Objective:To investigate the function of miR-5581-5p and its interaction with tripartite motif 22 (TRIM22) during the terminal differentiation of human acute promyelocytic leukemia (APL) cells into granulocytes.Methods:APL cells (NB4) were differentiated into granulocytes by all-trans retinoic acid (ATRA), using dimethylsulfoxide (DMSO) as the control. The expression of TRIM22 was detected by real-time fluorescent quantitative PCR (qRT-PCR) and Western blotting, and the expression of miR-5581-5p was detected by qRT-PCR during cell differentiation. miRNA expression was regulated by cell transfection with miR-5581-5p mimic and inhibitor, and negative control was set, and qRT-PCR was used to verify the regulatory effect. Luciferase binding assay was performed to detect the presence of targeted binding. Western blotting was used to detect the expression of TRIM22 after miRNA differential expression. Flow cytometry was used to detect the effects of the regulation of miR-5581-5p on the differentiation of NB4 cells induced by ATRA.Results:After ATRA induced NB4 cells to differentiate into granulocytes, the gene expression level of TRIM22 was significantly higher than that of the control group (24.56±2.80 vs. 1.02±0.13; t=8.392, P=0.001). The level of protein expression was also significantly higher than that of the control group (0.80±0.01 vs. 0.17±0.01; t=44.900, P<0.001). The expression level of miR-5581-5p in NB4 cells differentiation group was significantly lower than that in the control group (0.14±0.02 vs. 1.01±0.08; t=10.840, P<0.001). The results of the dual luciferase reporter gene showed that the luciferase activity of the co-transfected miR-5581-5p mimic and TRIM22 WT group was significantly lower than that of the co-transfected miR-5581-5p mimic and TRIM22 MUT group (0.73±0.02 vs. 0.98±0.03; t=7.534, P=0.002). Western blotting showed that after transfection with miR-5581-5p inhibitor, the expression of TRIM22 was significantly higher than that of the negative control (0.44±0.01 vs. 0.21±0.01; t=18.290, P<0.001). While after transfection with miR-5581-5p mimic, the expression of TRIM22 decreased significantly compared with the negative control (0.62±0.01 vs. 0.80±0.02; t=6.402, P=0.003). CD11b expression of miR-5581-5p mimic group after ATRA treatment was significantly lower than that of the control group (45.80±1.80 vs. 56.61±1.88; t=4.159, P=0.014). The expression of CD11b in miR-5581-5p inhibitor group was significantly higher than that in the control group (66.48±2.54 vs. 52.60±1.70; t=4.539, P=0.011). Conclusion:miRNA-5581-5p can bind to TRIM22 3′UTR and negatively regulate TRIM22 expression. The decrease of miR-5581-5p can increase the expression of TRIM22, then promote the differentiation of ATRA-induced NB4 cells into granulocytes.

Objective:To observe the changes of insulin secretion in the early stage of severe scald in rats, and to explore its signal transduction mechanism.Methods:Twenty-four male Wistar rats aged 7 weeks were divided into sham injury alone (SIA) group, sham injury+ BPV (HOpic) (SIB) group, scald alone (SA) group, and scald+ BPV (HOpic) (SB) group using the random number table, with 6 rats in each group. Full-thickness scald of 50% total body surface area was inflicted in rats of SA and SB groups by a 6-s immersion of the abdomen and a 12-s immersion of the back in 94 ℃ hot water. Rats in SIA and SIB groups received sham injuries through immersion of the back and abdomen in 37 ℃ warm water for 6 and 12 seconds respectively. From 0 (immediately) to 2 day (s) after injury, the rats in groups SB and SIB were intraperitoneally injected with the phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway enhancer BPV (HOpic) solution (0.5 mg/mL) at the dosage of 0.6 mg/kg once