Abstract: Objective To observe the effects of Qingre Runzao oral liquid on the numbers of type Ⅱ intrinsic lymphocytes (ILC2s) and their upstream and downstream cytokines in asthmatic mice sensitized by ovalbumin. Methods Female BALB/ c mice were divided into the blank control group, model control group, positive control group (dexamethasone group, 1 mg / (kg·d)) and low- and high-dose Qingre Runzao oral liquid groups (4. 32 and 8. 64 g / (kg ·d)). The asthma mouse model was induced using ovalbumin as a sensitizer, and pathological changes in lung tissue were observed by hematoxylin and eosin staining. Levels of the upstream and downstream cytokines of ILC2s Eotaxin, interleukin (IL)-25, IL-33, thymic stromal lymphopoietin ( TSLP), IL-4, IL-5 and IL-6 were detected in mouse bronchoalveolar lavage fluid ( BALF) using enzyme-linked immunosorbent assay. The expression of IL-33 and IL-5 in lung tissue was detected by immunohistochemical method . Flow cytometry and immunofluorescence staining were used to assess the number and production of ILC2s in lung tissue. Results Compared with the blank control group, the model control group showed pathological changes of inflammatory cell infiltration in the lung tissue and bronchial wall thickening. Furthermore, levels of the upstream and downstream cytokines Eotaxin, IL-25, IL-33, TSLP, IL-4, IL-5 and IL-6 were increased in BALF (P< 0. 01). The expression of IL-33 and IL-5 was increased in lung tissue (P<0. 01), and the number and production of ILC2s was also increased (P<0. 01). Compared with the model control group, both the low and high doses of Qingre Runzao oral liquid significantly improved the pathological injury to lung tissue in asthmatic mice, decreased the expression of upstream and downstream cytokines of ILC2s in BALF (P<0. 01), downregulated the expression of IL-33 and IL-5 in lung tissue (P<0. 05, P<0. 01), and decreased the number and production of ILC2s (P<0. 01). Conclusions Qingre Runzao oral liquid can reduce the numbers of ILC2s and their upstream and downstream cytokines in ovalbumin-induced asthmatic mice. It can also downregulate the expression of IL-33 and IL-5 in lung tissue, thus playing a role in the prevention and treatment of asthma.

Abstract: Objective To investigate the anti-atopic dermatitis (AD) effects of oxymatrine in a murine model of AD. Methods (1) Kunming mice were randomly divided into five groups: Blank Control group(BC), model control group (DEX), positive control group(DXM), OXY-L and OXY-H groups (50 and 100 mg / kg) oxymatrine groups. Vehicle or oxymatrine was orally administered twice daily for 7 times. Ten minutes after the last administration, 50 μL of dextran was injected subcutaneously into the left hind foot pad of each mouse. The number of times the mouse licked the hind foot within 15 min was observed and recorded. (2) BALB/ c mice were randomly divided into 6 groups: blank control group(BC), model control group(DNCB), positive control group (DXM), OXY-L, OXY-M and OXY-H groups ( 25, 50 and 100 mg / kg) oxymatrine groups. Except for the blank control group, the other groups were treated with 2,4-dinitrochlorobenzene (DNCB) solution repeatedly to establish the AD model. From the first day of the experiment, vehicle or drugs were orally administered once daily for 14 d. Changes in weight were observed and recorded, and the organ coefficients of the spleen and thymus were determined. The contents of total IgE and T helper 2 (Th2) cytokines in the peripheral blood were detected using enzyme- linked immunosorbent assay kits. In addition, hematoxylin and eosin staining was used to observe the pathological changes of skin lesions, and toluidine blue staining was used to evaluate the infiltration of mast cells in skin lesions. Results Oral administration of oxymatrine significantly decreased the number of scratches in the itching model mice (P<0. 01). After 14 d of treatment with oxymatrine by gavage, the eczema-like dermatitis of the AD model mice was relieved, and thickening of the dermis, infiltration of inflammatory and mast cells, and intercellular edema were reduced. The expression of total IgE and Th2-type cytokines, such as interleukin IL-4 and IL-13, was significantly lowered in the peripheral blood ( P< 0. 05). Conclusions Traditional Chinese medicine refers to the experience and knowledge summarized by Chinese people in the long-term practice of diagnosis and treatment of diseases. Oxymatrine, an effective component of the commonly used Chinese medicine Sophora flavescens, has a therapeutic effect on AD mice. Its mechanism is related to a reduction in the infiltration of mast cells in the skin dermis and the reduced expression of Th2-type cytokines.

