Increased serum extrachromosomal circular DNA SORBS1circle level is associated with insulin resistance in patients with newly diagnosed type 2 diabetes mellitus.

BACKGROUND: Extrachromosomal circular DNAs (eccDNAs) exist in human blood and somatic cells, and are essential for oncogene plasticity and drug resistance. However, the presence and impact of eccDNAs in type 2 diabetes mellitus (T2DM) remains inadequately understood. METHODS: We purified and sequenced the serum eccDNAs obtained from newly diagnosed T2DM patients and normal control (NC) subjects using Circle-sequencing. We validated the level of a novel circulating eccDNA named sorbin and SH3-domain- containing-1circle97206791-97208025 (SORBS1circle) in 106 newly diagnosed T2DM patients. The relationship between eccDNA SORBS1circle and clinical data was analyzed. Furthermore, we explored the source and expression level of eccDNA SORBS1circle in the high glucose and palmitate (HG/PA)-induced hepatocyte (HepG2 cell) insulin resistance model. RESULTS: A total of 22,543 and 19,195 eccDNAs were found in serum samples obtained from newly diagnosed T2DM patients and NC subjects, respectively. The T2DM patients had a greater distribution of eccDNA on chromosomes 1, 14, 16, 17, 18, 19, 20 and X. Additionally, 598 serum eccDNAs were found to be upregulated, while 856 eccDNAs were downregulated in T2DM patients compared with NC subjects. KEGG analysis demonstrated that the genes carried by eccDNAs were mainly associated with insulin resistance. Moreover, it was validated that the eccDNA SORBS1circle was significantly increased in serum of newly diagnosed T2DM patients (106 T2DM patients vs. 40 NC subjects). The serum eccDNA SORBS1circle content was positively correlated with the levels of glycosylated hemoglobin A1C (HbA1C) and homeostasis model assessment of insulin resistance (HOMA-IR) in T2DM patients. Intracellular eccDNA SORBS1circle expression was significantly enhanced in the high glucose and palmitate (HG/PA)-induced hepatocyte (HepG2 cell) insulin resistance model. Moreover, the upregulation of eccDNA SORBS1circle in the HG/PA-treated HepG2 cells was dependent on generation of apoptotic DNA fragmentation. CONCLUSIONS: These results provide a preliminary understanding of the circulating eccDNA patterns at the early stage of T2DM and suggest that eccDNA SORBS1circle may be involved in the development of insulin resistance.

Sesamin ameliorates nonalcoholic steatohepatitis through inhibiting hepatocyte pyroptosis in vivo and in vitro.

Sesamin (Ses) is a natural lignan abundantly present in sesame and sesame oil. Pyroptosis, a newly identified type of pro-inflammatory programmed necrosis, contributes to the development of non-alcoholic steatohepatitis (NASH) when hepatocyte pyroptosis is excessive. In this study, Ses treatment demonstrated an improvement in hepatic damage in mice with high-fat, high-cholesterol diet-induced NASH and palmitate (PA)-treated mouse primary hepatocytes. Notably, we discovered, for the first time, that Ses could alleviate hepatocyte pyroptosis both in vivo and in vitro. Furthermore, treatment with phorbol myristate acetate, a protein kinase Cδ (PKCδ) agonist, increased PKCδ phosphorylation and attenuated the protective effects of Ses against pyroptosis in PA-treated mouse primary hepatocytes. Mechanistically, Ses treatment alleviated hepatocyte pyroptosis in NASH, which was associated with the regulation of the PKCδ/nod-like receptor family CARD domain-containing protein 4/caspase-1 axis. This study introduces a novel concept and target, suggesting the potential use of functional factors in food to alleviate liver damage caused by NASH.

Sesamin ameliorates nonalcoholic hepatic steatosis by inhibiting CD36-mediated hepatocyte lipid accumulation in vitro and in vivo.

Hepatic steatosis is a critical factor in the development of nonalcoholic steatohepatitis (NASH). Sesamin (Ses), a functional lignan isolated from Sesamum indicum, possesses hypolipidemic, liver-protective, anti-hypertensive, and anti-tumor properties. Ses has been found to improve hepatic steatosis, but the exact mechanisms through which Ses achieves this are not well understood. In this study, we observed the anti-hepatic steatosis effects of Ses in palmitate/oleate (PA/OA)-incubated primary mouse hepatocytes, AML12 hepatocytes, and HepG2 cells, as well as in high-fat, high-cholesterol diet-induced NASH mice. RNA sequencing analysis revealed that cluster of differentiation 36 (CD36), a free fatty acid (FA) transport protein, was involved in the Ses-mediated inhibition of hepatic fat accumulation. Moreover, the overexpression of CD36 significantly increased hepatic steatosis in both Ses-treated PA/OA-incubated HepG2 cells and NASH mice. Furthermore, Ses treatment suppressed insulin-induced de novo lipogenesis in HepG2 cells, which was reversed by CD36 overexpression. Mechanistically, we found that Ses ameliorated NASH by inhibiting CD36-mediated FA uptake and upregulation of lipogenic genes, including FA synthase, stearoyl-CoA desaturase 1, and sterol regulatory element-binding protein 1. The findings of our study provide novel insights into the potential therapeutic applications of Ses in the treatment of NASH.

