Objective To investigate the effects of metformin(Met)on the proliferation of pancreatic cancer cells under different glucose concentration culture conditions,and to find the potential role of miR-139-5p in the process.Methods PANC-1 cells were treated with different concentrations of metformin(0/5/10/20 mmol/L)in 25 mmol/L(high-glucose group,HG)or 5 mmol/L(normal-glucose group,NG)glucose culture,cell proliferation,apoptosis,migration and cell cycle were detected after 48 h.The expression of miR-139-5p was quantitatively detected by RT-qPCR,and the miR-139-5p mimics were transfected into PANC-1 cells to clarify the role of miR-139-5p.Results Metformin inhibited the proliferation,promoted apoptosis,and induced S phase and G2/M phase arrest of PANC-1 cells under in high glucose and normal glucose culture conditions,and its anti-proliferation and pro-apoptosis effects were more significant in the normal glucose groups.The expression of miR-139-5p was up-regu-lated by metformin treatment in normal but not in high glucose culture.Further studies showed that miR-139-5p mimics inhibited of PANC-1 cells proliferation without metformin pre-incubation and enhanced the anti-prolifera-tion effect of 5 mmol/L metformin.The pro-apoptotic effect of 10 mmol/L metformin in normal glucose culture conditions.Conclusions In normal-glucose culture conditions,metformin can inhibit proliferation,induce apop-tosis and cell cycle arrest of PANC-1 cells more significantly than in higher-glucose culture,which may be partly related to the up-regulation of miR-139-5p.

Objective To identify the relationship between physical activity,insulin resistance and cardiovascular risk in individuals with different glucose tolerance status and to provide evidence for exercise intervention in people with different glucose tolerance status.Methods A total of 691 patients with different glucose metabolism status were recruited as subjects of the research.Spearman correlation analysis was used to study the relationship between exercise frequency and insulin resistance,insulin sensitivity,neck circumference(NC)and neck circumference height ratio(NHtR)in the subjects with different glucose metabolism status,the relationship between NC and insu-lin resistance and insulin sensitivity in different glucose metabolism groups.Results 171(62.9%)Subjects with diabetes were intervened by exercised every day.Spearman correlation analysis showed the correlation between exer-cise frequency and tri-glyceride triglyceride-glucose index(TyG index)(r=-0.120,P＜0.05)and NC(r=-0.168,P＜0.05)were negatively correlated.In subjects with diabetes,NC was positively correlated with triglyc-erides(TG)(r=-0.100,P＜0.05),homeostasis model assessment of insulin resistance(HOMA-R)(r=-0.163,P＜0.05),total cholesterol/high-density lipoprotein(TC/HDL-C)(r=-0.214,P＜0.05)and TyG index(r=-0.156,P＜0.05).Conclusions Increased frequency of exercise is associated with reduced NC,improved insulin resistance,and cardiovascular risk factors in subjects of our team with newly diagnosed diabetes.Exercise has no significant effect on insulin resistance of subjects with normal glucose tolerance and pre-diabetes.

Objective To investigate the clinical effects and complications of endoscopic tandem ligation and hemorrhoids ligation.Methods Clinical data of 120 patients with grade Ⅰ～Ⅲ internal hemorrhoids who were hospitalized due to hematochezia from November 2021 to March 2023 were retrospectively analyzed.According to the different surgical methods,60 patients who received endoscopic hemorrhoid ligation were divided into control group;60 patients who underwent endoscopic tandem ligation were divided into treatment group.The effective rate of hemorrhoidal hemorrhage,the effect of internal hemorrhoidal prolapse and the incidence of postoperative complications were compared between two groups within 6 months after operation.Results The effective rate of internal hemorrhoidal bleeding was 100.00％in both groups,and the efficacy of the treatment group in the treatment of degree Ⅱ internal hemorrhoidal prolapse was similar to that in the control group(P＞0.05),but the treatment group was significantly better than the control group in the treatment of degree Ⅲ internal hemorrhoidal prolapse(P＜0.01),the postoperative anal pain in the treatment group was significantly more than the control group(P＜0.05);There were no significant differences in postoperative perianal pain,urinary retention,and abnormal defecation between the two groups(P＞0.05).No thrombotic external hemorrhoids occured in two groups.Conclusion The short-term efficacy of endoscopic tandem ligation and hemorrhoidal ligation in the treatment of internal hemorrhoidal bleeding is similar and both achieved satisfactory results,the short-term efficacy of the two in the treatment of degree Ⅱ internal hemorrhoidal prolapse is similar,the short-term efficacy of the former in the treatment of degree Ⅲ internal hemorrhoidal prolapse is significantly better,the feeling of anal heaviness in the treatment group is more than that in the control group,and there is no striking difference in the other complications.