a day, and the rats in groups SA and SIA were intraperitoneally injected with the same volume of dimethyl sulfoxide once a day. At post injury hour (PIH) 72, the tail blood of rats was sampled for measuring fasting blood glucose (FBG) with a glucometer, and the pancreatic tissue samples of rats was harvested for observing the pathological manifestations of islets by hematoxylin-eosin staining, counting the docked granules per 10 μm membrane of islet beta cells and calculating the proportion of insulin vesicles through the observation of the ultrastructure of islet beta cells by transmission electron microscope, and detecting the phosphorylation level of Akt in the pancreatic PI3K/Akt signaling pathway by Western blotting. Data were statistically analyzed with one-way analysis of variance and least significant difference test.Results:(1) At PIH 72, the rat FBG levels in SIA and SIB groups were normal and similar ( P>0.05). Compared with the levels of those two groups, the rat FBG level in SA group was increased significantly ( P<0.01), while the level in SB group showed no obvious change ( P>0.05). Compared with that in SA group, the rat FBG level in SB group was decreased significantly ( P<0.01). (2) At PIH 72, the morphology of rat islets was complete and the islet cells distributed regularly in SIA and SIB groups. Compared with those in SIA and SIB groups, the morphology of rat islets was incomplete, the insulin vesicles in islets were common, the islet cells distributed irregularly, and the cytoplasm of some islet beta cells was lightly stained or translucent in SA group; the morphology of islets in SB group did not change obviously. Compared with those in SA group, the morphology of islets was comparatively complete, the insulin vesicles in islets were less common, the islet cells distributed comparatively regularly, and the lightly stained or translucent cytoplasm of islet beta cells was less in SB group. (3) At PIH 72, the number of docked granules per 10 μm membrane of rat islet beta cells and the proportion of insulin vesicles in SIA and SIB groups were similar ( P>0.05). Compared with those in SIA and SIB groups, the number of docked granules per 10 μm membrane of rat islet beta cells in SA group was decreased significantly ( P<0.01), while the proportion of insulin vesicles was increased significantly ( P<0.01); the number of docked granules per 10 μm membrane of rat islet beta cells in SB group was obviously decreased ( P<0.05), while the proportion of insulin vesicles did not change obviously ( P>0.05). Compared with those in SA group, the number of docked granules per 10 μm membrane of rat islet beta cells in SB group was significantly increased ( P<0.01), while the proportion of insulin vesicles was significantly decreased ( P<0.01). (4) At PIH 72, the phosphorylation levels of Akt in SIA, SIB, SA, and SB groups were 0.91±0.03, 0.98±0.03, 0.78±0.08, and 0.87±0.08, respectively. Compared with that in SIA group, the phosphorylation level of Akt was increased obviously in SIB group ( P<0.05) but was decreased significantly in SA group ( P<0.01), while the level in SB group did not change obviously ( P>0.05). Compared with the level in SIB group, the phosphorylation levels of Akt in SA and SB groups were decreased significantly ( P<0.01). Compared with that in SA group, the phosphorylation level of Akt in SB group was increased significantly ( P<0.05). Conclusions:At the early stage post severe scald in rats, the activity of the pancreatic PI3K/Akt signaling pathway and the function of insulin secretion are reduced. Improving the activity of the pancreatic PI3K/Akt signaling pathway in rats can ameliorate the function of insulin secretion and recover the physiological level of blood glucose.

