OBJECTIVE: To observe the effects of Tongdu Tiaoshen acupuncture (acupuncture for unblocking the obstruction in the governor vessel and regulating the spirit) on the depression-like behavior and the hippocampal neuronal ferroptosis mediated by solute carrier family 7 member 11 (SLC7A11)/glutathione peroxidase 4 (GPX4) pathway in depression rats, and explore the mechanism of this therapy for depression. METHODS: Of 30 male SD rats of SPF grade, 24 rats were selected. According to the random number table, they were divided into a normal group (n=8) and a modeling group (n=16). The rats in the modeling group were subjected to chronic unpredictable mild stress (CUMS) for 28 consecutive days to establish depression model. After modeling, 16 successfully-modeled rats were randomly divided into a model group and an acupuncture group, 8 rats in each one. In the acupuncture group, Tongdu Tiaoshen acupuncture was applied to "Dazhui"(GV14), "Shuigou" (GV26), "Baihui" (GV20) and "Shenting" (GV24). This intervention measure was deliveredonce a day, continuously for 6 days. The intervention discontinued on day 7, and was completed in 4 weeks. Before and after modeling, and after intervention completion, the behavioristics detection was performed using sucrose preference experiment and open field experiment. After intervention, using hematoxylin-eosin (HE) and Nissl staining, the morphology of hippocampal neurons was observed; with Western blot method, the protein expression of GPX4, SLC7A11, Ferritin and acyl-CoA synthetase long-chain family 4 (ACSL4) in hippocampal tissues was detected; with the real-time fluorescence quantitative PCR adopted, the mRNA expression of GPX4, SLC7A11, Ferritin and ACSL4 was detected; and using colorimetry, the hippocampal iron content was determined. RESULTS: After modeling, the sucrose preference rates, the total distance of movement, the standing times and the boxes of horizontal crossing in the model group and the acupuncture group were lower than those in the normal group (P<0.01). After the intervention, the sucrose preference rates, the total distance of movement, the standing times and the boxes of horizontal crossing in the acupuncture group were higher than those in the model group (P<0.01, P<0.05). Compared with the normal group, the number of necrotic cells increased and the number of Nissl bodies decreased in the model group; and when compared with the model group, the neuronal pyknosis and necrosis were ameliorated, the cells were arranged more regularly, the neuronal structure was clear, the matrix was dense, the blood vessels were enriched and the number of Nissl bodies increased in the acupuncture group. In comparison with the normal group, the relative expression of protein and mRNA of hippocampal GPX4, SLC7A11 decreased (P<0.01), it increased in the expression of hippocampal Ferritin and ACSL4 (P<0.01) in the model group. When compared with the model group, in the acupuncture group, the relative expression of protein and mRNA of hippocampal GPX4, SLC7A11 was elevated (P<0.01, P<0.05), it was dropped for hippocampal Ferritin and ACSL4 (P<0.01). In the model group, the hippocampal iron content was elevated when compared with that in the normal group (P<0.01); and it was reduced in the acupuncture group when compared with that in the model group (P<0.05). CONCLUSION Tongdu Tiaoshen acupuncture attenuates depression-like behaviors in the depression rats, which may be related to regulating SLC7A11/GPX4 pathway and inhibiting neuronal ferroptosis in the hippocampus.
Animals ; Ferroptosis ; Male ; Hippocampus/cytology* ; Rats, Sprague-Dawley ; Rats ; Depression/enzymology* ; Phospholipid Hydroperoxide Glutathione Peroxidase/genetics* ; Acupuncture Therapy ; Neurons/metabolism* ; Humans ; Acupuncture Points ; Amino Acid Transport System y+/genetics* ; Glutathione Peroxidase/genetics*

