Hyaluronic Acid Levels Research Articles

Helix-Loop-Helix Factor Id3 (Inhibitor of Differentiation 3): A Novel Regulator of Hyaluronan-Mediated Adipose Tissue Inflammation.

The aim of this study was to unravel mechanisms whereby deficiency of the transcription factor Id3 (inhibitor of differentiation 3) leads to metabolic dysfunction in visceral obesity. We investigated the impact of loss of Id3 on hyaluronic acid (HA) production by the 3 HAS isoenzymes (HA synthases; -1, -2, and -3) and on obesity-induced adipose tissue (AT) accumulation of proinflammatory B cells. Approach and Results: Male Id3-/- mice and respective wild-type littermate controls were fed a 60% high-fat diet for 4 weeks. An increase in inflammatory B2 cells was detected in Id3-/- epididymal AT. HA accumulated in epididymal AT of high-fat diet-fed Id3-/- mice and circulating levels of HA were elevated. Has2 mRNA expression was increased in epididymal AT of Id3-/- mice. Luciferase promoter assays showed that Id3 suppressed Has2 promoter activity, while loss of Id3 stimulated Has2 promoter activity. Functionally, HA strongly promoted B2 cell adhesion in the AT and on cultured vascular smooth muscle cells of Id3-/- mice, an effect sensitive to hyaluronidase. Our data demonstrate that loss of Id3 increases Has2 expression in the epididymal AT, thereby promoting HA accumulation. In turn, elevated HA content promotes HA-dependent binding of B2 cells and an increase in the B2 cells in the AT, which contributes to AT inflammation.

Ozone exposure promotes pyroptosis in rat lungs via the TLR2/4-NF-κB-NLRP3 signaling pathway

ObjectiveOzone has become a major air pollutant in recent years, which leading to a variety of lung diseases. This study aimed to explore the mechanisms of pyroptosis and related signaling pathways in ozone-induced lung injury. MethodsWe exposed 120 Wistar rats to ozone, 20 in each group (half male and half female). Ozone exposure concentrations were 0, 0.12, 0.5, 1.0, 2.0 and 4.0 ppm for 6 h. At the same time, we isolated and cultured type I alveolar epithelial cells, then intervened with high mobility group box 1 protein (HMGB1), hyaluronic acid (HA) and Toll-like receptors 2/4 (TLR2/4) inhibitor. In animal experiments, histopathological experiments, TUNEL, ELISA and biochemical indicators were performed. RT-qPCR and western blot experiments assay were used to detect the expression changes of key factors in relevant signal pathways in vivo and in vitro. ResultsAfter acute ozone exposure, the levels of lung cell injury indicators in bronchoalveolar lavage fluid (BALF), as well as the levels of inflammatory factors in BALF, blood, and lung tissue were significantly increased. Male rats were more sensitive to ozone exposure. Low-concentration ozone exposure caused mild interstitial inflammation in rat lung tissue. Severe inflammation and pulmonary edema appeared with increases in concentration. ELISA results in BALF showed that HMGB1 and HA expressions increased gradually with the increase of ozone exposure concentration. RT-qPCR and Western blot showed that when ozone concentrations increased above 0.5 ppm, the expression of nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing 3 (NLRP3), cleaved caspase-1, and N-gasdermin D (N-GSDMD) in the lung tissue increased significantly, suggesting that ozone exposure induces pyroptosis. At the same time, it was found that ozone exposure activated the nuclear factor kappa B (NF-κB) signal pathway, and increased the mRNA expressions of Toll-like receptors TLR2/4. The results of cell experiments showed that after the addition of HMGB1 and HA, the expression of NF-κB and pyroptosis related indexes increased in type I alveolar epithelial cells, while the corresponding expression decreased after the addition of TLR2/4 inhibitors. ConclusionOzone exposure causes lung injury in a dose- and gender-dependent manner, and is more severe in males. When injured, the levels of HMGB1 and HA in BALF increased, which interact with TLR 2/4 to activate the downstream NF-κB signaling pathway. Further activating the NLRP3 inflammasome complex and regulating the ozone-induced pyroptosis.