Objective To explore the effect and potential mechanisms of melatonin combined with gemcitabine on the chemosensitivity of human pancreatic cancer cell line PANC-1.Methods Human pancreatic cancer cell line PANC-1 was trea-ted with gemcitabine alone or in combination with melatonin.Cell viability was assessed using CCK-8.Effect of melatonin and gem-citabine alone or in combination on the clonogenic capacity of PANC-1 cells were observed through colony formation experiments.Scratch assays and transwell experiments were conducted to evaluate cell migration ability.Reactive oxygen species(ROS)and mitochondrial membrane point JC-1 assay kit were used to determine reactive oxygen species synthesis and membrane potential levels.Intracellular Fe2+level was measured using ferrous ion fluorescent probe.The protein expression levels of LC3,P62,GPX4 and SLC7A11 in different treatment groups were detected by immunofluorescence and Western blotting.Results CCK-8 results showed that the viability of PANC-1 cells was inhibited by gemcitabine alone after 48 h and 72 h of treatment in a time-and dose-dependent manner.The cell viability of gemcitabine combined with melatonin group was significantly lower than that of gemcitabine group,and the cell viability decreased with the increase of melatonin concentration.Scratch assays,transwell experiments,and plate colony formation assay results demonstrated that the proliferation and migration of cells in the gemcitabine combined with the me-latonin group were significantly inhibited compared with the gemcitabine group.The levels of reactive oxygen species and Fe2+in PANC-1 in gemcitabine combined with the melatonin group were higher than those in the gemcitabine group,and the mitochondri-al membrane potential was significantly decreased(P＜0.01).Western blotting and immunofluorescence results showed that the ra-tio of autophagy-related protein LC3-Ⅱ/LC3-Ⅰ in gemcitabine combined with the melatonin group was lower than that in the gem-citabine group,and the expression of P62 was up-regulated,and the expression of anti-iron death-related protein GPX4 and SLC7A11 was significantly inhibited(P＜0.05),suggesting that melatonin combined with gemcitabine can inhibit autophagy and promote ferroptosis in PANC-1 cells.Conclusion Melatonin enhances the chemosensitivity of pancreatic cancer cell PANC-1 to gemcitabine by inhibiting autophagy and promoting ferroptosis of tumor cells.

Objective To qualitatively analyze the main chemical components in compound Jinqiancao granules by ultra high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS). Methods XBridge BEH C₁₈ column (2.1 mm×100 mm, 2.5 µm) was used for chromatographic separation. The mobile phase was composed of 0.1% formic acid water and 0.1% formic acid-acetonitrile, gradient elution, and the flow rate was 0.4 ml/min. Mass spectrometry was characterized by Quadrupole time-of-flight mass spectrometry (Q-TOF/MS) and positive ion mode scanning. Results Under the optimized LC/MS condition, 47 components in compound Jinqiancao granules were identified. The isomers were distinguished by software calculation. The source of medicinal materials was assigned. Conclusion A rapid and efficient analytical method was established for the identification of chemical components in compound Jinqiancao granules by UHPLC-Q-TOF/MS.

ObjectiveTo evaluate the clinical efficacy and safety of traditional Chinese medicine （TCM） suppository combined with Yishen Tongluo Qingkang decoction in the treatment of immune infertility. MethodA total of 116 patients meeting the inclusion criteria of this study were randomly divided into an observation group （58 cases） and a control group （58 cases）. The observation group was treated with TCM suppository combined with Yishen Tongluo Qingkang decoction，and the control group was treated with prednisone acetate tablets. Both groups were treated for 12 weeks and followed up six months after treatment. Semen samples of the patients were collected before and after treatment，and the pregnancy status of their spouses，negative conversion rate of seminal plasma anti-sperm antibody （AsAb），sperm concentration，motility，percentage of forward motile sperm，sperm acrosin activity， and incidence of adverse reactions were compared between the two groups. ResultA total of 104 patients completed the study，including 53 cases in the observation group and 51 cases in the control group. Before treatment，the baseline data of the two groups were balanced. After treatment，the total effective rate of the observation group was 92.45%，which was higher than that of the control group （76.47%）（P<0.05），and the negative conversion rate of AsAb in the observation group was higher than that in the control group，but the difference was not statistically significant. After treatment，the sperm motility， percentage of forward motile sperm， and sperm acrosin activity increased in the two groups（P<0.05），and the sperm concentration in the observation group increased （P<0.05）. There was no significant difference in sperm concentration in the control group. After treatment，the sperm concentration，motility，percentage of forward motile sperm， and acrosin activity in the observation group were better than those in the control group （P<0.05）. During the trial，the incidence of adverse reactions in the observation group was lower than that in the control group （P<0.05）. ConclusionTCM suppository combined with Yishen Tongluo Qingkang decoction can significantly increase the negative conversion rate of AsAb and improve the quality of semen in patients with immune infertility.