Objective:Based on value orientation,it aimed to scientifically define the concept and connotation of accessibility to novel anticancer drugs,in order to deeply understand the nature and current status of the accessibility issues of novel anticancer drugs,and to provide a reference for the formulation and optimization of policies related to novel anticancer drugs.Methods:Data was collected through literature review and expert interviews,and the concept of drug accessibility was defined using the atomic diagram method.Results:The core images include"affordability","availability","high quality"and"patients".The concept of accessibility to novel anticancer drugs is defined as"the process of ensuring the sustainable supply,equitable access,affordability,and rational use of high-quality anticancer drugs to safeguard the realization of patient benefit goals."The connotation of the value orientation in policies on the accessibility of novel anticancer drugs is profoundly reflected in the multi-dimensional value-driven approach to ensure the ultimate benefit of patients,which includes quality,sustainability,equity,affordability,and rational use.Conclusion:The proposal of the concept and connotation of accessibility provides a theoretical basis for a deep understanding of the accessibility of novel anticancer drugs and offers valuable references for subsequent policy-making and practical operations.

Objective:Based on value orientation,it aimed to scientifically define the concept and connotation of accessibility to novel anticancer drugs,in order to deeply understand the nature and current status of the accessibility issues of novel anticancer drugs,and to provide a reference for the formulation and optimization of policies related to novel anticancer drugs.Methods:Data was collected through literature review and expert interviews,and the concept of drug accessibility was defined using the atomic diagram method.Results:The core images include"affordability","availability","high quality"and"patients".The concept of accessibility to novel anticancer drugs is defined as"the process of ensuring the sustainable supply,equitable access,affordability,and rational use of high-quality anticancer drugs to safeguard the realization of patient benefit goals."The connotation of the value orientation in policies on the accessibility of novel anticancer drugs is profoundly reflected in the multi-dimensional value-driven approach to ensure the ultimate benefit of patients,which includes quality,sustainability,equity,affordability,and rational use.Conclusion:The proposal of the concept and connotation of accessibility provides a theoretical basis for a deep understanding of the accessibility of novel anticancer drugs and offers valuable references for subsequent policy-making and practical operations.

Objective:To investigate the relationship between prolonged PR interval and carotid atherosclerosis(CAS)in middle-aged and elderly patients with type 2 diabetes mellitus(T2DM).Methods:A total of 537 middle-aged and elderly inpatients with T2DM in the Southern Branch of the Sixth People′s Hospital of Shanghai Jiaotong University from January 2019 to January 2021 were selected as the research objects. Color Doppler ultrasound was used to detect bilateral carotid artery intima-media thickness(CIMT). The subjects were divided into carotid atherosclerosis group(CAS group, n=352)and non-carotid atherosclerosis group(NCAS group, n=185). The difference in the PR interval of ECG between the two groups was compared. Pearson or Spearman rank correlation analysis was used for evaluating the correlation of PR interval and CAS lesions with various clinical index. The relationship between PR interval and CAS lesions was adopted by multivariate logistic regression analysis. Results:The average PR interval of middle-aged and elderly patients with T2DM was(164.57±23.02)ms. The average PR interval in CAS group was significantly higher than that in NCAS group [(169.76±24.28) vs (154.70±16.42)ms, P<0.01]. The results of multifactorial logistic regression analysis showed that age, low density lipoprotein-cholesterol, serum osteocalcin, and PR interval were independent factors influencing the development of CAS lesions in middle-aged and elderly patients with T2DM( OR=1.079, 1.936, 0.879, 1.039, respectively, P<0.05 or P<0.01)where each 1 ms increase in PR interval was associated with a 3.9% increase in the risk of CAS in middle-aged and elderly patients with T2DM( OR=1.039, 95% CI 1.006-1.073, P=0.020). Multivariate logistic regression analysis showed that middle-aged and elderly type 2 diabetic patients with PR interval≥158 ms were 4.072 times more likely to have CAS lesions than those with PR interval<158 ms( OR=4.072, 95% CI 1.417-11.702, P<0.01). Conclusion:The PR interval of electrocardiogram is correlated with CAS lesions in middle-aged and elderly patients with T2DM. Middle-aged and elderly type 2 diabetic patients with significantly prolonged PR interval should be reminded to screen for CAS lesions early.