Study of serum traditional and nontraditional biomarkers in Rheumatoid arthritis patients

The study of biomarkers in Rheumatoid arthritis (RA) is highly indispensable to understand mechanisms of pathogenesis, diagnosis, prognosis, and treatment of disease. The role of traditional biomarkers like Anti-CCP, RF, and inflammatory markers like ESR and CRP is well established. In this study, we aimed to measure nontraditional biomarkers like Hyaluronic acid (HA), Cartilage oligomeric matrix protein (COMP), and Osteocalcin in the serum of RA patients and also to establish an association with traditional markers. It was a cross-sectional study involving 152 RA patients based on the 1987 ACR criteria for the diagnosis of RA and 68 age‑ and sex-matched healthy controls. After the clinical examination, traditional markers were assessed to measure the disease activity along with non traditional markers in RA patients. All the values were expressed as median (25th–75th percentile). In our study, there was a significant increase in serum HA levels in RA patients compared to healthy controls (p < 0.03), whereas no significant difference in serum COMP and osteocalcin levels. The traditional inflammatory markers were significantly increased in RA patients than controls with (p < 0.001). The serum HA levels were significantly correlated with traditional markers in RA patients. Conclusion: Significant increase in serum HA level in RA patients indicating synovial inflammation, but there were no notable changes in COMP and osteocalcin level in serum presuming the combination of these markers may be useful along with traditional markers in the different stages of RA.

Analysis of novel serum markers of fibrosis and angiogenesis in patients with alcoholic liver cirrhosis.

Alcohol consumption causes acute and chronic liver injury. The clinical forms of alcohol liver disease (ALD) include steatosis, hepatitis, cirrhosis, and hepatocellular carcinoma (HCC) associated with liver cirrhosis. The aim of the study was to determine the levels of novel markers of fibrogenesis and angiogenesis in patients with alcoholic liver cirrhosis. Serum levels of angiopoietin-like peptide 4 (ANGPTL-4), asialoglycoprotein receptor 1 (ASGP-R1), and S100 calcium-binding protein A8 (S100A8) were assessed. Levels of hyaluronic acid (Hyal) and collagen IV (Coll IV) werealso determined at various stages of alcoholic liver cirrhosis. The study group consisted of 72 patients with alcoholic liver cirrhosis, while the control group included 22 healthy subjects without a history of alcohol abuse. The degree of liver cirrhosis was evaluated according to the Pugh-Child criteria (Pugh-Child score). Based on thse scores, patients were assigned to one of three groups: Pugh-Child (P-Ch) A - 21 with stage A, P-Ch B - 23 with stage B and P-Ch C - 28 with stage C liver cirrhosis. Serum levels of markers were determined using ELISA. The study findings demonstrated higher levels of ANGPTL-4, ASGP-R1, S100A, hyaluronic acid and serum collagen IV in the group of patients with alcoholic liver cirrhosis, compared to the control group. Furthermore, their levels increased with the progression of alcoholic liver cirrhosis. The biomarkers analysed in the study may be useful for diagnosis and prognosis in patients with alcoholic liver cirrhosis.

Non-alcoholic fatty liver disease: non-invasive markers and their importance in diagnosis