ObjectiveTo evaluate the clinical effects of Yishen Tongluo Prescription in treating patients with idiopathic asthenozoospermia and teratozoospermia due to kidney deficiency and collateral obstruction， as well as the effects of this formula on biochemical indices and oxidative stress of seminal plasma. MethodsPatients who fulfilled the inclusion criteria were randomly allocated by the random number table method into an observation group （61 patients， including 2 dropped out and 59 completed） and a control group （61 patients， including 2 dropped out， 1 excluded， and 58 completed）. The observation group took Yishen Tongluo Prescription orally， 1 dose/day， and the control group took levocarnitine oral solution， 10 mL/time， 3 times/day. Both groups received a treatment course of 12 weeks and were followed up for 6 weeks. The percentage of progressive motile sperms （PR）， percentage of sperms with normal morphology， seminal plasma oxidative stress indicators ［malondialdehyde （MDA） and superoxide dismutase （SOD）］， seminal plasma biochemical indicators ［acid phosphatase （ACP）， zinc （Zn）， fructose （Fru）， and alpha-glucosidase （α-Glu）］， and traditional Chinese medicine （TCM） symptom scores were recorded for both groups before and after treatment. ResultsThe total response rate in the observation group was 76.3% （45/59）， which was higher than that （44.8%， 26/58） in the control group （χ²=12.120， P<0.05）. After treatment， both groups of patients showed increases in PR， percentage of sperms with normal morphology， and seminal plasma SOD level （P<0.05）， which were higher in the observation group than in the control group （P<0.05）. The MDA level and TCM symptom scores decreased in both groups after treatment （P<0.05）， and the decreases were more obvious in the observation group than in the control group （P<0.05）. After treatment， the seminal plasma levels of ACP， Zn， Fru， and α-Glu in the observation group （P<0.05） and that of α-Glu in the control group （P<0.05） rose. Moreover， the observation group had higher seminal plasma levels of ACP， Zn， Fru， and α-Glu than the control group （P<0.05）. Neither group showed obvious adverse reactions. ConclusionIn summaryYishen Tongluo prescription is able to improve the sperm vitality and reduce the sperm abnormality rate in infertile patients with idiopathic asthenozoospermia and teratozoospermia due to kidney deficiency and collateral obstruction. It may exert therapeutic effects by regulating the microenvironments of the epididymis， prostate， and seminal vesicles， thereby recovering the seminal plasma biochemical levels， enhancing the antioxidant capacity of seminal plasma， clearing excessive reactive oxygen species， and alleviating oxidative stress.

ObjectiveTo observe the clinical efficacy of herb-partitioned moxibustion on the navel combined with Yishen Tongluo prescription in the treatment of infertility induced by idiopathic asthenozoospermia （iAZS） with kidney-Yang deficiency and collateral obstruction syndrome and its effect on sperm DNA damage and superoxide dismutase （SOD） in the seminal plasma. MethodsA total of 112 eligible patients who met the inclusion criteria were randomly divided into an observation group （56 cases） and a control group （56 cases）. The patients in the observation group were treated with herb-partitioned moxibustion on the navel combined with Yishen Tongluo prescription，while those in the control group received levocarnitine oral liquid. The primary observation indicators included spouse pregnancy rate，progressive motility （PR），and total sperm motility，and the secondary observation indicators included sperm DNA fragmentation index （DFI），SOD in the seminal plasma， and improvement of TCM syndromes. The treatment cycle was 12 weeks. Before and after treatment，the PR，total sperm motility，sperm DFI，SOD in the seminal plasma， and TCM syndrome scores were recorded. The patients were followed up for 12 weeks and the pregnancy status of spouses within 24 weeks （half a year） was recorded. The clinical efficacy of the two groups was evaluated. ResultThe pregnancy rate of spouses in the observation group was 15.69% （8/51）， higher than 3.85% （2/52） in the control group （χ²=4.118，P<0.05）. The total effective rate of the observation group was 88.24%（45/51）， superior to 69.23% （36/52）in the control group （Z=-3.402，P<0.01）. After treatment， PR， total sperm motility，sperm DFI， SOD in the seminal plasma， and TCM syndromes of the two groups were improved compared with those before treatment （P<0.05）， and the observation group was superior to the control group （P<0.05）. ConclusionHerb-partitioned moxibustion on the navel combined with Yishen Tongluo prescription in the treatment of iAZS-induced infertility patients with kidney-Yang deficiency and collateral obstruction syndrome can increase PR，total sperm motility， and SOD level in the seminal plasma， reduce sperm DFI，improve the TCM symptoms of patients， and improve the pregnancy rate of spouses. The mechanism may be attributed to the fact that this treatment can increase the SOD level in the seminal plasma of patients，enhance the body's antioxidant function，protect sperm from oxidative stress damage，and reduce sperm DFI.