Objective:To explore the evolution of CT characteristics of the "reversed halo sign" of pulmonary tuberculosis, and to further improve the recognition of its CT signs.Methods:Clinical and CT data of 12 patients with pulmonary tuberculosis who were clinically and pathologically confirmed and accompanied with CT manifestation of "reversed halo sign" in First Affiliated Hospital of Henan University of Science and Technology from August 2013 to April 2020 were analyzed retrospectively. Pathological and imaging contrastl analysis was performed on 1 patient undergoing surgical treatment.Results:Among 12 cases with "reversed halo sign", there were 2 cases with single lesion in unilateral lung, 2 cases with multiple lesions in unilateral lung, and 8 cases with multiple lesions in bilateral lungs. Three cases showed only "reversed halo sign", 9 cases showed both halo-like sign and uniform fireworks sign. "Tree-in-bud "sign was found in all 12 patients in the outer ring of the "reversed halo sign". Eight patients received three or more CT examinations, and six of them showed reduction of density and volume of the "reversed halo sign" after standardized anti-tuberculosis treatment. Under the natural course of the disease in two cases, the overall enlargement of the lesion was observed in 1 case, and the overall density of the lesion was reduced and the outer ring wall of the "reversed halo sign" was thinned in 1 case. The pathology of one case after surgical lobectomy showed granulomatous inflammatory nodules of varying sizes containing Langerhans nodule giant cells in the lung parenchyma. The typical caseous necrotic granulomatous nodules were rare here. The "reversed halo sign" showed dense Langerhans nodules in the outer ring, sparse central areas with fibrous hyperplasia and alveolar wall thickening.Conclusions:The outer ring of "reversed halo sign" of pulmonary tuberculosis shows as "tree-in-bud" sign, and its center shows as the fine reticulation pattern. After effective anti-tuberculosis treatment, both the overall density of "reversed halo sign" and the lesion size reduced. Finally, the lesions mostly present as as fine grid shadows for a long time.

Objective: To investigate the effect of miR-124 on the invasion and migration of esophageal cancer KYSE170 cells by regulating autophagy. Methods: miR-124 mimic was transfected into esophageal cancer KYSE170 cells. Transwell assay was used to detect the change of invasion and migration ability of cells. Dual luciferase reporter gene assay was used to verify the targeted regulation of BECN1 (Beclin1) by miR-124, and Western blotting was used to analyze the expressions of BECN1, P62 and LC3 protein. siRNA targeting BECN1 was transfeted into KYSE170 cells, and then the cell invasion and migration ability was calculated by Transwell assay. The expressions of BECN1, P62 and LC3 protein were detected by Western blotting. miR-124 mimic and BECN1 over-expression plasmid were co-transfected into KYSE170 cells, and then Transwell assay was used to detect the changes of cell invasion and migration ability, and Western blotting to examine the expression levels of autophagy-related gene. Results: The invasion and migration ability of KYSE170 cells were significantly inhibited after transfection with miR-124 mimic (All P<0.05). The expression of autophagyrelated protein P62 was increased, and the expression of BECN1 and LC3 was significantly decreased (All P<0.01); in addition, the activity of luciferase reporter gene was also significantly reduced (P<0.01). Silencing BECN1 expression inhibited the invasion and migration of esophageal cancer KYSE170 cells (P<0.01). However, after co-transfection with BECN1 over-expression plasmids, the effects of miR-124 mimic on the autophagy, invasion and migration of esophageal carcinoma KYSE170 cells were significantly weakened (P<0.01), it was also accompanied with lower P62 expression, and higher LC3 expression (P<0.01). Conclusion: miR-124 mimic can inhibit the invasion and migration of esophageal carcinoma cells. The mechanism may be related to the autophagy-related gene BECN1 expression.