Objective — to evaluate the validity of serological markers in the diagnosis of non‑alcoholic fatty liver disease (NAFLD).Materials and methods. The investigation involved 122 patients with NAFLD aged 19 to 74, including 57 women (46.7 %) and 65 men (53.3 %). The following investigative methods have been used: anthropometric measurements (height, weight, waist circumference, BMI); ultrasound examination of the liver; laboratory (determination of lipid spectrum, insulin resistance, ALT, AST, fibrinogen, free oxyproline, oxyproline protein‑bound, hyaluronic acid, haptoglobin, apolipoprotein A1, a2‑macroglobulin, fibronectin); morphological (puncture biopsy of the liver with histological examination of biopsies); statistical (descriptive statistics, correlation, linear regression and ROC‑analysis).Results. The direct correlation between the degree of steatosis and level of protein‑bound oxyproline (r = 0.22; p < 0.05), the ratio of oxyproline protein‑bound/oxyproline free (r = 0.27; p < 0.05), the level of triglycerides (r = 0.33; p < 0.05), the level of VLDL (r = 0.29; p < 0.05). The direct correlation of moderate strength has been established between the degree of the activity of inflammatory process and levels of hyaluronic acid, a2‑macroglobulin and fibronectin (r = 0.54, 0.67 and 0.55, respectively; p < 0.05 for all) and a moderate inverse relationship for apo‑lipoprotein A1 (r = – 0.56; p < 0.05), haptoglobin (r = – 0.33; p < 0.05) and albumin (r = – 0.45; p < 0.05). Evaluation of correlations showed a direct relationship between the stage of fibrosis and the level of hyaluronic acid, a2‑macro­globulin and fibronectin (r = 0.72, 0.93 and 0.71 at p < 0.05) and a weak relationship with total bilirubin (r = 0.26; p < 0.05). Moderate feedback (r = – 0.61 and ‑0.35 — 0.32; p < 0.05, respectively) was found between apolipoprotein A1, haptoglobin, albumin and fibrosis stage.Conclusions. In patients with steatohepatitis, high diagnostic value was determined for non‑invasive markers of fibrosis: a2‑macroglobulin, apolipoprotein A1, hyaluronic acid and fibronectin for which the area under the ROC curve for minimal fibrosis was not less than 0.75, but more than moderate 0.9. Patients with liver cirrhosis showed a high level of sensitivity (96.4 — 100.0 %) and specificity (75.0 — 100.0 %) of these markers.

Association of CX3CL1 and CX3CR1 Expression with Liver Fibrosis in a Mouse Model of Schistosomiasis.

Immunopathological mechanisms of schistosomiasis, a debilitating parasitic disease, are still unclear. In this study, we investigated the involvement of CX3C chemokine ligand 1 (CX3CL1) and its sole receptor CX3CR1 in the development of liver fibrosis in schistosomiasis. The animal model of schistosomiasis was established by infection of C57BL/6 mice with Schistosoma japonicum cercariae; mice injected with carbon tetrachloride (CCl4) were used as positive control of liver injury. After 4 and 8 weeks, the degree of liver lesions was assessed by hematoxylin and eosin staining, serum levels of hyaluronic acid (HA) were analyzed by a chemiluminescence immunoassay, liver fibrosis was evaluated by immunohistochemistry analysis of α-smooth muscle actin (α-SMA) expression, and CX3CL1 and CX3CR1 expression in the liver was measured by immunohistochemistry and real-time PCR. The results showed that at 8 weeks after Schistosoma infection, serum HA levels were increased and α-SMA-expressing cells appeared in the liver, indicating fibrogenesis. CX3CL1- and CX3CR1-positive cells were observed in the outer layer of granulomas formed around Schistosoma eggs in liver tissues, which was consistent with the significant upregulation of hepatic CX3CL1 and CX3CR1 mRNA expression at 4 and 8 weeks post-infection. Furthermore, correlation analysis revealed positive association between CX3CL1 and CX3CR1 expression and serum HA levels at 8 weeks post-infection, indicating a link between fibrogenesis and the CX3CL1/CX3CR1 axis in schistosomiasis. In conclusion, our data suggest the involvement of CX3CL1 and CX3CR1 in the progression of liver fibrosis caused by Schistosoma infection.

18294 Topical treatment with kiwi-derived yeast extract increases hyaluronic acid resulting in improved skin hydration and clinically perceived facial radiance

Background: Hyaluronic acid (HA) is a key molecule involved in maintaining skin structure and moisture, synthesized by hyaluronic acid synthase 2 enzyme (HAS2). However, HAS2 gene expression and activity decrease with age, resulting in declining HA levels, loss of skin radiance and decreased skin firmness. We investigated the effect of a topical Kiwi-derived Yeast Extract (KDYE) on HA and HAS.

