Objective:Exploration on the mechanism of acupuncture-targeted therapy for rheumatoid arthritis and identification of candidate Chinese materia medica.Methods:Targets of acupuncture for RA were collected from related literature, and GSE205962 gene chips were obtained from GEO database. The weighted co-expression network (WGCNA) was constructed by screening differentially expressed genes using R 4.3.1. The collection targets were taken to intersect with the differential genes, and the common genes were obtained for high-throughput GO and KEGG enrichment. The Hub genes were screened by constructing protein interaction networks using the String database and Cytoscape 3.9.0 software. The CIBERSORT algorithm was used to explore the mechanisms of RA immune infiltration, and finally the Coremine Medical database was applied to predict Chinese materia medica.Results:A total of 8 key targets were selected (RHOA, JAK1, FYN, LCK, STAT1, STAT3, EGF, GATA3). The results of immune infiltration analysis showed that various immune cells were closely related to the biological processes of rheumatoid arthritis, including γδ T cells, monocytes, regulatory T cells, CD8 T cells, etc. Saposheikovize Radix, Cinnamomi Ramulus, Anemarrhenae Rhizoma, Dendrobii Caulis, Rehmannine Radix, Lonicerae Japonicae Flos, Pinelliae Rhizoma, Cimicifugae Rhizoma, Poria, Stephaniae Tetrandrae Radix were the 10 kinds of commonly used key Chinese materia medica in the treatment of RA.Conclusions:Acupuncture and moxibustion may treat RA by regulating RHOA, JAK1, FYN, LCK and other core targets. This study predicts that Chinese materia medica can be used as a source of drugs for the treatment of RA, and can provide new ideas for the combination of acupuncture and medicine for the treatment of RA.

Objective:To analyze the clinical efficacy and safety of camrelizumab combined with apatinib as a second-line therapy for unresectable hepatocellular carcinoma (HCC).Methods:Ninety-four cases with mid-and advanced-stage HCC who received camrelizumab combined with apatinib as second-line treatment were enrolled. Routine blood test, blood biochemical indexes, tumor stage, tumor imaging characteristics, previous treatment strategies and other clinical data before treatment were documented. Imaging examination follow-up results and adverse reactions during treatment were followed up until the end of follow-up or loss of follow-up or death. Kaplan-Meier method was used to analyze the clinical efficacy.Results:As of the last follow-up, 94 cases with mid-and advanced-stage HCC had received camrelizumab combined with apatinib as second-line treatment. Among them, 15 cases were lost to follow-up, 31 cases died, and 48 cases survived. The overall remission rate was 31.9%. The overall disease control rate was 71.3%. The median time to disease-free progression was 6.6 months. The median time to disease progression was not yet available. The 1-year cumulative survival rate was 62.3%. Grade 3 and above adverse reactions mainly included were thrombocytopenia (7.4%), abdominal pain (4.3%), active hepatitis (4.3%), leukopenia (4.3%), diarrhea (3.2%), hand-foot syndrome (3.2%). All adverse reactions were effectively controlled.Conclusion:Camrelizumab combined with apatinib can effectively prolong the survival period of patients with mid-and advanced-stage HCC, and it is well tolerated.

Corneal collagen cross-linking (CXL) has been proved to be a very safe and effective technique to halt the progression of many cornea diseases, such as ectasia disease, infectious keratitis and ametropia.CXL techniques include conventional CXL, accelerated CXL, transepithelial CXL and photochemical CXL.CXL can prevent the progression of dilated keratopathy, postpone or avoid corneal transplant surgery, and has an auxiliary function in the treatment of bullous keratopathy, infectious keratitis except herpes virus infection, and ametropia.CXL is not effective for patients with ocular herpes virus infection and patients with thin cornea and may even worsen the development of the disease.This review summarized the advantages and disadvantages of common CXL techniques, indications, contraindications, and complications, which may help enhancing the clinical application of CXL and reducing its complications.