Objective: To evaluate the expression of melanoma antigen A family（MAGE-As）in the peripheral blood of patients with esophageal carcinoma (EC), and to analyze its correlations to the clinicopathological features and the prognosis of EC patients. Methods: mRNA expression of MAGE-As in peripheral blood from 153 EC patients and 30 healthy donors was detected using multiplex semi-nested PCR. In addition, restriction endonuclease treatment was used to determine the expression of MAGE-As family members, including MAGE-A1, A2, A3, A4 and A6. Results: The positive expression of MAGE-As was observed in 30 of 153 EC patients (19.61%) in peripheral blood. The positive expression rate of MAGE-A1, A2, A3, A4, A6 was 10.46% (16/153), 16.34%(25/153), 9.8% (15/153), 11.11% (17/153) and 18.30%(28/153), respectively. Additionally, the expression of MAGE-As was positively associated with clinical stage, lymphatic metastasis and distant metastasis (all P<0.05). The positive expressions of MAGE-As and its sub-type genes were all associated with low 5-year overall survival of ES patients (all P<0.05). Expression of MAGE-As, tumor volume, lymphatic metastasis and distant metastasis can be used as independent prognostic factors for the survival of EC patients (all P<0.01). Conclusion: The expression of MAGE-As in peripheral blood of EC patients was associated with the prognosis of EC, and may be used as an important indicator for the prognosis of esophageal carcinoma.

Objective: To detect the expression of miR-133a-3p in gastric cancer (GC) tissues and plasma of GC patients, and to investigate its effect on the proliferation of GC cells as well as its correlation toprognosis of GC patients. Methods: 52 cases of cancertissues (non-necrosis part) and corresponding adjacent tissues as well as the pre-operative peripheral blood samples from GC patients, who underwent surgery at Department of General Surgery, the Forth Hospital of Hebei Medical University(Shijiazhuang, China) between May 2012 and May 2013, were collected for this study. The plasma sample (n=35) from healthy donors were obtained during their physical examination. RT-qPCR was adopted to detect the expression of miR-133a-3p in gastric cancer tissues, adjacent tissuesand plasma samples of GC patients and healthy volunteers. The relationships between miR-133a-3p expression and the median DFS as well as clinicopathological parameters were also analyzed. CCK-8 assay was adopted to detect the effect of miR-133a-3p silence or over-expression on proliferation of gastric cancer SGC7901 cells. Results: miR-133a-3p was dramatically decreased in gastric cancer tissues (P<0.01), and its expression was associated with TNM stage, tumor infiltration (T), lynphonode metastasis (N), and vascular tumor thrombus (all P<0.01); miR-133a-3p was significantly increased in the plasma of GC patients (P<0.01), and its expression was associated with TNM stage, lynphonode metastasis (N), and vascular tumor thrombus (all P<0.05). miR-133a-3p expression was positively correlated with serum CA199 level of GC patients (P<0.01). The median DFS of patients with high miR-133a-3pexpression in cancer tissues was significantly longer than that of the patients with low expression(20.8 vs 14.8 months, P<0.05); The median DFS of patients with high plasma miR-133a-3p expression was significantly shorter than that of the patients with low expression (14.4 vs 20.3 months, P<0.05). Over-expression of miR-133a-3p could significantly inhibit the proliferation of gastric cancer SGC7901 cells, while miR-133a-3p silence could significantly promote the proliferation (all P<0.05). Conclusion: miR-133a-3p could significantlyinhibit the proliferation of SGC7901 cells; miR-133a-3p aberrantlyexpressed in gastric cancer tissues and plasma, and obviously correlated with prognosis of gastric cancer patients, which may be used as a potential clinical bio-maker for early diagnosis and treatment as well as the prognosis prediction of gastric cancer.