Metformin improves nonalcoholic fatty liver disease in db/db mice by inhibiting ferroptosis.

Nonalcoholic fatty liver disease (NAFLD) is the primary complication of type 2 diabetes (T2DM)-related liver disease, lacking effective treatment options. Metformin (Met), a widely prescribed anti-hyperglycemic medication, has been found to protect against NAFLD. Ferroptosis, a newly discovered form of cell death, is associated with the development of NAFLD. Despite this association, the extent of Met's protective effects on NAFLD through the modulation of ferroptosis has yet to be thoroughly investigated. In the present study, the administration of erastin or Ras-selective lethal 3 (RSL3), both known ferroptosis inducers, resulted in elevated cell mortality and reduced cell viability in AML12 hepatocytes. Notably, Met treatment demonstrated the capacity to mitigate these effects. Furthermore, we observed increased ferroptosis levels in both AML12 hepatocytes treated with palmitate and oleate (PA/OA) and in the liver tissue of db/db mice. Met treatment demonstrated significant reductions in iron accumulation and lipid-related reactive oxygen species production, simultaneously elevating the glutathione/glutathione disulfide ratio in both PA/OA-treated AML12 hepatocytes and the liver tissue of db/db mice. Interestingly, the anti-ferroptosis effects of Met were significantly reversed with the administration of RSL3, both in vitro and in vivo. Mechanistically, Met treatment regulated the glutathione peroxidase 4/solute carrier family 7 member 11/acyl-CoA synthetase long-chain family member 4 axis to alleviate ferroptosis in NAFLD hepatocytes. Overall, our findings highlight the crucial role of ferroptosis in the development of T2DM-related NAFLD and underscore the potential of Met in modulating key factors associated with ferroptosis in the context of NAFLD.

SOV sensitizes gastric cancer cells to radiation by suppressing radiation-induced autophagy in vitro and in vivo.

Vanadium is a transition metal that naturally occurs in the environment and has a variety of biological and physiological impacts on humans. Sodium orthovanadate (SOV), a well-known chemical compound of vanadium, has shown notable anti-cancer activity in various types of human malignancies. However, the effect of SOV on stomach cancer is yet undetermined. Furthermore, only a few studies have investigated the association of SOV and radiosensitivity with stomach cancer. Our study has investigated the ability of SOV to increase the sensitivity of gastric cancer cells to radiation. To detect autophagy triggered by ionizing radiation and the influence of SOV on cell radiosensitivity, the Cell Counting Kit-8 (CCK8) test, EDU staining experiment, colony formation assay, and immunofluorescence were performed. The possible synergistic effects of SOV and irradiation were examined in vivo using a xenograft mouse model of stomach cancer cells. Both in vitro and in vivo studies showed that SOV markedly reduced the growth of stomach cancer cells and improved their radiosensitivity. Our results showed that SOV increased gastric cancer cells' radiosensitivity, thereby blocking the radiation-induced autophagy-related protein, ATG10. Thus, SOV can be considered a potential agent for radiosensitizing gastric cancer.

Protective effects of metformin on pancreatic ß-cell ferroptosis in type 2 diabetes in vivo.

Metformin (Met) is the recommended first-line therapeutic drug for type 2 diabetes mellitus (T2DM) and exerts protective effects on ß-cell damage. Ferroptosis, a new form of cell death, is associated with pancreatic islet injury in patients with T2DM. However, the protective effects of Met treatment against ß-cell damage through ferroptosis modulation remain under-reported. This study investigated the in vivo effects of Met treatment on pancreatic ß-cell ferroptosis using two different diabetic mouse models, namely, low-dose streptozotocin (STZ) and high-fat diet (HFD)-induced diabetic mice and db/db mice. Met treatment significantly restored insulin release, reduced cell mortality, and decreased the overproduction of lipid-related reactive oxygen species in the islets of both STZ/HFD-induced diabetic mice and db/db mice. Administration of the Ras-selective lethal 3 injection significantly attenuated the antiferroptosis effects of Met. Mechanistically, Met treatment alleviated ß-cell ferroptosis in T2DM, which was associated with the regulation of the GPX4/ACSL4 axis in the islets. In conclusion, our findings highlight the significance of ferroptosis in T2DM ß-cell damage and provide novel insights into the protective effects of Met against islet ß cells.