Objective:Both type 1 diabetes and type 2 diabetes are associated with abnormal bone metabolism, but they have different pathogenic mechanisms. Sclerostin(SOST), Dickkopf-related protein 1(DKK-1), and irisin are newly discovered factors involved in the regulation of bone metabolism. This study aims to compare the differences in serum levels of SOST, DKK-1, and irisin between patients with type 1 diabetes and type 2 diabetes.Methods:This cross-sectional study included 101 patients with type 1 diabetes who visited the Endocrinology Department of Peking Union Medical College Hospital from 2017 to 2019, as well as 55 patients with type 2 diabetes and 59 individuals with normal glucose tolerance who were confirmed through an oral glucose tolerance test as part of the Beijing Changping Community Type 2 Diabetes Management Program from 2014 to 2015. Enzyme-linked immunosorbent assay(ELISA) was used to measure the levels of SOST, DKK-1, and irisin.Results:There were more female participants than male participants, with an average age of 49 years. The group with type 1 diabetes had a longer duration of illness( P<0.001) and higher HbA 1C levels( P<0.001) compared to the group with type 2 diabetes, and there was no statistical difference in age between the two groups. Both the type 1 diabetes and type 2 diabetes groups had lower levels of serum procollagen type 1 N-terminal propeptide(P1NP) compared to the control group [(8 579±400)pg/mL, (7 268±552)pg/mL vs(10 051±618)pg/mL, P=0.039, P=0.001]; But the β isomer of C-terminal cross-linking telopeptide of type 1 collagen(β-CTX) showed no statistical difference compared to the control group. Patients with type 1 diabetes and type 2 diabetes had higher SOST than controls [(129.7±6.8)pg/mL, (104.8±6.8)pg/mL vs(85.9±5.3)pg/mL, P<0.001, P=0.030], the differences between the type 1 diabetes group and the control group lost statistical significance after adjusting for factors such as fasting blood glucose and lipid levels. There was no significant difference in SOST between type 1 diabetes and type 2 diabetes groups. There was no significant difference in DKK-1 among three groups, but DKK-1 in type 1 diabetes group was lower or tended to be lower than that in type 2 diabetes group. Serum irisin in patients with type 1 diabetes was higher than that in controls and patients with type 2 diabetes[(16.6±0.7)ng/mL vs (9.6±0.6)ng/mL, (9.8±0.6)ng/mL, both P<0.001], but there was no significant difference in irisin level between type 2 diabetes and controls. Conclusions:Patients with both type 1 and type 2 diabetes showed inhibition of the bone formation marker P1NP, while the bone resorption marker β-CTX did not significantly change. SOST levels were elevated or showed an increasing trend in both type 1 and type 2 diabetes patients, which may be related to the inhibition of bone formation. Additionally, type 1 diabetes patients had increased levels of irisin, which may be involved in abnormal bone turnover.

Objective:To investigate the efficacy of endoscopic histoacryl injection in cirrhotic patients with newly-developed esophagogastric varices (EGV) who have previously undergone splenectomy combined with pericardial devascularization.Methods:From January 2015 to January 2020, 125 cirrhotic patients with EGV treated with endoscopic histoacryl injection at the Department of Gastroenterology, Jinling Hospital, Medical School of Nanjing University, were included in the retrospective analysis. There were 45 patients in the group of splenectomy combined with pericardial devascularization (splenectomy group for short) and 80 patients in the non-splenectomy group. The efficacy of endoscopic treatment, postoperative variceal improvement, rebleeding rate, and complications were analyzed between the two groups.Results:Endoscopic histoacryl injection was successfully completed in all 125 patients, and the median volume of histoacryl was 4.5 mL. The overall effective rate in splenectomy and non-splenectomy group was 80.0% (36/45) and 57.5% (46/80), respectively. The difference in the number of significantly effective, effective, and ineffective cases between the two groups was statistically significant (16, 20, 9 cases, and 20, 26, 34 cases, respectively, χ 2=6.469, P=0.039). Two and 14 patients developed rebleeding in the splenectomy group and non-splenectomy group, respectively; and the difference in the rebleeding rate between the two groups was statistically significant (4.4% VS 17.5%, Log-rank P=0.039). No patient died within 1 year in either group, and no serious complications such as ectopic embolism occurred. Conclusion:After splenectomy combined with pericardial devascularization in cirrhotic patients with EGV and hypersplenism, the application of histoacryl has better short-term efficacy and can significantly reduce the rebleeding rate compared with the non-splenectomy group.