Helicobacter pylori (H. pylori), creating a global infection rate over 50%, presents great challenges in clinical therapies due to its complex pathological microenvironment in vivo. To improve the eradication efficacy, herein we fabricated a pharmaceutical vesicle RHL/Cl-Ch-cal where cholesterol-PEG, calcitriol and first-line antibiotic clarithromycin were co-loaded in the rhamnolipid-composed outer lipid layer. RHL/Cl-Ch-cal could quickly penetrate through gastric mucus layer to reach H. pylori infection sites, and then effectively destroyed the architecture of H. pylori biofilms, killed dispersed H. pylori and inhibited the re-adhesion of residual bacteria (called biofilms eradication tetralogy). Moreover, RHL/Cl-Ch-cal activated the host immune response to H. pylori by replenishing cholesterol to repair lipid raft on the cell membrane of host epithelial cells. Finally, RHL/Cl-Ch-cal killed the intracellular H. pylori through recovering the lysosomal acidification and assisting degradation. In experiments, RHL/Cl-Ch-cal demonstrated prominent anti-H. pylori efficacy in the classical H. pylori-infected mice model. Therefore, the study provides a "comprehensive attack" strategy for anti-H. pylori therapies including biofilms eradication tetralogy, immune activation and intracellular bacteria killing.

ObjectiveTo observe the clinical effect of governor meridian moxibustion combined with modified Youguiwan in treating erectile dysfunction of kidney Yang deficiency type and its influence on the level of sex hormones and penile hemodynamics. MethodA total of 120 patients were randomized into the control group （60 cases in total， 3 dropouts/lost to follow-ups， 57 finally included） and treatment group （60 cases in total， 2 dropouts/lost to follow-ups， 58 finally included） with the random number table method. Both groups received governor meridian moxibustion （1 time/w， a total of 4 times）. In addition， the control group took oral compound Xuanju capsule （3 capsules/time， 3 times/d）， and the treatment group was given Jiawei Youguiwan （Chinese medicine decoction， 1 dose/d）. The administration lasted 4 weeks for both groups. The scores of 5^th edition international index of erectile function （IIEF-5）， erection quality scale （EQS）， and erectile hardness score （EHS）， traditional Chinese medicine （TCM） syndrome score， levels of sex hormones ［testosterone （T）， follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， prolactin （PRL）］， and parameters of penile hemodynamics ［peak systolic velocity （PSV）， end-diastolic volume （EDV）， and resistance index （RI）］ were recorded. ResultAfter treatment， the total effective rate was 87.93% （51/58） in the treatment group， higher than the 71.93% （41/57） in the control group （χ²=4.600 3， P<0.05）. After treatment， compared with before treatment，the treatment group registered increase in the scores of IIEF-5， EQS， and EHS （P<0.01）， decrease in TCM syndrome score （P<0.01）， rise of serum T level （P<0.05）， reduction in the levels of FSH， LH， and PRL （P<0.05）， increase in the levels of PSV and RI （P<0.05）， and down-regulation of EDV level （P<0.05）. ConclusionGovernor meridian moxibustion combined with modified Youguiwan can effectively improve the penile erectile function， erectile hardness， and erection quality， and relieve the clinical symptoms of erectile dysfunction patients of kidney Yang deficiency type.