Abstract: Objective To investigate the effects of Visfatin on glycolipid metabolism and insulin resistance in the skeletal muscle of diabetic KKAy mice, preliminarily analyze the possible mechanism of action through the PI3K/ Akt / FoxO1 pathway. Methods Eight-week-old male C57BL/ 6J mice were randomly divided into a normal diet (ND) group, a normal diet+Visfatin (ND+V) group, a high-fat (HFD) group, and a high-fat+Visfatin (HFD+V) group. When the model was generated, male KKAy mice of the same age were randomly divided into a diabetes mellitus (DM) group and diabetes mellitus + Visfatin (DM+V) group. Among them, the ND+V, HFD+V and DM+V groups were intraperitoneally injected with recombinant Visfatin protein for 3 consecutive days. Fasting blood glucose (FBG), postprandial blood glucose(PBG), total cholesterol (TC), triglyceride (TG), free fatty acids(FFA) and fasting insulin(FIns) were detected in each group of mice. The glucose tolerance test (GTT) and insulin tolerance test (ITT) were measured; meanwhile, the area under curve (AUC) and homeostasis model assessment for insulin resistance (HOMA-IR) were calculated. At the same time, the expression levels of phosphatidylinositol 3 kinase (PI3K), protein kinase B (Akt), forkhead box transcription factor 1 (FoxO1), pyruvate dehydrogenase kinase 4 (PDK4), phosphoenolpyruvate carboxykinase (PEPCK) and other related molecules were detected by RT-qPCR and Western blot. Results Compared with the ND group and the HFD group, the body weight, food intake, FBG, PBG, TC, TG, FFA, FIns, AUC_GTT , AUC_ITT and HOMA-IR levels in the DM group were significantly increased (P< 0. 05). Compared with DM group, FBG, TC, TG, AUC_GTT , AUC_ITT , and HOMA-IR levels in DM+V group were significantly reduced (P<0. 05). Compared with the ND group and the HFD group, the mRNA expression levels of P13K and Akt in the DM group were significantly decreased (P<0. 001), but the mRNA expression levels of FoxO1 were increased (P<0. 05). Compared with the DM group, P13K and Akt mRNA expression in the DM+V group was obviously increased, but the mRNA expression of FoxO1 was significantly reduced (P<0. 01). Compared with the ND group, PI3K_110α and p-Akt expression in the DM group was reduced, and the protein expression levels of FoxO1, p-FoxO1 and PDK4 were increased. Compared with the HFD group, p-Akt and p-FoxO1 expression was lower in the DM group, and the protein expression levels of FoxO1, PDK4 and PEPCK were increased (P<0. 05). While compared with the DM group, PI3K_110α , p-Akt and p-FoxO1 expression were increased in the DM+V group, but the protein expression levels of FoxO1, PDK4 and PEPCK were reduced (P<0. 05). Conclusions Visfatin may downregulate PDK4 expression in the skeletal muscle of diabetic mice through the PI3K/ Akt / FoxO1 pathway and has a role in improving glucolipid metabolism and insulin resistance.

Abstract: Objective The present study aimed to investigate the effects of “smoothen meridian and clear turbid dampness” (SMCTD) acupuncture on a model of acute gouty arthritis and explore its underlying mechanisms. Methods The model of acute gouty arthritis was induced in rats by monosodium urate (MSU) crystal injection. Next, the rats were treated with SMCTD acupuncture for 7 d or orally administered colchicine ( 1 mg / kg) for 7 d. Results MSU crystal injection induced a significant elevation in the concentrations of the serum proinflammatory cytokines interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α). Treatment with SMCTD acupuncture significantly reversed these MSU crystal- induced alterations (P<0. 05). The therapeutic effects of SMCTD acupuncture were further examined by histopathological examination. Exploration of the underlying mechanisms of its actions revealed that SMCTD acupuncture inhibited the activation of the nuclear factor κB(NF-κB) pathway, which was induced by MSU crystal injection. SMCTD acupuncture also significantly inhibited the activation of NF-κB p65 and the degradation of protein κB-α (IκBα) after the MSU crystal challenge. Conclusions Taken together, our findings demonstrate that SMCTD acupuncture has beneficial effects on MSU crystal-induced acute gouty arthritis, probably by inhibiting activation of the NF-κB/ IκBα signaling pathway.

