Objective To explore whether extracellular signal-reg-ulated kinase (ERK) signaling pathway is involved in the hypoxia-inducible factor-1α (HIF-1α) expression in the hippocampus of epileptic rats.Methods A total of 208 21-d old SD rats were equally randomized into status epilepticusgroup (SE,n=96),normal control group (NS,n=96) and PD98059 (the ERKsignaling pathway specific inhibitor) treatment group (n=16),respectively; SE rat models of the SE group were induced by intraperitoneal injection of 1％ PTZ (80 mg/kg),and rats of the NS group received injection of normal saline (NS); 0.5,1,1.5,6,12 and 24 h after the inducement,the mRNA and protein expressions of HIF-lα and ERK1/2 in the hippocampus of rats in these two groups were examined by RT-PCR and Westem blotting.For rats in the PD98059 treatment group,PD98059 was intraperitoneally injected 10 minutes before intraperitoneal injection of pentylenetetrazol; 1 h after the inducement,the mRNA expressions of HIF-1α and ERK1/2 in the hippocampus of rats were examined by RT-PCR,while 1.5 h after the inducement,the protein expressions of HIF-1α and ERK1/2 in the hippocampus of rats were examined by Western blotting.The results of the PD98059 treatment group would be compared with those of the SE group.Results Compared with the NS group,the mRNA and protein expressions of HIF-1α and ERK 1/2 in the hippocampus of rats in the SE group after SE increased significantly; the peakexpression time ofERK1/2 mRNA was 1 h after SE (1.112±0.126 h),and the ERK1/2 protein mostly expressed at 1.5 h after SE (1.127±0.155 h).As to HIF-1α mRNA and its protein,the peak expression time was 1.5 h after SE (0.589±0.090 h) and 6 h after SE (0.230±0.052 h),respectively (P＜0.05).Compared with the SE group,the mRNA and protein expressions of HIF-1α and ERK1/2 in the hippocampus of all the rats in the PD98059 treatment group after SE decreased significantly (P＜0.05);positive correlation between HIF-1α and ERK1/2 mRNA in the SE group was noted (r=0.688,P=0.000).Conclusion ERK signaling pathway is activated in the epileptic rats and it participates in the expressionof HIF-1α in the hippocampus.

Objective　To compare the irradiation-protective and inter-synergestic effects of E838,WR-2721, Rubia cordifolia, cystamin e hydrochloride and ethinyl estradiol on radiation and combined radiation-burn injury. Methods　Above-mentioned drugs were given to the mice i ntraperitoneally, or intragastrcally, then, the mortality and the average surviv al d for 30 d were observed before and after the administration of the drug s. Results　①When drugs were before injury , the survival rate and the average survival d of the radiation and combined radiation-burn injured mice were increased obviously with the best effect in E838 and WR-2721. ②When drugs were given after injury, E838 and R. cordifolia also kept the effect. ③Combined appling WR-2721（pre) and E838（post）displayed a significant syner gistic reaction. Conclusion　E838 and WR-2721 are more e ffective than the others in the prevention of radiation.

OBJECTIVETo observe the change of apelin and its receptor (APJ) in the lung tissue of rats with pulmonary hypertension induced by monocrotaline and to explore its significance.

METHODSTwenty-five male SD rats were randomly divided into control group (n = 10) and monocrotaline group (n = 15). On the twenty-first day after the rats were intraperitoneally injected 60 mg/kg monocrotaline for monocrotaline group or equal volume vehicle for control group, the mean pulmonary artery pressure was measured by right heart catheterization. Histopathological study of lung tissue was done with hematoxylin-eosin (HE) and Masson's trichrome staining. The concentration of apelin in the plasma was measured by radioimmunoassay. The expressions of apelin/APJ proteins and genes in lung tissue were measured respectively by Western blot and reverse transcription polymerase chain reaction (RT-PCR).

RESULTSThe mean pulmonary arterial pressure, right ventricular hypertrophy, pulmonary vascular remodeling index, content of apelin protein in lung tissue of monocrotaline group were higher than those in control group. APJ protein and gene expression in monocrotaline group were significantly lower than those in control group (P < 0.01, P < 0.05), but apelin gene expression in the lung tissue between the two groups had no significant difference.

