Objective Infantile spasms ( IS ) is the most common intractable epileptic encephalopathy during infancy,but there is the lack of animal experiment. By building IS animal model, the study discusses whether high-dose methylprednisolone has the protective effect on the immature rats of infantile spasms. Meth-ods Sprague-Dawley immature rats were randomly assigned to 3 groups on postnatal day 10 ( P10 ):control group、model groupⅠand model groupⅡ,thirty rats in each group. Immature rats in model groupⅠand mod-el group Ⅱ were injected NMDA intraperitoneally to induce seizures. The rats in the model groupⅡwere injec-ted intraperitoneally methylprednisolone on postnatal day 11,12 and 13. The clinical behavior of rats were ob-served and recorded. Neuronal apoptosis was detected by TUNEL. Immunohistochemistry and real-time PCR were used to analyze the expression of Bax,Bcl-2,caspase-3 in hippocampus. Results (1)83. 3% of rats in model groupⅠhad seizures,and none of rats in model groupⅡhad seizures on postnatal day 13. (2)The apop-totic cell number of brain tissues:model group Ⅱ( 14. 37 ± 2. 02 ) were lower than model group Ⅰ( 25. 67 ± 1. 52)and higher than control group(9. 00 ± 2. 50),the difference was statistically significant(all P<0. 05). (3)Bax protein and mRNA expression levels in model group Ⅱ(44. 55 ± 3. 58,2. 35 ± 1. 01)were lower than model group Ⅰ(58. 05 ± 4. 62,3. 27 ± 0. 95)and higher than control group (28. 90 ± 5. 14,1. 68 ± 0. 50),there was significant difference(all P<0. 05);(4)Caspase-3 expression levels of mRNA in model groupⅡ(5. 99 ± 1. 75) were lower than those in group model group Ⅰ(7. 88 ± 1. 60) and higher than those in control group (3. 60 ± 1. 70),there was significant difference(all P<0. 05). Conclusion High-dose methylprednisolone can reduce NMDA-induced seizures in the IS immature rats. High-dose methylprednisolone has protective effect on the NMDA-induced IS immature rats,which may be relation to weakening seizures and decreasing apoptosis.

Objective:To evaluate the efficacy of intravitreal ranibizumab (IVR) injection and/or laser photocoagulation on diabetic macular edema (DME) of different morphologic patterns based on optical coherence tomography (OCT).Methods:A non-randomized controlled clinical trial was conducted.A total of 79 diabetic patients (108 eyes) who were diagnosed as DME in Affiliated Hospital of Nanjing University of Chinese Medicine from March 2017 to February 2018 were enrolled.The subjects were divided into diffuse macular edema (DRT) group (41 eyes), cystoid macular edema (CME) group (37 eyes) and serous retinal detachment (SRD) group (30 eyes) according to the morphological characteristics of OCT, and received intravitreal injection of 0.05 ml (0.5 mg) ranibizumab and/or laser photocoagulation according to treatment guidelines.Best corrected visual acuity (BCVA), central macular thickness (CMT) of the subjects were recorded before treatment and 1 month, 3, 6 and 12 months after treatment.The morphologic changes of macular edema and complications were recorded.This study protocol adhered to the Declaration of Helsinki and was approved by an Ethics Committee of Affiliated Hospital of Nanjing University of Chinese Medicine (No.2017NL-13-03). Written informed consent was obtained from each patient before any medical examination and treatment.Results:The 1-, 3-, 6- and 12-month post-treatment average BCVA (LogMAR) of the DRT, CME and SRD groups were improved in comparison with before treatment, and the average CMT of the three groups at various time points after treatment was reduced than that before treatment (all at P<0.05). For the 39 eyes who received IVR treatment, the 12-month post-treatment average BCVA (LogMAR) of the DRT group was 0.41±0.40, which was significantly better than 0.60±0.40 of the CME group ( P=0.039). The 12-month post-treatment CMT of the DRT group was (286.05±109.56) μm, which was significantly thinner than (338.30±101.87)μm of the SRD group ( P=0.045). For the 69 eyes who received IVR combined with laser photocoagulation treatment, the 6- and 12-month post-treatment average BCVA (LogMAR) of the DRT group were significantly better than those of the CME group ( P=0.048, 0.043), and the average CMT at 12 months after treatment in the DRT group was (304.59±106.66)μm, which was significantly smaller than (369.34±107.80)μm in the SRD group, showing a statistical significance ( P=0.041). During the follow-up, 5 eyes with SRD turned to DRT, and 3 SRD eyes turned to CME.No eye changing from DRT and CME to SRD was found. Conclusions:Intravitreal ranibizumab injection and/or laser photocoagulation can significantly improve BCVA and reduce CMT of DME patients, and the efficacy is better in eyes with DRT than eyes with SRD or CME.