Metformin alleviates glucolipotoxicity-induced pancreatic ß cell ferroptosis through regulation of the GPX4/ACSL4 axis.

Ferroptosis, a new type of cell death, is associated with pancreatic ß cell damage. However, the role of glucolipotoxicity in inducing ß cell ferroptosis remains unclear. Metformin (Met), exenatide (Exe), and saxagliptin (Sax) are frequently used anti-hyperglycaemic drugs. However, their protective effects on ß cells through ferroptosis modulation are not well-established. In this study, we observed significant ferroptosis in NIT-1 cells and primary mouse islets after exposure to high glucose and palmitate (HG/PA). Compared to Exe and Sax, Met significantly alleviated glucolipotoxicity-induced pancreatic ß cell ferroptosis. Blocking the activity of glutathione peroxidase 4 (GPX4) with Ras-selective lethal 3 or inhibiting its expression by small interfering RNA transfection significantly attenuated the anti-ferroptosis effects of Met. Mechanistically, Met alleviates HG/PA-induced ß cell ferroptosis by regulating the GPX4/ACSL4 axis. Collectively, our findings highlight the significance of ferroptosis in pancreatic ß cell glucolipotoxicity-induced injury and provide novel insights into the protective effects of Met on islet ß cells.

[Acupuncture combined with cranial electrotherapy stimulation on generalized anxiety disorder: a randomized controlled trial].

OBJECTIVE: To observe the therapeutic effect of acupuncture combined with cranial electrotherapy stimulation (CES) on generalized anxiety disorder (GAD). METHODS: A total of 200 patients with GAD were randomized into an acupuncture+CES group, an acupuncture group, a CES group and a medication group, 50 cases in each one. In the medication group, patients were treated with tandospirone citrate tablet orally, 10 mg after breakfast, lunch and dinner respectively. In the CES group, CES was adopted by SCS brain electromedical instrument, 60 min each time, once a day. In the acupuncture group, acupuncture was applied to Baihui (GV 20), Sishencong (EX-HN 1), Yintang (GV 29), Shenting (GV 24), etc., 30 min each time, once a day. In the acupuncture+CES group, CES was adopted before acupuncture. Treatment of sixty days was required in the 4 groups. Before and after treatment, the scores of Hamilton anxiety scale (HAMA), World Health Organization's quality of life questionnaire-brief version (WHOQOL-BREF) and treatment emergent symptom scale (TESS) were observed, the clinical effect was evaluated, and the relapse of anxiety during follow-up of 1 year after treatment was recorded in the 4 groups. RESULTS: Compared before treatment, the scores of HAMA after treatment were decreased (P<0.05), the scores of WHOQOL- BREF after treatment were increased in the 4 groups (P<0.05), and the improvements of above scores in the acupuncture+CES group were greater than the other 3 groups (P<0.05). The score of TESS after treatment and the relapse rate of 1-year follow-up in the medication group were higher than those in the other 3 groups (P<0.05). The total effective rate in the acupuncture+CES group were superior to the other 3 groups (P<0.05). CONCLUSION: Acupuncture combined with CES can effectively relieve the symptoms in patients with GAD, improve the quality of life, reduce the occurrence of adverse reactions and the relapse rate, and its clinical effect is obviously superior to the western medication, the simple application of acupuncture or CES.

[Observation on clinical efficacy of depression treated with the alliance of acupuncture and medication].

OBJECTIVE: To observe the differences in the clinical efficacy on depression in comparison among the alliance of acupuncture and Chinese herbal medicine, simple herbal medicine and flupentixol and melitracen tablets. METHODS: Two hundred and twenty-seven cases were randomized into a Chinese medicine group(75 cases), an acupuncture + medication group(78 cases) and a western medicine group(74 cases). In the Chinese medicine group, Shugan Jianwei Anshen Decoction was prescribed, in which Radix Bupleuri and Radix Paeoniae Alba were the monarch herbs, one dose a day. In the acupuncture + medication group, on the basic treatment of Chinese medicine, acupuncture was combined every day at Baihui (GV 20), Sishencong (EX-HN 1), Shenmen (HT 7), Lingdao (HT 4), Daling (PC 7), Laogong (PC 8), Yongquan (KI 1), etc. In the western medicine group, deanxit was taken orally, 1 tablet after getting up in the morning and after lunch respectively. The treatment lasted for 8 weeks in the three groups. The change of Hamilton depression scale (HAMD) score and clinical efficacy were observed before and after treatment. RESULTS: In 8 weeks of treatment, in the acupuncture + medication group, 19 cases were cured, 35 cases were markedly effective, 14 cases were effective and 10 cases failed, the effective rate was 87.2%; in the western medicine group, 11 cases were cured, 32 cases were markedly effective, 18 cases were effective and 13 cases failed, the effective rate was 82.4%; in the Chinese medicine group, 5 cases were cured, 14 cases were markedly effective, 35 cases were effective and 21 cases failed, the effective rate was 72.0%. HAMD assessment was done in 8 weeks of treatment, which was (5.71 +/- 4.32) scores in the acupuncture + medication group, (6.09 +/- 3.78) scores in the western medicine group and (9.24 +/- 3.49) in the Chinese medicine group, the differences were significant in the Chinese medicine group compared with the western medicine group and the acupuncture + medication group (both P < 0.05). The differences in the scores after treatment were not significant statistically between the acupuncture + medication group and the western medicine group (both P > 0.05), but the cured and markedly effective rate in the acupuncture + medication group was superior to that in the western medicine group (P < 0.05). CONCLUSION: The alliance of acupuncture and medication achieves the definite efficacy on depression, characterized as less adverse reactions and better safety.