CONCLUSIONEndogenous apelin/APJ dysfunction may play an important role in the development of pulmonary hypertension induced by monocrotaline.

Animals ; Apelin ; Apelin Receptors ; Hypertension, Pulmonary ; chemically induced ; metabolism ; Intercellular Signaling Peptides and Proteins ; metabolism ; Lung ; metabolism ; Male ; Monocrotaline ; adverse effects ; Rats ; Rats, Sprague-Dawley ; Receptors, G-Protein-Coupled ; metabolism

Objective The aim of this research is to identify and compare the adjustment levels in patients with temporary and permanent colostomy, and to provide evidence to improve nursing performance. Methods In the First, Second, and Third Affiliated Hospital of Kunming Medical University, 52 patients with colorectal stoma participated in the research and completed the questionnaire of OAS (Ostomy Adjustment Scale) and demographic data. The data were established using Epidata 3.1, then analyzed with SPSS 17.0. The results were described using frequency, mean, stand deviations, and p values.Results Patients with permanent colostomy gained higher scores in OAS scores and psychological adaptation. Patients with temporary colostomy had higher scores in physical adaptation and social adaptation. Conclusion The nursing staff will promote the quality of life for this population from multiple perspectives in the future.

Objective:To explore the cellular heterogeneity and the differences in branched trajectory of pericytes between keloids and localized scleroderma, and to provide new clues for the pathogenesis and therapeutic targets of the two skin fibrotic diseases.Methods:Single cell transcriptome sequencing (scRNA-seq) data of 3 cases of scleroderma, 4 cases of keloid and their corresponding 4 cases of adjacent normal skin samples were selected from GEO and GSA-Human databases, and the expression matrix of the data was drawn. Seurat 4.3.0 of R (4.2.2) was used to process the t-distributed stochastic neighbor embedding ( t-SNE) visualization map. Monocle 2.24.0 was used to analyze the pseudo-temporal trajectory of pericytes. Results:The unsupervised clustering of keloid and scleroderma skin tissues revealed 19 different cell populations, among which C7 and C11 cells were pericytes, marked by high expression levels of PDGFRB and RGS5 genes, accounting for 7.53% of the total cells. Pericytes can be further divided into 8 subgroups. Pseudo-temporal analysis revealed a branched trajectory with two major branches, that is, cell fate 1 and cell fate 2, which could be further divided into 5 cellular states of pericytes (S1-S5). S4 constituted the most of the prebranch, which represented the cellular state of the initial pericyte phenotype. S5 constituted the most of the cell fate 1 branch, which represented the early differentiation state of the pericyte phenotype. S1, S2, S3 constituted the most of the cell fate 2 branch. S3 represented the intermediate differentiation state of the pericyte phenotype, while S1 and S2 represented the terminal differentiation states of the pericyte phenotype. Compared with the uniform distribution of various differentiation states of pericytes in normal skin, the keloid pericytes mainly distributed in the prebranch (S4), cell fate 1 (S5) and the first half of cell fate 2 (S3), representing cellular states of the initial, early and intermediate phases of the pericyte phenotype. Branched expression analysis modeling revealed the overexpression of SOX4, COL4A1, COL6A3, AHR, CXCL3 and IL1R1 genes, et cetera. On the other hand, the localized scleroderma pericytes mainly distributed in the bottom half of cell fate 2 (S1, S2), representing the final differentiated phase of pericyte phenotype, which overexpressed ACTA2 and MYH11 genes.Conclusion:Pericytes in keloid and scleroderma are heterogenous and have different differentiation trajectories. Pericytes in keloid have stem-like characteristics, and play an important role in the pathologic characteristics of invasiveness and recurrence through high expression of genes related to cell stemness, epithelial-mesenchymal transition, invasiveness, and immune microenvironment regulation. However, pericytes in localized scleroderma may mainly transdifferentiate into myofibroblasts, leading to their fibrotic pathological phenotype.