Abstract: Objective To explore the effects of four different modeling method on endometrial receptivity, and establish a stable and effective experimental animal model of intrauterine adhesion ( IUA) with clinicopathological characteristics. Methods New Zealand white rabbits were randomly divided into 5 groups, according to different method of damage: chemical, infection, mechanical, thermal, and sham-operated ( control) groups. Rabbit bilateral uterine tissue was collected 4, 7, 14 and 28 d after treatment, and pathological changes were examined on both sides of the endometrium, using hematoxylin-eosin and Masson staining analysis. The thickness and number of endometrial glands and fibrosis area ratios were determined to evaluate the severity of adhesions. Pregnancy rates and the number of embryo implantations were used to examine reproductive function. Results Histopathological observations showed that both endometrial thickness and the number of glands decreased in the chemical-injured and heat-injured groups compared with the control group 7 and 14 d after treatment (P<0. 05), whereas the endometrial fibrosis area ratio was increased (P<0. 05). There were no significant differences between the mechanical damage and control groups. At 14 d after modeling, the pregnancy rate and number of uterine embryo implantations decreased in the chemical compared with the control group (P<0. 05), and the number of rabbit embryo implantations was significantly reduced (P<0. 001) in the heat- injured compared with control group. After 28 d of modeling, the chemical, infection and mechanical groups had basically recovered, but not the heat damage group. Conclusions In general, the histopathological changes observed in rabbit uteri after chemical injury modeling were more similar to the characteristics of moderate and severe IUA in humans, indicating this approach may be used as an effective model to investigate moderate and severe human IUA, and provide a basis for future in-depth study of the pathogenesis and treatment of IUA.

Abstract: Objective To study the biological characteristic values of human angiotensin-converting enzyme II (hACE2)-KI/ NIFDC mice. Methods Ten hACE2-KI/ NIFDC female mice at 4 and 8 weeks of age were tested for 11 organ coefficients, 22 physiological values and 13 biochemical values. Statistical method were used for data analysis. Results The 4-week-old mice had higher values than the 8-week-old mice in five organ coefficients ( spleen, lung, thymus, left kidney, right kidney, P<0. 01) and three physiological values (mean corpuscular volume, mean corpuscular hemoglobin, eosinophil granulocyte, P<0. 05). However, in the 4-week-old mice, the other five values were below those of the 8-week-old mice ( percentage of monocyte, alkaline phosphatase, P<0. 05; heart organ coefficients, aspartate transaminase, phosphorus, P<0. 01). Conclusions These findings may provide support for the application of hACE2-KI/ NIFDC mice .

Abstract: Objective To observe the expression levels of Farnesoid X receptor (FXR) in non-small cell lung cancer (NSCLC) tissue and cell lines, and to explore whether the role of FXR in the proliferation and invasion of NSCLC cells is related to the regulation of miR-21. Methods From 2010 to 2012, 80 cases of NSCLC tissue and matched normal tissue were collected. The expression of FXR in the NSCLC tissue and cell lines was analyzed using immunohistochemistry or Western blot. The effects of GW3965 (1 ~ 5 μmol / L) on cell viability and invasion and miR-21 levels were evaluated using CCK-8, transwell and qRT-PCR. MiR-21 was overexpressed or knocked down using transfection technology, and the effects on cell viability and invasion were evaluated. Moreover, the relationship between FXR and miR-21 was analyzed by double luciferase assay. Results Compared with the adjacent non-tumor normal tissue, the expression of FXR was significantly lower in NSCLC tissue (P<0. 001). Kaplan-Meier analysis revealed that low FXR expression predicted a worse prognosis for NSCLC patients (χ²= 4. 496, P= 0. 033). There were significant differences in clinical stage, tumor size, T stage and lymph node metastasis (N stage) between high- and low-FXR tumors ( P< 0. 05). GW3965 promoted FXR protein expression in A549 cells in a dose-dependent manner, inhibited cell proliferation and invasion and decreased miR- 21 expression. Overexpression of miR-21 significantly reversed the inhibitory effects of FXR on the growth and invasion of NSCLC cells (P< 0. 05), and miR-21 silencing significantly enhanced the inhibitory effects of FXR on the growth and invasion of NSCLC cells (P<0. 05). The double luciferase assay confirmed that FXR inhibited the proliferation and invasion of NSCLC cells by targeting miR-21. Spearman’ s test revealed a strong negative correlation between FXR and miR-21 expression in NSCLC specimens (P< 0. 001). Furthermore, Kaplan-Meier analysis revealed that a coexistence pattern of “FXR low” and “miR-21 high” predicted the worst prognosis in NSCLC patients (χ²= 8. 201, P= 0. 004). Conclusions FXR inhibits the growth of NSCLC cells by downregulating miR-21, suggesting that regulation of FXR/ miR-21 may be a potential strategy for the treatment of NSCLC.