Modeling the effects of hyaluronic acid degradation on the regulation of human astrocyte phenotype using multicomponent interpenetrating polymer networks (mIPNs)

Hyaluronic acid (HA) is a highly abundant component in the extracellular matrix (ECM) and a fundamental element to the architecture and the physiology of the central nervous system (CNS). Often, HA degradation occurs when an overreactive inflammatory response, derived from tissue trauma or neurodegenerative diseases such as Alzheimer’s, causes the ECM in the CNS to be remodeled. Herein, we studied the effects of HA content as a key regulator of human astrocyte (HAf) reactivity using multicomponent interpenetrating polymer networks (mIPNs) comprised of Collagen I, HA and poly(ethylene glycol) diacrylate. The selected platform facilities the modulation of HA levels independently of matrix rigidity. Total astrocytic processes length, number of endpoints, the expression of the quiescent markers: Aldehyde Dehydrogenase 1 Family Member L1 (ALDH1L1) and Glutamate Aspartate Transporter (GLAST); the reactive markers: Glial Fibrillary Acidic Protein (GFAP) and S100 Calcium-Binding Protein β (S100β); and the inflammatory markers: Inducible Nitric Oxide Synthase (iNOS), Interleukin 1β (IL-1β) and Tumor Necrosis Factor Alpha (TNFα), were assessed. Cumulatively, our results demonstrated that the decrease in HA concentration elicited a reduction in the total length of astrocytic processes and an increase in the expression of HAf reactive and inflammatory markers.

In Vitro Anti-Inflammatory and Skin Moisturizing Properties of Pilea martini (Levl.) Hand.-Mazz. Extracts

The in vitro anti-inflammatory and skin moisturizing activities of Pilea martini (Levl.) Hand.-Mazz. were investigated on lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages and human immortalized keratinocytes. Chromatographic analysis was performed to identify the chemical composition of the extracts. Pilea martini extracts significantly suppressed LPS-induced nitric oxide, prostaglandin E2, interleukin 6, and tumor necrosis factor α production in dose-dependent manners. In addition, the extracts inhibited LPS-induced inducible nitric oxide synthase and cyclooxygenase-2 proteins and their mRNA expression through causing a downregulation of nuclear factor-κB, activator protein 1, and mitogen-activated protein kinase signaling cascades. The extracts increased the production of hyaluronic acid levels and enhanced the expression levels of both filaggrin and serine palmitoyltransferase regulation. Liquid chromatography-mass spectroscopy analysis showed that the extracts contained 6 different compounds (malic acid, tryptophan, chlorogenic acid, caffeic acid, p-coumaric acid, and isoquercetin) that may contribute to their bioactivities. Taken together, Pilea martini extract showed remarkable promise as an anti-inflammatory and moisturizing agent.

Association of human platelet antigens polymorphisms with the levels of serum fibrosis marks in chronic hepatitis C patients

BackgroundHost genetic polymorphisms influence the fibrosis progression of chronic hepatitis C (CHC) patients. Previous studies have shown the association of human platelet antigens (HPAs) polymorphisms with CHC. However, little is known regarding the association of HPAs polymorphisms with the fibrosis progression of CHC. The aim of this study was to determine the association of HPA -2, -3, -5 and -15 polymorphisms with the levels of serum fibrosis marks in CHC patients. MethodsThe HPA -2, -3, -5 and -15 were genotyped by 5ˊ-nuclease assay in 211 CHC patients, while the serum concentration of hyaluronic acid (HA), collagen IV (CIV), amino-terminal pro-peptide of type III procollagen (PIIINP), and laminin (LN) from the same samples were measured by time resolved fluorescence immunoassay. ResultsThe level of serum LN was significantly lower in CHC patients with HPA-15aa genotype compared to those with HPA-15ab/bb (P = 0.032) but did not differ among HPA-2, -3 and -5 genotypes. There were no difference in HA, CIV and PIIINP levels among HPA-2, -3,-5 and -15 genotypes. ConclusionsThis study demonstrates that HPA-15 aa polymorphism is associated lower serum LN in CHC, which suggests that HPA -15 aa may be involved in the fibrosis progression of CHC.