  Objective  By summarizing the clinical characteristics and follow-up outcomes of 5 patients with immune checkpoint inhibitor induced diabetes mellitus (ICI-DM) and reviewing the relevant literatures, the article aims at providing reference to clinicians in the diagnosis and treatment of the ICI-DM.  Methods  Clinical data of 5 patients with ICI-DM who were admitted to Peking Union Medical College Hospital from December 2018 to February 2023 and did retrospectively analyzed.  Results  Five patients with a mean age of (65±7)years received treatment by the programmed cell death 1 (PD-1) or its ligand inhibitor (PD-L1). The median time from the first immunotherapy to the discovery of elevated plasma glucose was 100 (43, 210)days, and the median cycle of immunotherapy was 7 (2.5, 10.5). The onset of the illness of all the 5 patients started with diabetic ketosis or ketoacidosis. At the onset, urine ketone bodies were positive, random plasma glucose was (36.36±15.89)mmol/L, glycosylated hemoglobin A1c (HbA1c)was (8.6%±0.66%), arterial blood pH was (7.28±0.16), and the median fasting C-peptide level was 0.09 (0.05, 0.32)μg/L. Five patients had an onset plasma glucose level of grade 3 or 4.Then, ICI treatment was discontinued in all patients and insulin therapy started. The daily dosage of insulin was (56±20)IU, supplemented with hypoglycemic drugs. After treatment, urine ketone body turned negative, pH value increased to normal range, and random plasma glucose decreased significantly (the median difference of random blood glucose before and after treatment was 21.30 mmol/L, P=0.043) showing that the treatment was effective. During the follow-up, all patients continued to use insulin. The PD-1 or PD-L1 inhibitors were restarted after hyperglycemia remission. The tumor condition was under control.  Conclusions  ICI-DM mainly occurs in patients who receive treatment with PD-1 or PD-L1 inhibitors usually with acute hyperglycemia whose laboratory tests indicate insulin secretion defects. Some patients had positive islet cell antibodies, glutamic acid decarboxylase antibodies and autoantibodies.Patients with positive autoantibodies needed early diagnosis and continuous insulin treatment. ICI treatment can be restarted after endocrinologists brought the blood glucose under control.

Objective:To investigate the effect of autophagy on the Treg/Th17 cell imbalance in mice with acute lung injury (ALI).Methods:Twenty-four male SD mice were randomly divided into the sham operation group (S group), sepsis group (Sep group) and autophagy inhibitor 3-methyladenine group (Sep +3-MA group). ALI model was prepared by LPS tracheal dripping method. The mouse pathological injury score mice were evaluated under light microscopy and the W/D ratio was calculated. The counts of Th17 cells and Treg cells in tracheoalveolar lavage fluid (BALF) of mice and the levels of related cytokines were detected by flow cytometry. The expressions of LC3-Ⅱ, Beclin-1 and p62 in Th17 cells and Treg cells in BALF were determined by Western blot.Results:CCompared with the S group, the lung histopathological score and W/D ratio of the Sep group and Sep+3-MA group increased ( P<0.05). Compared with the Sep group, the count of Th17 cells in BALF of the Sep +3-MA group decreased, while the count of Treg cells increased significantly with the progression of sepsis( P<0.05), and the levels of IL-17, IL-10 and TNF-α were significantly decreased ( P<0.05). TGF-β1 levels increased in the early stages of sepsis, but decreased significantly with the progression of sepsis( P<0.05). Compared with the Sep group, LC3-Ⅱ expression in BALF Th17 cells and Treg cells of the Sep+3-MA group showed a downward trend, but there was no statistical difference, while Beclin-1 expression significantly decreased ( P<0.05), and the expression of p62 significantly increased ( P<0.05). Conclusions:Abnormal activation of autophagy in Th17 cells and Treg cells is involved in the immune imbalance of Th17/Treg cells in ALI with sepsis. Inhibition of autophagy can restore the functions of Th17 cells and Treg cells, and improve the imbalance of Th17/Treg by inhibiting autophagy may become a new idea to control the pathogenesis and progression of immune disorders with sepsis.