Abstract: Objective To study the effects and mechanisms of Exendin-4 on streptozotocin-induced diabetic nephropathy in rats. Methods 60 SD healthy rats without specific pathogen level were selected. 45 rats were established to establish diabetic nephropathy model. They were randomly divided into model group, benazepril group, Exendin-4 low, medium and high dose group, and the blank group was healthy rats with 9 rats in each group. After successful modeling, benazepril group was given 10 mg / kg benazepril tablets by gavage, exendin-4 low, medium and high dose groups were injected subcutaneously with Exendin-4 4, 20 and 100 g / kg respectively, and the normal group and model group were given purified water by gavage. One week after administration, total cholesterol (TG), triglycerides (TC), high-density lipoprotein cholesterol (HDL), low-density lipoprotein cholesterol (LDL), amylase (AMY) and blood glucose (BG) were measured, hemoglobin ( HbAlc), fasting serum insulin ( FINS), body mass, renal index ( RI ), glomerular area, glomerular diameter and the expression of advanced glycation end products (AGEs), PDK1, p-Akt and p-mTOR proteins in renal tissue. Results The expression of TG, TC, LDL, BG, HbAlc, fins, ages, body mass, RI, glomerular area, glomerular diameter, PDK1, p-Akt, p-mTOR protein expression in the middle and high dose groups were lower than that in the low dose group (P<0. 05). The expressions of TG, TC, HDL, LDL, BG, HbAlc, FINS, AGEs, RI, glomerular area, glomerular diameter, PDK1, p-Akt and p-mTOR protein in Exendin-4 high dose group were lower than those in Exendin-4 medium dose group, and HDL was higher than that in Exendin-4 medium dose group. There were significant differences in TC, HDL, LDL, AMY, HDL, body mass, RI and glomerular area ( P< 0.05) Conclusions Exendin-4 can reduce inflammation levels in streptozotocin-induced diabetic nephropathy rats and improve their BG, lipid levels, and pancreas function. The mechanisms underlying these effects may be related to inhibition of the PDK1 / Akt / mTOR pathway. These findings provide a new direction for the clinical treatment of diabetic nephropathy and other inflammatory diseases.

Abstract: Objective To explore the long-term effects of uncoupling protein 1 (Ucp1) knockout on isoproterenol- induced myocardial remodeling in rats. Methods To establish the myocardial remodeling models, saline or isoproterenol was administered intraperitoneally at 30 mg / kg once per day for 3 consecutive days in wild-type (WT) and Ucp1 knockout (Ucp1^{- / -}) rats at 2 months of age. The phenotype analysis of the WT and Ucp1^{- / -}rats was analyzed using M-mode echocardiography, the serum markers of myocardial injury lactate dehydrogenase (LDH) and creatine kinase isoenzyme MB (CK-MB), and histopathological examination at 1 month after the saline or isoproterenol administration. Results In the saline group, the echocardiographic parameters, serum marker of myocardial injury CK-MB, and fibrosis were comparable between the WT and Ucp1^{- / -}rats. In the model group, Ucp1^{- / -}rats had thin-walled ventricles and decreased cardiac function, and had higher serum LDH and CK-MB levels and more serious myocardial disarray and fibrosis in the heart tissue compared with WT rats. Conclusions Ucp1 knockout aggravates isoproterenol-induced myocardial remodeling in rats. Ucp1 may be an important modifier gene of myocardial remodeling.