Long-term results of additional pulmonary blood flow with bidirectional cavopulmonary shunt

ObjectiveWe evaluated additional pulmonary blood flow at the time of bidirectional cavopulmonary shunt and its effects on the Fontan procedure and long-term outcome of Fontan circulation and liver function.MethodsWe included 22 patients (16 boys, 6 girls) having undergone bidirectional cavopulmonary shunt with additional pulmonary blood flow between April 2002 and January 2016. Mean age and body weight were 20 ± 13 months and 7.5 ± 6.5 kg, respectively. We retrospectively evaluated the patients’ clinical data, including cardiac catheterization data, liver function, and liver fibrosis markers.ResultsAll patients were alive with a New York Heart Association status of I at the long-term follow-up. Changes between pre-bidirectional cavopulmonary shunt and 101 months after the Fontan procedure included the following: the cardiothoracic ratio of chest X-ray decreased from 52.2 ± 3.9% to 41.8 ± 5.9% (p < 0.001); systemic ventricle end-diastolic pressure decreased from 11.4 ± 3.2 mmHg to 6.9 ± 3.6 mmHg (p < 0.001); and the pulmonary artery index decreased from 485.1 ± 272.3 to 269.5 ± 100.5 (p = 0.02). Type IV collagen, hyaluronic acid, and procollagen levels increased over the normal range 116 months after the Fontan procedure.ConclusionsThe additional pulmonary blood flow at the time of bidirectional cavopulmonary shunt may contribute to pulmonary arterial growth at the Fontan procedure with low pulmonary arterial resistance and without ventricle volume overload. The Fontan circulation was well-maintained at the long-term follow-up, while liver fibrosis markers were above their normal values.

Correlation analysis of the prognostic value of serum hyaluronic acid for breast cancer patients

Objective: To explore the correlation between preoperative serum hyaluronic acid (HA) level and prognosis of breast cancer patients. Methods: The 98 patients with breast cancer who underwent surgical treatment in the Oncology Department of Sir Run Run Shaw Hospital, Zhejiang University School of Medicine from January 2004 to November 2014 in a historical cohort were included, aged (52.5±9.4) years.The preoperative serum HA contents of the patients were detected. According to the median of 53.7 μg/L, the patients were divided into high and low groups with 49 patients in each group.The χ2 test was used to analyze the correlation between the serum HA content and the general clinical data of the patients, and the Kaplan-Meier method, Log-rank test and multivariate Cox regression model wereusedto analyze the correlation between HA content and patients' survival. Results: The percentages of patients with high HA levels in menopause and non-menopause patientswere 55.7% and 40.5%, respectively; in progesterone receptor (PR) positive and negative patients were 54.1% and 43.2%, respectively; in estrogen receptor (ER) positive and negative patients were 45.7% and 60.7%, respectively; in Ki-67 positive and negative patients were 55.6% and 43.2%, respectively; in the tumor size stage TⅠ, TⅡ, TⅢ, and TⅣ patients were 50.0%, 41.7%, 72.7%, and 1/1, respectively; in lymph node metastasis and non-metastasispatients were 45.7% and 53.8%, respectively. There was no significant correlation between the level of HA and the menopausal status, the expressions of PR, ER and Ki-67, tumor size, and lymph node metastasis in breast cancer patients (χ²=2.128, 1.086, 1.800, 1.485, 4.273, 0.656, P>0.05). Patients with high HA levels accounted for 30.9% of patients aged 52 years or less and 74.4% of patients older than 52 years (χ²=18.274, P=0.000); 43.4% of patients with early TNM and 72.7% of patients with advanced TNM (χ²=5.861, P=0.015); 45.2% of patients without distant metastasis and 78.6% of patients with distant metastasis (χ²=5.333, P=0.023); 38.1% of Her-2 negative patients and 58.9% of Her-2 positive patients(χ²=4.167, P=0.041); and the median survival of patients with high HA levels was 70 months, which was shorter than 83 months for patients with low HA levels (χ²=6.799, P=0.007). Therefore, ahigh HA content predicts an older age, a later tumor stage, higher risk of distant metastasis, positive expression of Her-2 and shorter survival. Multivariate Cox regression model analysis suggested that high levels of serum HA may be a risk factor for patients' survival, with HR (95%CI) value of 9.98 (1.16-85.88) and P value of 0.036. Conclusion: The high level of preoperative serum HA has a certain correlation with the poor prognosis of breast cancer patients.

