group B had significantly higher whole blood FBG, ACR, BUN, renal tubular injury score, mRNA expression levels of inflammatory factors, and protein expression levels of TRPC6 and P62 in renal tissue (t=2.77-13.61,P

Objective To investigate the method for establishing an immortalized mouse corneal epithelial cell line, and to identify the stability of its phenotype. Methods Primary mouse corneal epithelial cells were extracted and transfected with Simian Vacuolating Virus 40 Large T Antigen (SV40-LT) lentivirus to establish an immortalized cell line. HE staining and immunofluorescent staining were used to identify the phenotypes and markers (K12, Pax6, Ki-67) of corneal epithelium. Primary cor-neal epithelial cells were extracted to identify K14 protein in corneal epithelial cell layer, and bright field observation was performed to evaluate cell morphological characteristics. Several markers (K12, Pax6, Ki-67, ABCG2, K14) were identified for P1 and P8 cells after transfection. Crystal violet staining was used to observe the colony-formation ability of P1 and P8 cells after transfection. Results Mouse corneal epithelium was stained positive for K12, Pax6 and Ki-67, while the P1 and P8 generations of mouse cor-neal epithelial cells were stained positive for K12, Pax6, Ki-67, K14, and ABCG2; there were no significant differences in the positive rates of the above markers (P>0.05). There was no significant difference in colony-formation ability between P1 and P8 cells (P>0.05). Conclusion The immortalized mouse corneal epithelial cell line is successfully established by transfection with SV40-LT lentivirus, which has stable phenotype and proliferation.

Objective To investigate the neuroprotective effects and mechanism of CMS121 in a neuron model of oxygen-glucose deprivation/reperfusion (OGD/R) and a mouse model of ischemia/reperfusion (I/R). Methods Cortical neurons were divided into control group (normal culture), OGD/R group (OGD/R model construction), and OGD/R+CMS121 group (OGD/R model construction plus treatment with CMS121). All neurons’ viability and lactate dehydrogenase (LDH) release were assessed after 24 h of reperfusion. Western blotting was performed to measure the expression of protein kinase B (Akt) and phosphorylated Akt (p-Akt) in the neurons after 6 and 24 h of reperfusion. C57BL/6J mice were randomly divided into sham group (sham operation), I/R group (I/R model construction), and I/R+CMS121 group (I/R model construction plus treatment with CMS121). After 24 h of reperfusion, we assessed the neurological damage of the mice with the Neurological Severity Score (NSS), and calculated the brain infarct volume (as a percent of total brain volume) after 2,3,5-triphenyltetrazolium chloride staining. Results There were significant differences in cell viability and LDH release between the three neuron groups (F=71.21,88.93,P0.05). After 6 h reperfusion, a significant difference was observed in neuronal p-Akt expression between the three neuron groups (F=18.78,P0.05). After 24 h reperfusion, p-Akt expression differed significantly between the three neuron groups (F=38.50,P0.05). There were significant differences in the NSS score and infarct volume between the three mouse groups (F=20.24,118.60,P

ABSTRACT Objective To evaluate the morphology and morphometry of the muscles extensor digitorium longus and soleus of C57BL/6 females, who were exposed to glyphosate during pregnancy and lactation. Methods Twelve female mice from the C57BL/6 lineage were used. After detection of pregnancy, they were divided into a Control Group, which received only water, and a Glyphosate Group, which received water with 0.5% glyphosate during pregnancy and lactation. Both groups received ad libitum standard diet. After weaning, the females were euthanized and weighed; naso-anal length was measured, and fats were collected and weighed. The muscles extensor digitorium longus and soleus were collected, and their length and weight were measured. Then, the muscles were fixed in Methacarn to perform the histological study of muscle fibers. Results Glyphosate Group presented lower weight gain during pregnancy and also lower final body weight and naso-anal length; however, the other body parameters evaluated did not present a significant difference in relation to the Control Group. Significant differences were also not observed in the analysis of muscle fibers and connective tissue. Conclusion Exposure to 0.5% glyphosate during pregnancy and lactation resulted in lower weight gain during pregnancy, final weight, and naso-anal length. Despite not directly altering the morphology of muscle tissue, these results may indicate enough exposure to interfere with animal metabolism.

Different lines of investigation suggest that the medial amygdala is causally involved in the processing of information linked to social behaviour in rodents. Here we investigated the consequences of temporary inhibition of the medial amygdala by bilateral injections of lidocaine on long-term social recognition memory as tested in the social discrimination task. Lidocaine or control NaCl solution was infused immediately before learning or before retrieval. Our data show that lidocaine infusion immediately before learning did not affect long-term memory retrieval. However, intra-amygdalar lidocaine infusions immediately before choice interfered with correct memory retrieval. Analysis of the aggressive behaviour measured simultaneously during all sessions in the social recognition memory task support the impression that the lidocaine dosage used here was effective as it – at least partially – reduced the aggressive behaviour shown by the experimental subjects towards the juveniles. Surprisingly, also infusions of NaCl solution blocked recognition memory at both injection time points. The results are interpreted in the context of the importance of the medial amygdala for the processing of non-volatile odours as a major contributor to the olfactory signature for social recognition memory.

La corriente eléctrica directa (CED) ha sido utilizada en el tratamiento de tumores sólidos. Se realizó este estudio para demostrar el efecto antitumoral de ésta en un fibrosarcoma Sa-37 mediante hallazgos tumorales y peritumorales. Se le aplicaron 5 mA durante 30 min a un grupo de tumores, a través de electrodos de platino insertados en ellos. A otro grupo no se les suministró CED. Se observó una respuesta inflamatoria aguda y un retardo en los crecimientos en los tumores tratados con respecto a los no tratados. Se concluyó que la CED tiene una acción retardadora en el crecimiento de los tumores tratados por los altos porcentajes de necrosis inducidos en éstos y la estimulación del sistema inmune.Direct electric current (DEC) has been used in the treatment of the solid tumors. The aim of this study is demonstrate the antitumoral effect of it on a fibrosarcome Sa-37 through tumor and peritumoral findings. 5 mA during 30 minutes to a tumors group was applied through platinum electrodes inserted into it. Another group of tumors it not was applied DEC. It was observed in the treated tumors with respect to the untreated an acute inflammatory response and a growth delay of it. It was concluded that DEC has a delay action in the treated tumors growth due to the necrosis percentage high induced into the tumor and the stimulation of the immune system.