Abstract: Objective To investigate the effects of microRNA (miR)-137, regulated by long non-coding RNA (LncRNA) growth arrest-specific 5 ( GAS5 ), on amyloid-β ( Aβ)-induced hippocampal neuronal damage in rats. Methods Rats were divided into the control group, Aβ group, Aβ+Si-NC group, Aβ+si-GAS5 group, Aβ+si-GAS5+ anti-miR-NC group and Aβ+si-GAS5+anti-miR-137 group (n= 10 rats per group). The Y maze was used to detect learning and memory in rats. Real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) was used to detect the levels of GAS5 and miR-137 in the hippocampal CA1 area. The protein levels of B-lymphoma-2 gene (Bcl2), Bcl2-related X protein (BAX), caspase-3 were detected by Western blot and neuronal morphology was observed using Nissl staining. In addition, terminal deoxynucleotidyl transferase dUTP nick end labeling (Tunel) staining was used to observe neuronal apoptosis and the relationship between miR-137 and GAS5 was identified using double luciferase. Results Compared with the control group, the levels of GAS5 mRNA, BAX, caspase-3 protein, neuronal mortality, and neuronal apoptosis rate were higher in the Aβ and Aβ+Si-NC groups (P<0. 05), whereas the proportion of correct response times, the proportion of learning times, the levels of miR-137 and Bcl2 protein were lower (P<0. 05). Compared with the Aβ and Aβ+Si-NC groups, the levels of GAS5 mRNA, BAX, caspase-3 protein, neuronal mortality and neuronal apoptosis rate were lower in the Aβ+si-GAS5 and Aβ+si-GAS5+anti miR-NC groups (P<0. 05), whereas the proportion of correct response times, the proportion of learning times, the levels of miR-137 and Bcl2 protein were higher (P<0. 05). In addition, compared with the Aβ+si-GAS5 and Aβ+si-GAS5+anti-miR-NC groups, the levels of GAS5 mRNA, BAX, caspase-3 protein, neuronal mortality, and neuronal apoptosis rate were higher in the Aβ + si-GAS5 + anti-miR-137 group ( P< 0. 05), whereas the proportion of correct response times, the proportion of learning times, the levels of miR-137 and Bcl2 protein were lower (P<0. 05). It was also confirmed that there was a target site between miR-137 and GAS5. Conclusions Interference of GAS5 can upregulate miR-137, thus protecting against Aβ-induced hippocampal neuronal damage and alleviating the process of neuronal apoptosis. In contrast, further downregulation of miR-137 can reverse this process.

Abstract: Objective To investigate the changes in renal functions and the renin-angiotensin system (RAS) at different stages in endotoxemia rats, to provide data to support its pathological mechanism. Methods Ninety rats were randomly divided into nine groups: the control group (n= 10) and the model group at eight timepoints (2, 4, 8, 12, 24, 48, 72 and 168 h, n= 10). Rats in the model group were prepared for the sepsis model by a tail vein injection of lipopolysaccharides, while those in the control group were injected with an equal volume of pyrogen-free water. Samples were collected from the animals in the control group at 8 h after injection and from animals in the model group at each timepoint. The circulating endotoxin, nitric oxide (NO), endothelin ( ET), nitric oxide synthase (NOS), angiotensin converting enzyme (ACE), angiotensin (Ang), renin-angiotensin (PRA), serum creatinine and blood urea nitrogen levels were detected by enzyme-linked immunosorbent assay, the protein expression of AngII type 1 receptor (AT1R) and Ang II type 2 receptor ( AT2R) in the kidney was measured using Western blot assay. Results After lipopolysaccharide injection, the circulating endotoxin, endothelin, NO, NOS, ACE, AngⅡ, serum creatinine, blood urea nitrogen, AT1R and AT2R levels were all increased in the model group. The kidney coefficient, serum endotoxin, serum creatinine, blood urea nitrogen and kidney AT1R and AT2R were higher in the model group than in the control group at 2 ~ 168 h after injection. In addition, the serum levels of endothelin, NOS and AngⅡ were higher in the model group than in the control group at 2~ 48 h after injection and the NO level in the model group was higher than that in the control group at 4 ~ 48 h after injection. Moreover, the ACE level in the model group was higher than that in the control group at 2 ~ 24 h after injection. All differences were significant ( P< 0. 05). Conclusions RAS may be involved in kidney injury caused by endotoxemia.