MicroRNA-146a downregulates the production of hyaluronic acid and collagen I in Graves' ophthalmopathy orbital fibroblasts.

The present study aimed to investigate the effect of microRNA (miR)-146a on the secretion of hyaluronic acid (HA) and collagen I in Graves' ophthalmopathy (GO) orbital fibroblasts, and identify potential novel targets for the clinical treatment of GO. Orbital fibroblasts were extracted from orbital connective tissue, and primary cells were identified via immunohistochemistry. The levels of HA and collagen I in orbital fibroblasts of non-GO controls and patients with GO were examined via reverse transcription-quantitative PCR (RT-qPCR). miR-146a was overexpressed or inhibited in primary orbital fibroblasts via lentiviral infection, and the levels of HA and collagen I following miR-146a overexpression or inhibition were detected via ELISA and RT-qPCR. The results indicated that the mRNA expression of HA and collagen I was higher in orbital fibroblasts from patients with GO compared with the non-GO cohort. Overexpression of miR-146a reduced, and inhibition of miR-146a increased the production of HA and collagen I in GO orbital fibroblasts. In conclusion, it was demonstrated that miR-146a downregulated the secretion of HA and collagen I in GO orbital fibroblasts in vitro, which may affect glycosaminoglycan aggregation and collagen deposition in GO.

The influence of different concentration of proFLEX® INTRA on clinical progression and dynamics of laboratory test values in different age patients with knee osteoarthrosis

Introduction: application of hyaluronic acid gel for intraarticular administration is actual topic of modern orthopaedic concerning to its therapeutical efficacy. Objective: to assess the influence of different concentrations of proFLEX® INTRA on the synovial fluid and biochemical blood markers in patients with the 2nd knee osteoarthritis stage in the different age groups. Materials and methods: 30 patients with Kellgren–Lawrence stage II knee osteoarthritis were examined. The first group consisted of 15 patients aged 40–50 years, in the second group the average age of patients was 50–70 years. Each group was divided into 3 subgroups (a, b, c) of 5 patients. In each group the first subgroup of patients received intra-articular 1 injection of proFLEX® INTRA 10 mg/ml 2.0 ml, І-b, ІІ-b — 12 mg/ml 2.5 ml, І-с, ІІ-с — 20 mg/ml 3.0 ml. Results: an inflammatory-dystrophic process in the knee joint of patients was found in both groups, which was more expressed in elderly patients. The dynamics of blood markers in patients of the 1st and 2nd groups after the treatment showed a decreasing the level of inflammatory and dystrophic processes in the knee joints. In the 1st group 1 month after intra-articular injection, the content of glycoproteins and chondroitin sulfates (CS) decreased, did not reach the normative values. In subgroup II-c the level of glycoproteins and CS decreased, the concentration of interleukin-6 (IL-6) remained elevated comparing to subgroups II-a and II-b. Conclusions: it was found that patients with stage II knee osteoarthritis had increased CS and lower levels of hyaluronic acid in their synovial fluid. Markers of inflammation (glycoproteins and IL-6) and destruction markers (CS) in the blood serum changed unidirectionally and indicated a decreasing in the activity of inflammatory process after the injection of hyaluronic acid gel, which was clinically confirmed 1 month after intra-articular injection. In this case, the results after the reexamination of blood biomarkers of patients aged 40–50 years indicated a higher efficiency of hyaluronic acid gel injection with a concentration of 10 and 12 mg/ml, aged 50–70 years — a concentration of 20 mg/ml was confirmed by a decrease in the level of glycoproteins, CS and IL-6 in the blood of patients.