Objective:To explore the cellular heterogeneity and the differences in branched trajectory of pericytes between keloids and localized scleroderma, and to provide new clues for the pathogenesis and therapeutic targets of the two skin fibrotic diseases.Methods:Single cell transcriptome sequencing (scRNA-seq) data of 3 cases of scleroderma, 4 cases of keloid and their corresponding 4 cases of adjacent normal skin samples were selected from GEO and GSA-Human databases, and the expression matrix of the data was drawn. Seurat 4.3.0 of R (4.2.2) was used to process the t-distributed stochastic neighbor embedding ( t-SNE) visualization map. Monocle 2.24.0 was used to analyze the pseudo-temporal trajectory of pericytes. Results:The unsupervised clustering of keloid and scleroderma skin tissues revealed 19 different cell populations, among which C7 and C11 cells were pericytes, marked by high expression levels of PDGFRB and RGS5 genes, accounting for 7.53% of the total cells. Pericytes can be further divided into 8 subgroups. Pseudo-temporal analysis revealed a branched trajectory with two major branches, that is, cell fate 1 and cell fate 2, which could be further divided into 5 cellular states of pericytes (S1-S5). S4 constituted the most of the prebranch, which represented the cellular state of the initial pericyte phenotype. S5 constituted the most of the cell fate 1 branch, which represented the early differentiation state of the pericyte phenotype. S1, S2, S3 constituted the most of the cell fate 2 branch. S3 represented the intermediate differentiation state of the pericyte phenotype, while S1 and S2 represented the terminal differentiation states of the pericyte phenotype. Compared with the uniform distribution of various differentiation states of pericytes in normal skin, the keloid pericytes mainly distributed in the prebranch (S4), cell fate 1 (S5) and the first half of cell fate 2 (S3), representing cellular states of the initial, early and intermediate phases of the pericyte phenotype. Branched expression analysis modeling revealed the overexpression of SOX4, COL4A1, COL6A3, AHR, CXCL3 and IL1R1 genes, et cetera. On the other hand, the localized scleroderma pericytes mainly distributed in the bottom half of cell fate 2 (S1, S2), representing the final differentiated phase of pericyte phenotype, which overexpressed ACTA2 and MYH11 genes.Conclusion:Pericytes in keloid and scleroderma are heterogenous and have different differentiation trajectories. Pericytes in keloid have stem-like characteristics, and play an important role in the pathologic characteristics of invasiveness and recurrence through high expression of genes related to cell stemness, epithelial-mesenchymal transition, invasiveness, and immune microenvironment regulation. However, pericytes in localized scleroderma may mainly transdifferentiate into myofibroblasts, leading to their fibrotic pathological phenotype.

Objective To explore the effect of the combination of Tuina and treadmill training on nerve regeneration after sciatic nerve transection.Methods A total of 60 Sprague-Dawley rats were randomly divided into control group,model group and treatment group equal-ly.The sciatic nerve was transected in the latter two groups and the epineurium suture was delayed four days.Then the treatment group ac-cepted Tuina and treadmill training once a day.The motor conduction velocity(MCV)of sciatic nerve was measured one and two months af-ter intervention in ten rats respectively,while the number of regenerated axons and Schwann cells was counted,and structures of sciatic nev-er were observed under electron microscopy. Results Compared with the model group, the MCV accelerated in the treatment group (P<0.05),the number of axons was not significantly different(P>0.05),and the number of Schwann cells increased two months after modeling (P<0.05),with less injury under electron microscopy.Conclusion Tuina combined with treadmill training can promote the regeneration of the injured peripheral nerve.