Abstract: Objective Herba Patriniae is a root grass of perennial herbaceous plants in the family Caprifoliaceae, with various pharmacological activities. This study was conducted to investigate the protective effect of Herba Patriniae on paracetamol-induced liver injury in rats. Methods Rats were intravenously injected with acetaminophen and then randomly divided into the control, model, low-dose Herba Patriniae, medium-dose Herba Patriniae, high-dose Herba Patriniae and bifendatatum groups. The low-, medium- and high-dose Herba Patriniae groups were gavaged with 4, 8 and 16 g / kg Herba Patriniae once per day, respectively. The bifendatatum group received 0. 3 g / kg bifendatatum, and the control and model groups were given the same amount of normal saline for 30 consecutive days. The liver index was calculated. Hematoxylin and eosin staining was used to detect the degree of pathological injury to the liver tissue. Serum ALT, AST and ALP levels were detected using kits. Tumor necrosis factor-α ( TNF-α), interleukin-6 ( IL-6) and interleukin-1β ( IL-1β) were determined via enzyme-linked immunosorbent assay. Superoxide dismutase (SOD), MDA and GSH-Px contents in the liver homogenate were detected using kits. Results Compared with those of the control group, the liver index of the model group was significantly increased, the AST, ALT, ALP, TNF-α, IL-6 and IL-1β levels were significantly increased, the SOD and GSH-Px contents were significantly reduced, and the MDA content was significantly increased (all P<0. 05). Compared with those of the model group, the liver indexes of the Herba Patriniae and bifendatatum groups were significantly reduced, the degree of pathological damage was significantly attenuated, the AST, ALT, ALP, TNF-α, IL-6 and IL-1β levels were significantly decreased, the SOD and GSH-Px contents were significantly increased, and the MDA content was significantly decreased (all P<0. 05). Conclusions Herba Patriniae protected rats from paracetamol-induced liver injury by alleviating pathological injury to the liver tissue, inhibiting the release of inflammatory factors and relieving oxidative stress.

Abstract: Objective To study the intervention effect of vascular endothelial growth factor (VEGF) inhibitors on diabetic retinopathy model rats and the hemodynamics of central retinal artery (CRA). Methods Of 40 specific-pathogen- free healthy male rats, 10 were randomly selected as the control group. The remaining 30 rats were successfully modeled and randomly divided into the model group ( normal saline), sterile citric acid group ( sterile citric acid) and VEGF inhibitor group ( bevacizumab) ( n= 10). The minimum diastolic velocity ( EDV) of the CRA was measured. Results Compared with the model group, the levels of EDV (31. 53±8. 42 / 47. 46± 10. 29 / 90. 26± 16. 55 mm/ s, F/ P= 14. 956 / 0. 001), peak systolic velocity ( PSV; 58. 92± 11. 37 / 84. 16± 14. 52 / 155. 57± 18. 67 mm/ s, F/ P= 21. 116 / 0. 001) and adiponectin were increased in the aseptic citric acid and VEGF inhibitor groups. In addition, the resistivity index (RI; 1. 12±0. 32 / 0. 85±0. 20 / 0. 68±0. 18, F/ P= 7. 046 / 0. 001), pulsatility index (PI; 1. 35±0. 35 / 0. 91±0. 22 / 0. 69±0. 12, F/ P= 8. 861 / 0. 001 ), hemorheological indicators, fasting blood glucose, VEGF, intercellular adhesion molecule 1 (ICAM-1), interleukin-1 (IL-1) and expression levels of Ras, Raf and extracellular signal-regulated kinase (ERK) were significantly decreased (P<0. 05). Compared with the aseptic citric acid group, the levels of EDV (47. 46±10. 29 / 90. 26± 16. 55 mm/ s, T/ P= 6. 945 / 0. 001), PSV (84. 16±14. 52 / 155. 57±18. 67 mm/ s, T/ P= 9. 548 / 0. 001) and adiponectin were increased in the VEGF inhibitor group. Furthermore, the RI ( 0. 85± 0. 20 / 0. 68± 0. 10, T/ P= 2. 404 / 0. 027), PI (0. 91±0. 22 / 0. 69±0. 12, T/ P= 2. 776 / 0. 012), hemorheological indicators, fasting blood glucose, VEGF, ICAM-1, IL- 1 and expression of Ras, Raf-1, and ERK were significantly decreased (P<0. 05). Conclusions VEGF inhibitors can improve the CRA hemodynamics and hemorheology indices of diabetic retinopathy model rats, reduce the levels of VEGF, ICAM-1 and IL-1β and inhibit angiogenesis.