Patients with end-stage liver disease (ESLD) are often accompanied by various complications such as sarcopenia and cachexia including lipopenia, and it was believed in the past that such status was associated with malnutrition, while recent studies have shown that myostatin (MSTN) is associated with the progression of ESLD. MSTN can lead to sarcopenia and cachexia by affecting the metabolism of glucose, fat, and protein and the number of myocytes, and it can be used as a screening indicator for hepatocellular carcinoma (HCC) and an indicator for disease progression. Intervention via the MSTN pathway might be an effective method for controlling sarcopenia and cachexia in patients with ESLD, and MSTN may be an effective indicator for predicting the progression of liver cirrhosis to HCC.

Objective:To evaluate the changes in CD4 + CD25 + regulatory T cells (Treg)/helper cell type 17 (Th17) during intestinal damage in septic rats. Methods:Forty-eight clean-grade Sprague-Dawley rats, aged 3 months, weighing 200-300 g, were divided into control group (group C)and sepsis group(group Sep), with 24 rats in each group.The modified cecal ligation and puncture method was used to establish the model of sepsis in anesthetized rats.Resuscitation was performed with normal saline after operation, and the right jugular vein catheterization was used for treatment of enteral nutrition in two groups.Six rats were sacrificed at 12, 24, 48 and 72 h after surgery (T 1-4), and mesenteric lymph nodes and colon were obtained.The percentage of Treg cells and Thl7 cells in mesenteric lymph nodes was detected by flow cytometry.The contents of interleukin-17 (IL-17), IL-23, transforming growth factor beta (TGF-β) and IL-10 in colon tissues were determined by enzyme-linked immunosorbent assay. Results:Compared with group C, the percentage of Treg cells was significantly decreased at T 1, the percentage of Treg cells at T 3, 4 and Th17 cells at T 2-4 were increased, the contents of IL-17 and IL-23 at T 1-4, TGF-β at T 3-4 and IL-10 at T 2-4 were increased, and TGF-β contents were decreased at T 2 in group Sep ( P<0.05). Conclusion:The mechanism of intestinal damage is related to Treg and Th17 cell imbalance in septic rats.

Objective To evaluate the effect of thymosin α1 (Tα1) on sepsis in rats.Methods A total of 120 pathogen-free healthy male Sprague-Dawley rats,weighing 250-300 g,aged 7 weeks,were divided into 3 groups using a random number table method:sham operation group (group S,n =24),sepsis group (group CLP,n=48) and Tα1 group (group T,n=48).Sepsis was induced by modified cecal ligation and puncture in anesthetized rats,and vena cava cannula was placed through the right jugular vein to connect the micropump infusion device.In group S,the cecum was only turned without ligation and perforation,and total venous nutrient solution 60 ml was intravenously infused daily for 3 days.After successful establishment of the model,total venous nutrient solution 60 ml was intravenously infused daily for 3 days in group CLP.After successful establishment of the model in group T,Tα1 0.18 mg/kg was subcutaneously injected daily at a fixed time,and total venous nutrient solution 60 ml was intravenously infused daily for 3 days.Twenty-four rats in group CLP and group T were selected,and the survival rate were observed within 3 days after establishing the model.At 12,24,36 and 72 h after establishing the model,6 rats were selected from each group,and blood samples were collected from the jugular vein for determination of the levels of regulatory T cells (Tregs) and T helper cell 17 (Th17) (using flow cytometry) and serum interleukin-10 (IL-10),IL-17 and tumor necrosis factor-alpha (TNF-α) concentrations (by enzymelinked immunosorbent assay),and the IL-10/TNF-cα ratio was calculated.Results Compared with group S,the level of Tregs in peripheral blood was significantly decreased at 24 h after establishing the model and was increased at 36 and 72 h after establishing the model,and the level of Th17 in peripheral blood and serum IL-10,IL-17 and TNF-α concentrations were increased at each time point after establishing the model,and the IL-10/TNF-α ratio was increased at 36 and 72 h after establishing the model in group CLP,and the levels of Tregs and Th17 in peripheral blood were significantly increased at 72 h after establishing the model,the concentrations of serum IL-10 and IL-17 were increased at each time point after establishing the model,serum TNF-α concentrations were increased at 12 and 24 h after establishing the model,and the IL-10/TNF-α ratio was increased at 36 and 72 h after establishing the model in group T (P＜0.05).Compared with group CLP,the levels of Tregs and Th17 in peripheral blood were significantly decreased,the serum IL-10 and IL-17 concentrations and IL-10/TNF-α ratio were decreased at 36 and 72 h after establishing the model,serum TNF-α concentrations were decreased at 72 h after establishing the model,and the survival rate was increased within 3 days after establishing the model in group T (P＜0.05).Conclusion Tα1 can reduce sepsis through improving the cellular immune function in rats.