Objective:To explore the expression of microRNA-4695-5p in the serum of colorectal cancer patients and its effect on the proliferation and invasion of colorectal cancer CACO-2 cells.Methods:A total of 43 serum samples of colorectal cancer patients who were admitted to the Department of Gastrointestinal Surgery, Luoyang Central Hospital Affiliated to Zhengzhou University from March 2018 to November 2021 were selected, and serum samples of 43 healthy people who underwent outpatient physical examination were used as controls. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression levels in the serum of colorectal cancer patients and those of healthy individuals. The miR-4695-5p overexpression plasmid or the negative control plasmid were transfected into CACO-2 cells, namely the miR-4695-5p group and the control group, and the transfection efficiency was verified by qRT-PCR. CCK8 method and Transwell experiment were used to detect the effect of overexpression of miR-4695-5p on the proliferation and invasion of CACO-2 cells. The dual luciferase reporter gene experiment was used to verify the targeted binding relationship between miR-4695-5p and Ras-related C3 botulinum toxin substrate 1 (RAC1). qRT-PCR and Western blot were used to detect the effect of overexpression of miR-4695-5p on the expression of RAC1 gene and Wnt/β-Catenin signaling pathway protein.The software of SPSS28.0 was used to conduct data analysis. The measurement data of normal distribution were espressed by Mean±SD. The t-test was used to compare the means between two groups, and the one-way analysis of variance was used to compare the means of multiple groups. Results:The expression level of miR-4695-5p in the serum of patients with colorectal cancer was significantly lower than that of healthy individuals ( P<0.01). The relative expression levels of miR-4695-5p in the control group and miR-4695-5p group were 1.09 ± 0.65 and 8.83±2.03, respectively. The expression level of miR-4695-5p in CACO-2 cells in the miR-4695-5p group was 8.10 times that of the control group, and CACO-2 cells overexpressing miR-4695-5p were successfully constructed ( P<0.01). Compared with the control group, the proliferation ability of CACO-2 cells in the miR-4695-5p group was significantly reduced ( P<0.05), and the invasion ability of CACO-2 cells was significantly reduced ( P<0.01). Bioinformatics tools and dual luciferase reporter gene experiments confirmed that miR-4695-5p can target and bind RAC1 ( P<0.01). Compared with the control group, the expression of RAC1 gene in the miR-4695-5p group was significantly decreased ( P<0.01), and the expression of Wnt/β-Catenin signaling pathway proteins Wnt3a, β-catenin, and c-MYC decreased significantly. Conclusions:miR-4695-5p is lowly expressed in the serum of colorectal cancer patients. miR-4695-5p inhibits the proliferation and invasion of colorectal cancer CACO-2 cells by targeting the inhibition of RAC1 gene expression and down-regulating the activity of the Wnt/β-Catenin signaling pathway.