Abstract: Objective To optimize the experimental conditions for the determination of porcine endogenous retrovirus (PERV) copy number in the porcine genome using a droplet digital polymerase chain reaction ( ddPCR) technique. Methods TaqMan probe duplex-ddPCR was used to determine the copy number of PERV, glyceraldehyde 3-phosphate dehydrogenase ( GAPDH) and transferrin receptor protein 1 ( TFRC) were adopted as the internal reference genes. To determine the optimal annealing temperature and PCR cycle number, we compared multiple conditions. In addition, the suitable range of template amount was estimated by examining the serially diluted plasmid and the pig cell genome. Genomic DNA isolated from pig cell lines or Bama minipig tissue was used to determine the copy number of PERV using ddPCR-and quantitative PCR-based method, which demonstrated the repeatability of different method. Results The optimal annealing temperature for the ddPCR-based method to measure PERV copy number was 59. 9℃ , and the optimal number of PCR cycles was 50. The suitable template genome concentration ranged from 1. 25 to 7. 5 ng. The variation of coefficients for PERV copy number in the porcine genome ranged from 0. 44% to 8. 29%. Conclusions A method for analyzing the PERV copy number based on the ddPCR technique was established, which improved the accuracy and repeatability compared with a traditional quantitative PCR-based detection approach. This method provides a sensitive approach and is a reliable basis for the determination of PERV copy number in donor animal genomes for xenotransplantation studies.

Abstract: Objective To explore the effects of Oridonin on the proliferation and apoptosis of hepatic stellate cells and their molecular mechanism. Methods Different concentrations of Oridonin were used to intervene in hepatic stellate cells ( HSC-T6 ) for 48 h. The methyl thiazolyl tetrazolium (MTT) assay and flow cytometry were used to detect proliferation and apoptosis in HSC-T6 cells. Real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot were used to detect the expression changes of ubiquitin-conjugating enzyme 9 (UBC9). The transfection of UBC9 small interfering RNA (si-UBC9) downregulated the expression level of UBC9 in HSC-T6 cells. Changes in cell proliferation and apoptosis were also evaluated using the aforementioned method. Results After Oridonin intervention, the proliferation inhibition rate and apoptosis rate of HSC-T6 cells were significantly increased, the expression of UBC9 was significantly decreased and there was a significant dose-dependent relationship (P<0. 05). After downregulating UBC9 expression, the proliferation inhibition rate and apoptosis rate of HSC-T6 cells were significantly increased and UBC9 expression was significantly decreased ( P< 0. 05). The overexpression of UBC9 was able to reverse the effects of Oridonin on the proliferation and apoptosis of HSC-T6 cells ( P< 0. 05). Conclusions Oridonin can inhibit proliferation and induce apoptosis in hepatic stellate cells. The underlying mechanism is related to the downregulation of UBC9 expression.

Abstract: Objective To investigate the role and mechanisms of platelet microparticles ( PMP) sin vascular endothelial injury of the diabetic aorta. Methods SD rats were randomly divided into the normal (NC) and diabetic (DM) groups. PMPs were isolated and purified. Next, primary cultured endothelial cells were transfected with microRNA (miR)-4306 to detect PMPs in vascular endothelial cells. The effects of activity and the effects on the miR-4306 / vascular endothelial growth factor A ( VEGFA) / extracellular signal-regulated kinase ( ERK) 1 / 2 / nuclear factor κB ( NF-κB) signaling pathway were evaluated. Results The positive expression and apoptosis rates of endothelial nitric oxide synthase (eNOS) and caspase-3 in the DM group were significantly different from those in the NC group (P<0. 05 or P<0. 01). The relative expression of miR-4306 in the mimic-M group and inhibitor-M group was significantly different from that in the miR-M group (t= 3. 821, 4. 597, P<0. 05 or P<0. 01). Moreover, the relative expression of miR-4306 in the mimic-M group was significantly different from that in the inhibitor-M group (P<0. 01). The relative expression of miR-4306 in M group was significantly different from that in Ctrl group (P<0. 01). In addition, the relative expression of VEGFA, NF- κBp65, p-IκBα and p-ERK in the mimic-M group and inhibitor-M group was significantly different from that in M group(P<0. 05 or P<0. 01). The relative expression of VEGFA, NF-κBp65, p-IκBα and p-ERK in the M group was significantly different from that of Crtl group (P<0. 05 or P<0. 01). Conclusions PMPs may cause endothelial damage in the diabetic aorta, the underlying mechanism may be related to the activation of miR-4306 / VEGFA/ ERK1 / 2 / NF-κB signaling.