[Abstract] Objective: To investigate the regulating effects of miR-92b on the expression of EZH2 (enhancer of zeste homolog 2) gene and the proliferation and invasion abilities of esophageal cancer (EC) cells. Methods: Fifteen cases of esophageal cancer tissues that preserved in the research center of the Fourth HospitalAffiliated to Heibei Medical University from January 2016 to January 2017 were selected for this study. The bioinformatics tool was used to predict the possible miRNAs that might target EZH2. The mimics of predicted miRNAs were transfected into human esophageal carcinoma cell lines Eca109, respectively. Then the regulation effect of miRNAs on EZH2 gene expression was validated by real-time PCR, Western blotting and dual luciferase reporter experiment. In the meanwhile, EZH2 over-expression plasmids were co-transfected into esophageal carcinoma Eca109 cells, and the effects of miRNAs and EZH2 expression changes on the proliferation, apoptosis , invasion and migration of esophageal carcinoma cells were detected by CCK-8 method, Flow Cytometry, Transwell Invasion and migration assay, respectively. Results: Bioinformatics analysis showed that miR-92b, let7a and miR-25 could combine with potential binding sites at 3’-terminal non-translation region of EZH2 gene. Real-time PCR results showed that only miR-92b was able to regulate the expression of EZH2, and miR-92b was negatively correlated to EZH2 in esophageal cancer (P<0.01). Compared with mimic-NC, the expression of EZH2 mRNA, protein and luciferase activity in Eca109 cells after miR-92b mimic transfection was significantly down-regulated (both P<0.01). However, miR-92b mimic transfection had no effect on the apoptosis of Eca109 cells. Moreover, the proliferation, invasion and migration of Eca109 cells were significantly inhibited after transfection with miR-92b-mimic (P<0.01). In addition, after co-transfection with EZH2 over-expression plasmids, the effects of miR-92b-mimic on the proliferation, invasion and migration of Eca109 cells were significantly weakened (P<0.01). Conclusion: miR-92b can inhibit the proliferation,invasionandmigrationofesophagealcarcinomacells,anditsmechanismmayberelatedtoitstargetregulationofEZH2. ··

Objective: To screen related genes of melanoma-associated antigen-A11 (MAGE-A11) in breast cancer cells based on highthroughput DNAmicroarray technology, and to validate from the aspects of quantity and function. Methods: DNAmicroarray was used to screen the differently-expresseddown-stream mRNAs of MAGE-A11 in breast cancercelllines (MCF-7, MDA-MB-231 and BT-549). Cluster analysis was applied on representative genes and quantitative RT-PCR was used to validate. CCK-8, scratch wound healing assay and Transwell assaywere used to detect the effect of MAGE-A11 on the proliferation,migration and invasion of breast cancer cells. Results: Over-expression of MAGE-A11 caused the differential expression of 1608 down-stream genes in 3 breast cancer cell lines, which was associated with various cell functions such as protein ubiquitination,cell proliferation and apoptosis, tumor invasion and metastasis.qRT-PCR validated that the expression of ZNF-451, CENPTJ, CDK13, API5 and LMO7, which were highly expressed in microarray, were also significantly higher than those in control group (P<0.01);in addition, SHPRH, PML, MARK2, LIMA1 and ANGPTL4, which were low-expressed in microarray, were also significantly lower than those in control group (P<0.01). MAGE-A11transfection directly increased the proliferation of breast cancer MCF-7, MDA-MB-231 and BT-549 cells at 72 h (all P<0.01); compared with control group after transfectionexhibited obvious wound healing at 48 h (P<0.05 or P<0.01) and significantly increased trans-membrane cell numbers (all P<0.01). Conclusion: Many differentially expressed genes related to ubiquitination, cell proliferation and apoptosis, tumor invasion and migration were screened in MCF-7, MDA-MB-231 and BT-549 breast cancer cells. Among them, 10 typical differentially expressed genes were identified in terms of quantity and function.