Tripterygium glycosides tablet(TGT),the classical commercial drug of Tripterygium wilfordii Hook.F.has been effectively used in the treatment of rheumatoid arthritis,nephrotic syndrome,leprosy,Behcet's syndrome,leprosy reaction and autoimmune hepatitis.However,due to its narrow and limited treatment window,TGT-induced organ toxicity(among which liver injury accounts for about 40％of clinical reports)has gained increasing attention.The present study aimed to clarify the cellular and molecular events underlying TGT-induced acute liver injury using single-cell RNA sequencing(scRNA-seq)technology.The TGT-induced acute liver injury mouse model was constructed through short-term TGT exposure and further verified by hematoxylin-eosin staining and liver function-related serum indicators,including alanine aminotransferase,aspartate aminotransferase,alkaline phosphatase and total bilirubin.Using the mouse model,we identified 15 specific subtypes of cells in the liver tissue,including endothelial cells,hepatocytes,cholangiocytes,and hepatic stellate cells.Further analysis indicated that TGT caused a significant inflammatory response in liver endothelial cells at different spatial locations;led to marked inflammatory response,apoptosis and fatty acid metabolism dysfunction in hepatocytes;activated he-patic stellate cells;brought about the activation,inflammation,and phagocytosis of liver capsular macrophages cells;resulted in immune dysfunction of liver lymphocytes;disturbed the intercellular crosstalk in liver microenvironment by regulating various signaling pathways.Thus,these findings elaborate the mechanism underlying TGT-induced acute liver injury,provide new insights into the safe and rational applications in the clinic,and complement the identification of new biomarkers and ther-apeutic targets for liver protection.

ObjectiveTo investigate the pharmacodynamic characteristics and explore the molecular mechanism of Honghua oral liquid （HOL） in relieving neuropathic pain （NP）. MethodHealthy male SD rats were randomly assigned into sham group， model group， low-， medium-， high-dose （0.5， 1.0， 2.0 mL·kg^-1·d^-1， respectively） HOL groups， and a positive drug （pregabalin， 25 mg·kg^-1·d^-1） group， with 6 rats in each group. Spinal nerve ligation （SNL） of L5 was conducted in other groups except the sham group. Drug administration was performed 3 days after the SNL surgery for 2 consecutive weeks， and samples were collected after the end of the administration. During the treatment period， the mechanical pain threshold and cold pain threshold were determined to measure the pain-relieving effect of HOL. Transcriptome sequencing was performed on hippocampal tissue samples from the sham， model， and high-dose HOL groups， and differentially expressed genes between the sham group and the model group as well as the model group and HOL high-dose group were obtained. After pathway enrichment analysis， we selected the targets which were closely related to neuroinflammation for validation， and predicted the specific binding sites of the major active components in HOL with the targets through molecular docking. In addition， the serum levels of tumor necrosis factor-α （TNF-α） and interleukin-10 （IL-10） were determined by enzyme-linked immunosorbent assay （ELISA） to evaluate the effect of HOL on neuroinflammation in NP rats. ResultCompared with the sham group， SNL decreased the mechanical pain threshold and cold pain threshold （P<0.05）. Compared with the model group， HOL recovered the mechanical pain threshold and cold pain threshold （P<0.05）. The transcriptome data showed that 376 differentially expressed genes （DEGs） were identified between the model group and the sham group， including 124 upregulated genes and 252 downregulated genes， and 194 DEGs between the model group and the high-dose HOL group， including 33 upregulated genes and 161 downregulated genes. Among them， insulin-like growth factor 1（IGF1）， matrix metallopeptidase-2 （MMP-2）， matrix metallopeptidase-14 （MMP-14）， erb-B2 receptor tyrosine kinase 2 （ERBB2）， and integrin subunit alpha 5 （ITGA5） associated with NP were selected for further validation. The Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） results showed that compared with the sham group， the modeling up-gurelated the mRNA levels of the above five molecules in the hippocampus （P<0.01）. Compared with model group， HOL down-regulated the mRNA levels of these molecules （P<0.01）. The molecular docking results showed that the main active components of safflower， hydroxysafflor yellow A， kaempferol， and quercetin， formed stable hydrogen bonds with the amino acid residues of IGF1， MMP-2， MMP-14， ERBB2， and ITGA5. The enzyme-linked immunosorbent assay（ELISA） results showed that compared with those in the sham group， the serum levels of TNF-α and IL-10 were out of balance in the model rats （P<0.01）. Compared with the model group， HOL lowered the level of the pro-inflammatory cytokine TNF-α （P<0.01） and elevated that of the anti-inflammatory cytokine IL-10 （P<0.05）. ConclusionHOL exerts analgesic effect on SNL rats by inhibiting neuroinflammation.