Abstract: Objective To explore the optimal method of producing a streptozotocin-induced diabetic nephropathy C57BL/ 6 mice model by investigating a molded rate of single high-dose and multiple low-dose streptozotocin injections. Methods Male C57BL/ 6J mice ( n= 48) were randomly divided into three groups: the normal group and two diabetic nephropathy groups. Mice in one model group (the single high-dose group) were intraperitoneally administered 150 mg / kg streptozotocin after having a high fat / high sucrose diet. The other model group ( the multiple low-dose group ) was intraperitoneally administered 50 mg / kg streptozotocin for 4 consecutive days after having a high fat / high sucrose diet. The fasting blood glucose levels, body weight, water intake, 24 h urine protein, renal function and renal pathological changes of the three groups were measured. Results The body weight of the single high-dose group increased stably after streptozotocin injection, while the body weight of the multiple low-dose group decreased by inches after streptozotocin injection. Moreover, the fasting blood glucose levels, water intake and 24 h urine protein increased stably in the multiple low-dose group from week 2 to week 10. The levels of fasting blood glucose, 24 h urine protein, total cholesterol, triglycerides, low density lipoprotein, serum creatinine, blood urea nitrogen and kidney / body weight ratio were significantly higher in the multiple low-dose group than in the single high-dose group ( P< 0. 05) and the change in pathology was apparent. Conclusions The multiple low-dose streptozotocin injection protocol is a reasonable method for inducing mouse diabetic nephropathy.

Abstract:The outbreak of Corona Virus Disease 2019 brought great challenges to the allocation of health human resources in medical and health institutions. Not only should the hospitals undertake the medical treatment and on-site rescue of patients caused by emergencies, but also compact defenses to minimize the spread of virus in medical institutions, avoid cross infection, and take daily medical services into account. This study aims to summarize the hospital’ s human resources emergency management strategies in a comprehensive public hospital during Corona Virus Disease 2019, in order to provide reference for other hospitals to deal with public health emergencies.

Abstract:Alcohol can induce brain dysfunction, and gut microbiota can mediate the effects of alcohol on brain injury through the gut-brain axis. At present, the mechanisms of alcohol-mediated brain injury through the microbiota-gut- brain axis mainly include inflammation, endotoxins and tryptophan metabolism. Exploring the interactions between alcohol and the microbiota-gut-brain axis will be helpful for understanding the effects of alcohol on the brain, and will provide a new way to explore the pathogenesis of alcohol-induced brain injury.

Abstract:Bone marrow mesenchymal stem cells (BMSCs) are non-hematopoietic stem cells in the bone marrow that have the advantages of multidirectional differentiation and low immunogenicity. Although liver transplantation is an effective treatment for end-stage liver disease, its clinical application is severely limited by the shortage of donor liver sources and rate of immune rejection. The differentiation of BMSCs into hepatocytes in vitro using different induction method is applied in the treatment of advanced liver disease, and represents a new treatment method for liver injury. This article summarizes the isolation culture, and the common method and mechanisms for inducing BMSC differentiation into hepatocytes that have been described in the literature in recent years. A clear understanding of these processes will provide a strong theoretical basis for future clinical applications of BMSCs in the treatment of end-stage liver disease.

Abstract:Polydatin is a monocrystalline compound isolated from Polygonum cuspidatum Sieb. et Zucc. (Polygonaceae). Modern pharmacological studies have indicated that polydatin has a variety of pharmacological activities, such as anti-thrombosis, anti-arteriosclerosis, improvement of myocardial damage, anti-shock and regulation of glucose and lipid metabolism. In recent years, studies have reported that polydatin also has anti-liver cancer and anti-liver fibrosis effects and protects hepatocytes and improves hepatocyte steatosis. This article reviews the pharmacological effects and mechanisms of polydatin in liver disease to provide a reference for its follow-up research, development and use.

Abstract:As one of the most common malignant tumors, lung cancer is the main cause of cancer death. Early diagnosis is the key to improving the quality of life and prognosis of patients with lung cancer and the detection of tumor markers is important for the early diagnosis of lung cancer. The early appearance and high concentrations of tumor markers in bronchoalveolar lavage fluid are helpful for the early diagnosis of lung cancer. This article reviews the research status of tumor markers in bronchoalveolar lavage fluid in recent years and provides a reference for the early diagnosis of lung cancer.

Abstract:Leukemia is a kind of hematological malignance characterized by the clonal expansion of leukemic cells and has a high lethality. Paclitaxel is a natural anticancer drug that has been widely used in chemotherapy against multiple malignancies because of its unique anticancer effects. However, there are relatively few studies of paclitaxel use in the treatment of leukemia. This paper reviewed the researchs of paclitaxel in the treatment of leukemia.