Objective:To evaluate the effect of mushroom-like pattern femtosecond laser enabled penetrating keratoplasty with different laser energies on the ring incision and corneal endothelial cells, and compare with conventional penetrating keratoplasty.Methods:According to the point/line separation and blasting energy, 48 porcine corneas were randomly divided into 6 groups by using a random number table.In group A, B, C, D and E, the point/line separation was 4/4, 4/4, 8/8, 8/8 and 4/2 μm, and the blasting energy was 1.5, 2.0, 1.5, 2.0 and 2.0 μJ; In group F, a negative pressure ring drill was used to make penetrating incisions.There were 8 porcine corneas in each group.Femtosecond laser of 200 kHz was used to make corresponding mushroom-shaped penetrating incisions on the porcine corneas, and compared with porcine corneas incisions with a ring drill.The optical and scanning electron microscope images were used to evaluate the glossiness of ring incisions.Four human corneas used femtosecond laser for mushroom-like penetrating incisions with point/line separation 8/8 μm, and blasting energy 2.0 μJ as the experimental group; three human corneas received a negative pressure ring drill for penetrating incisions as the control group.The loss rate of corneal endothelial cells was observed and compared between experimental group and control group.The donor guardians agreed to the acquisition and use of the donors' cornea and signed informed consents.This study protocol was approved by the Ethics Committee of Nanjing First Hospital.Results:Femtosecond laser mushroom penetrating ring incision completion rate was 100%.The corneal ring incision in each group was stepped, and sections of ring incisions in group A and group E were glossiest by the light microscope.Sections of ring incisions in group E were glossiest by the laser scanning electron microscope.The overall glossiness scores of the corneal ring incisions among various groups were statistically significantly different by the light microscopy ( F=22.75, P<0.01). Among them, the glossiness scores in the group A were higher than those in the group B, and the glossiness scores in the group C were higher than those in the group D, with statistically significant differences (both P<0.05). The overall glossiness scores of corneal ring incisions among various groups were statistically significantly different by laser scanning electron microscopey ( F=122.33, P<0.01). Among them, the glossiness scores in the group A were higher than those in the group B, and the the glossiness scores in the group C were higher than those in the group D, with statistically significant differences (both P<0.05). The corneal endothelial cells showed regular shape and tight arrangement in the experimental group, and irregular shape and loose connections were seen in the control group.The average corneal endothelial cell loss rate in the experimental group was (2.2±1.3)%, lower than (6.7±2.1)% of the control group, with a significant difference between them ( t=3.569, P<0.05). Conclusions:Femtosecond laser can produce perfect mushroom configuration, and the ring incision glossiness is better in comparison with trephine cutting.Femtosecond laser ring cut can lessen corneal endothelial loss.

ObjectiveTo explore the intervention mechanism of Xiangsha Liujunzi Tang in rats with functional dyspepsia （FD） based on the Ras homolog gene family member A （RhoA）/Rho-associated coiled-coil containing protein kinase 2 （ROCK2）/Myosin phosphatase target Subunit 1 （MYPT1） pathway. MethodSixty male SD suckling rats in SPF grades were randomly divided into blank group （n=10） and model group （n=50）. The comprehensive modeling method （gavage administration of iodoacetamide+exhaustion of swimming+disturbance of hunger and satiety） was used to replicate the rat model of FD. After successful replication of the model， the rats in the model group were randomly divided into model group， mosapride group， and high， middle， and low-dose Xiangsha Liujunzi Tang groups， with 10 rats in each group. Rats in the blank group and model group were given 10 mL kg^-1·d^-1 normal saline， those in the mosapride group were given 1.35 mg·kg^-1·d^-1 mosapride， and those in the high， middle， and low-dose Xiangsha Liujunzi Tang groups were given 12， 6， and 3 g·kg^-1·d^-1 Xiangsha Liujunzi Tang， respectively. The intervention lasted 14 days. The general living conditions of rats were observed before and after modeling and administration， and the 3-hour food intake and body mass of rats were measured. After intervention， the intestinal propulsion rate of rats was measured， and the pathological changes in the gastric tissue were observed by hematoxylin-eosin （HE） staining. The content of choline acetyl transferase （ChAT） and vasoactive intestinal peptide （VIP） in the medulla oblongata and gastric tissue homogenate was determined by enzyme-linked immunosorbent assay （ELISA）， the distribution of adenosine triphosphate （ATP） enzyme in gastric antrum smooth muscle was observed by frozen section staining， and the protein expression levels of RhoA， ROCK2， and phosphorylated-myosin phosphatase target subunit 1 （p-MYPT1） in the gastric tissue were detected by Western blot. ResultCompared with the blank group， the model group had withered hair， lazy movement， slow action， poor general living condition， lower 3-hour food intake， body mass， and lower intestinal propulsion rate （P<0.05）， whereas no obvious abnormality in gastric histopathology. In the model group， the content of ChAT in the medulla oblongata and gastric tissue decreased， the content of VIP in gastric tissue increased， the distribution of ATP enzyme in gastric antrum smooth muscle decreased significantly， and the protein expression levels of RhoA， ROCK2， and p-MYPT1 in the gastric tissue decreased significantly （P<0.05）. As compared with the model group， the general living condition of rats in each intervention group was significantly improved， and the 3-hour food intake， body mass， and intestinal propulsion rate were significantly increased （P<0.05）. There was no significant difference in gastric pathology in the intervention groups. The content of ChAT in the medulla oblongata and gastric tissue increased significantly， the content of VIP in the gastric tissue decreased， the distribution of ATP enzyme in gastric antrum smooth muscle increased significantly， and the protein expression levels of RhoA， ROCK2， and p-MYPT1 in the gastric tissue increased significantly （P<0.05）. The intervention effect of Xiangsha Liujunzi Tang group on the above indexes was dose-dependent. ConclusionXiangsha Liujunzi Tang can effectively improve the general living condition and gastric motility of rats with FD， and its specific mechanism may be related to the activation of the RhoA/ROCK2/MYPT1 pathway in the gastric tissue to regulate smooth muscle relaxation and contraction and promote gastric motility.