ObjectiveTo explore the structural characteristics and functional differences of intestinal flora in patients with type 2 diabetes mellitus （T2DM） of dampness heat trapping spleen（DHTS） syndrome and Qi-Yin deficiency（QYD） syndrome. MethodFrom June 2018 to January 2020，62 T2DM patients with DHTS syndrome and 60 with QYD syndrome were selected from Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine. Serum and fecal samples were collected to compare body mass index（BMI），glucose and lipid metabolism，fasting insulin （FINS） and fasting C-peptide （FCP） levels，and homeostasis model assessment of insulin resistance（HOMA-IR） of the two syndrome types. Fecal samples were extracted for DNA database construction，and 16S rDNA high-throughput sequencing was used to analyze and compare the intestinal flora and metabolic pathways. Result① The BMI，fasting plasma glucose（FPG），2-hour postprandial blood glucose （2 h PBG），total cholesterol（TC），triglyceride（TG），low density lipoprotein（LDL），FINS，FCP，and HOMA-IR were higher in patients with DHTS syndrome than in patients with QYD syndrome，and the high density lipoprotein（HDL） of the former was lower than that of the latter，（P<0.05，P<0.01）. ② In terms of species composition and differences，Bacteroidetes， Clostridia and Gammaproteobacteria were dominant at the class level，and the relative abundance of Clostridia，Mollicutes and Verrucomicrobiae in QYD syndrome group was higher than that in DHTS syndrome group. At the order level，Bacteroidales，Clostridiales and Enterobacteriales were mainly found. The relative abundance of Clostridiales，Erysipelotrichales and Verrucomicrobiales in QYD syndrome group was obviously higher than that in DHTS syndrome group，while Aeromonadales in the former was lower than that in the latter （P<0.05）. At the family level，Bacteroidaceae，Prevotellaceae and Ruminococcaceae were predominant. The relative abundance of Ruminococcaceae，Porphyromonadaceae and Erysipelotrichaceae in QYD syndrome group was higher than that in DHTS syndrome group（P<0.05）. At the genus level，Bacteroides，Prevotella and Parabacteroides were mainly found. The relative abundance of Parabacteroides，Butyrivibrio and Ruminiclostridium in QYD syndrome group was higher than that in DHTS syndrome group，while that of Klebsiella and Megasphaera in DHTS syndrome group was higher than that in QYD syndrome group（P<0.05）. ③ Through Venn analysis of operational taxonomic units（OTU），it was found that there were 49 OTUs in patients with DHTS syndrome patients and 47 OTUs in QYD syndrome patients. ④ The results of OTU β diversity and α analysis showed that Shannon and Simpson indexes had statistical differences，while Ace and Chao indexes had no statistical differences. The intestinal microbial diversity of patients with QYD syndrome was higher than that of patients with DHTS syndrome（P<0.05）. The analysis of similarities （ANOSIM） showed that the difference of β diversity between the two groups was significant（P<0.05）. ⑤ Linear discriminant analysis Effect Size（LEfSe） results demonstrated that Klebsiella，Megasphaera and Aeromonadales could be selected as the key biomarkers for DHTS syndrome； 14 bacteria such as Ruminiclostridium，Burkholderiaceae，Lautropia，Butyrivibrio，Erysipelotrichales can be selected as the key biomarkers for QYD syndrome. ⑥Functional annotation and analysis showed that the DHTS syndrome involved 9 metabolic pathways，including arginine and proline metabolism，lipopolysaccharide biosynthesis，nicotinic acid and nicotinamide metabolism，while the QYD syndrome involved 10 metabolic pathways，including acarbose and valinomycin biosynthesis，glucagon signaling pathway and NOD-like receptor signaling pathway. ConclusionThere are obvious differences in intestinal flora and functions in T2DM patients of DHTS syndrome and QYD syndrome，which can be used as reference for traditional Chinese medicine （TCM） syndrome differentiation and the target of TCM treatment.

Objective To explore the effect of the combination of Tuina and treadmill training on denervation skeletal muscle atrophy. Methods A total of 80 Sprague-Dawley rats (one month old) were randomly divided into control group (n=40) and manipulation group (n=40). Their sciatic nerves were transected, and the manipulative group accepted treadmill training and kneading of Tuina, while the control group accepted no intervention. Their muscle wet weight ratio, muscle satellite cells and insulin-like growth factor I (IGF-I) positive cells count were measured, and HE staining of gastrocnemius muscle were observed one, two, three and four months after intervention, ten rats in each group. Results Compared with the control group, the muscle wet weight ratio decreased three months after intervention (F=4.590, P<0.05), muscle satellite cells increased three months after intervention (F=12.466, P<0.01), and IGF-I positive cells increased two, three and four months after intervention (F>6.489, P<0.05). HE staining showed the skeletal muscle injury relieved somehow.Conclusion The combination of Tuina and treadmill training can relieve denervation skeletal muscle injury, but it is not enough for skeletal muscle atrophy, which may associate with promoting the expression of muscle satellite cells and IGF-I.