[Early clinical features of severe peripheral facial paralysis and acupuncture strategies].

In order to have a good grasp of rules of acupuncture for severe peripheral facial paralysis, the early clinical features of severe peripheral facial paralysis (Bell's palsy) are studied and analyzed from the aspect of injury level, injury degrees, clinical syndromes and symptoms; consequently, the treatment strategies with acupuncture are proposed. The severe peripheral facial paralysis is an important research area in clinic trials which verifies the effectiveness of acupuncture treatment.

[Clinical observation on the regularity of acupuncture-induced body-reduction in excess-heat-type obesity patients ].

OBJECTIVE: To investigate the regularity of acupuncture of the Stomach Meridian for excess-heat-type obesity. METHODS: Fifty-one patients with gastrointestine excess-heat syndrome were randomly divided into treatment group (n = 25) and control group (n = 26) according to the random number table method. Patients of treatment group were treated mainly with acupuncture of acupoints of the Stomach Meridian as bilateral Liangmen (ST 21), Huaroumen (ST 24), Tianshu (ST 25), Wailing (ST 26), etc. and those of control group were treated with acupuncture of Back-Shu and Front-Mu points as bilateral Weishu (BL 21), Dachangshu (BL 25), Xiaochangshu (BL 27), Hegu (LI 4), etc. which were also stimulated electrically with parameters of 40-100 Hz, 3- 10 mA and stimulation duration of 30 mm. The treatment was given once every other day continuously for 3 months. Body weight, body mass index (BMI), waistline and clinical symptoms were recorded before and after each course of the treatment. RESULTS: After the treatment, of the 25 and 26 cases in treatment and control groups, 5 and 0 were cured, 9 and 7 had marked improvement, 7 and 10 improved, 4 and 9 failed, with the effective rates being 84.00% and 65.38% separately. Compared with control group and pre-treatment of the same group, the waistline, body weight and BMI of treatment group decreased significantly (P < 0.05, 0.01). CONCLUSION: Acupuncture of acupoints of the Stomach Meridian has a good therapeutic effect in reducing body weight in gastrointestine excess-heat-type obesity patients.

[Some problems in the literature of acupuncture treatment of peripheral facial paralysis and suggestions].

OBJECTIVE: To raise some problems in the literature of acupuncture and moxibustion for treatment of peripheral facial paralysis and improving suggestions. METHODS: The CHKD system was used to search out 817 papers of acupuncture for treatment of peripheral facial paralysist, which were reviewed, organized and summarized. RESULTS: In the literature of acupuncture for treatment of peripheral facial paralysis, insufficient understanding in the relative information of clinical data, naming of intractable facial paralysis, differentiation between of the sequelae and complications, the criteria or cproblems in the literature and so on were found. CONCLUSION: There are some common and representative problems in these papers of acupuncture for treatment of peripheral facial paralysis, which may result in incorrect opinion about the theory and clinical study of acupuncture for treatment of peripheral facial paralysis. Sufficiently considering and resolving the above problems can not only improve the quality of the papers on acupuncture for treatment of peripheral facial paralysis as a whole, but also have active influence on both treatment and diagnosis of this disease.

[Programme of criteria for clinical evaluation and assessment of therapeutic effects of peripheral facial paralysis].

OBJECTIVE: To establish perfect criteria for clinical evaluation and assessment of therapeutic effects of peripheral facial paralysis. METHODS: With reference to relative evaluating criteria, and in combination with self-characteristics of TCM and functional characteristics of facial nerves, and via clinical repeated investigation and control evaluation, put forward a scale for peripheral facial nerve rating, criteria for facial nerve grading, criteria of syndrome types and criteria of assessment of therapeutic effects. RESULTS: Clinical preliminary determination indicated that the assessment contents were characterized by rational reference, convenience and objectiveness. CONCLUSION: The program can used as criteria for clinical evaluation and assessment of therapeutic effects, but it needs support of clinical effectiveness and determination of confidence, and multi-central studies.