OBJECTIVE: To investigate the mechanism by which epigallocatechin gallate (EGCG) induces gene demethylation and promotes the apoptosis of acute myeloid leukemia KG-1 and THP-1 cell lines. METHODS: KG-1 and THP-1 cells treated with 25, 50, 75, 100 or 150 μg/mL EGCG for 48 h were examined for gene methylation using MSP and for cell proliferation using MTT assay. The changes in cell cycle and apoptosis of the two cell lines after treatment with EGCG for 48 h were detected using flow cytometry. The mRNA and protein expressions of DNMT1, CHD5, p19, p53 and p21 in the cells were detected using RT-quantitative PCR and Western blot. RESULTS: EGCG dose-dependently reversed hypermethylation of gene and reduced the cell viability in both KG-1 and THP-1 cells ( < 0.05). EGCG treatment caused obvious cell cycle arrest in G1 phase, significantly increased cell apoptosis, downregulated the expression of DNMT1 and upregulated the expressions of CHD5, p19, p53 and p21 in KG-1 and THP-1 cells ( < 0.05). CONCLUSIONS EGCG reduces hypermethylation of gene in KG-1 and THP-1 cells by downregulating DNMT1 to restore its expression, which results in upregulated expressions of p19, p53 and p21 and induces cell apoptosis.

Objective: To investigate differentially expressed genes and related signaling pathways in patients with dermatomyositis/clinical amyopathic dermatomyositis (DM/CADM) complicated by interstitial lung disease or malignant tumors. Methods: From January 2017 to January 2018, 27 DM/CADM patients were enrolled from Department of Dermatology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, and divided into 3 groups according to the complications: 10 with interstitial lung disease, 8 with malignant tumors, and 9 without interstitial lung disease or malignant tumors. Meanwhile, 7 healthy controls were enrolled into this study. High-throughput RNA sequencing was performed to screen differentially expressed genes in peripheral blood in the above 4 groups. Then, these genes were subjected to gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Results: Compared with the healthy controls, 4 820 up-regulated genes and 137 down-regulated genes were identified in DM/CADM patients; GO analysis revealed 49 significantly enriched items in the DM/CADM patients, 37 (75.5%) of which were associated with biological processes; KEGG analysis showed that differentially expressed genes were mainly enriched in infection-, tumor- and immune-related pathways in DM/CADM patients. Compared with the patients without interstitial lung disease or malignant tumors, 272 up-regulated genes and 158 down-regulated genes were identified in the patients with interstitial lung disease; GO analysis revealed 157 significantly enriched items, 114 (72.6%) of which were associated with biological processes; KEGG analysis showed that differentially expressed genes were mainly enriched in bacterial infection- and autoimmune/inflammatory-related pathways in the patients with interstitial lung disease. Compared with the patients without interstitial lung disease or malignant tumors, 398 up-regulated genes and 68 down-regulated genes were identified in the patients with malignant tumors; GO analysis revealed 117 significantly enriched items, 94 (80.3%) of which were associated with biological processes; KEGG analysis showed that differentially expressed genes were mainly enriched in glycosylation-, metabolism- and tumor-related signaling pathways in the patients with malignant tumors. Conclusions Differences existed in transcriptomes and pathways between the DM/CADM patients and healthy controls, as well as between the patients with interstitial lung disease or malignant tumors and patients without these complications. Bacterial infection- and cytokine/chemokine-related pathways were significantly enriched in the patients with DM/CADM complicated by interstitial lung disease, while those pathways related to glycosylation, protein metabolism, angtigen presentation and cytotoxic effects of natural killer cells were significantly enriched in the patients with DM/CADM complicated by malignant tumors.

Scavenging reactive oxygen species (ROS) by antioxidants is the important therapy to cerebral ischemia-reperfusion injury (CIRI) in stroke. The antioxidant with novel dual-antioxidant mechanism of directly scavenging ROS and indirectly through antioxidant pathway activation may be a promising CIRI therapeutic strategy. In our study, a series of chalcone analogues were designed and synthesized, and multiple potential chalcone analogues with dual antioxidant mechanisms were screened. Among these compounds, the most active not only conferred cytoprotection of HO-induced oxidative damage in PC12 cells through scavenging free radicals directly and activating NRF2/ARE antioxidant pathway at the same time, but also played an important role against ischemia/reperfusion-related brain injury in animals. More importantly, in comparison with mono-antioxidant mechanism compounds, exhibited higher cytoprotective and neuroprotective potential and Overall, our findings showed compound could emerge as a promising anti-ischemic stroke drug candidate and provided novel dual-antioxidant mechanism strategies and concepts for oxidative stress-related diseases treatment.