Relation of Endometrial Hyaluronic Acid with Female Infertility in Women Undergoing Ovarian Stimulation Protocol

A comparative prospective study was conducted in the High Institute of Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University, from September 2019 to March 2020. The aim of the study was to evaluate the level of hyaluronic acid (HA) in endometrial fluid at day of trigger in infertile women whom undergoing ovarian stimulation protocol. The study included a total of 60 women who were selected from those attended the High Institute of Infertility Diagnosis and Assisted Reproductive Technologies. All couples subjected to a full history taking, complete general examination, complete gynecologic examination and infertility workup including: husband’s semen analysis, hormonal analysis, uterine cavity and tubal patency was performed. Different ovulation induction programs were used evaluation of endometrial thickness, pattern, size and number of mature follicles was performed by transvaginal sonography in the 2nd of menstrual cycle and cycle day 11-14 before HCG injection. In this study basal hormonal analysis were performed to the female in the 2nd or 3rd day of the menstrual cycle and study included aspiration of endometrial fluid for determination of hyaluronic acid level by ELISA technique. The study showed that mean of hyaluronic acid was elevated significantly in women received letrozole plus FSH (158.7±19.7 pg/ ml) followed by women received Clomid plus FSH (160.8±13.5 pg/ml) and the lowest mean was recorded in the control group (146.1±26.2 pg/ml). The study showedpositive correlation betweenhyaluronic acid and LH at day of triggering of ovulation in women received Clomid plus FSH (group B). The study showed positive correlation betweenhyaluronic acid and progesterone at day of triggering of ovulation in women received letrozole plus FSH. The study showed positive correlation betweenhyaluronic acid and number of follicles in women after receiving letrozole plus FSH. The study revealed that HA mean was elevated significantly (P&lt; 0.05) in women who became pregnant after stimulation protocol compared with women who failed to be pregnant (164.2 vs. 152.1 pg/ml) as shown in Figure 4). The study also revealed that HA was significantly elevated in women who became pregnant after receiving letrozole + FSH (164.2 pg/ ml) as compared with non pregnant women (152.1 pg/ml) while HA level was non significantly related with pregnancy occurrence after administration of Clomid+FSH. It was concluded that, hyaluronic acid was elevated significantly in women received ovulation simulation drugs and letrozolewas superior to clomiphene as a treatment foranovulatory infertility in women

Nucleoside analogs assisted with Chinese compound prescription in treating hepatic fibrosis of chronic hepatitis B patients: A protocol of systematic review and meta-analysis.

Background:Chronic hepatitis B is often complicated with different degrees of hepatic fibrosis, which affects the quality of life. Nucleoside analogs are recommended by almost all guidelines in the world for the treatment of chronic hepatitis B. At present, there is no specific and effective chemical and biological agents for hepatic fibrosis. In China, Chinese compound prescription combined with nucleoside analogs have been used to treat hepatic fibrosis of chronic hepatitis B patients in more and more cases, and good results have been achieved. Several Chinese compound prescriptions that have been made into proprietary Chinese medicine for the convenience of use. This article aims to systematically evaluate the efficacy and safety of Chinese medicine compounds assisting nucleoside analogs in the treatment of hepatic fibrosis in chronic hepatitis B patients.Method:The following databases will be searched from their inception to September 2019: PubMed, EMBASE, EBSCOhost, The Cochrane Library, China National Knowledge Infrastructure (CNKI), Chinese Biomedical literature Database (CBM), VIP Database, Wanfang Database. Languages are limited to Chinese and English. The study includes randomized controlled trials using Chinese compound prescription combined with entecavir and Chinese compound prescription combined with tenofovir disoproxil fumarate to treat hepatic fibrosis of chronic hepatitis B patients. The primary outcomes including effective rate and biochemical parameters (levels of hyaluronic acid, laminin, pre-type-III collagen and type IV collagen will be tested. Additional outcomes include liver function indexes (levels of alanine aminotransferase, aspartate aminotransferase, total bilirubin) and levels of hepatitis B virus DNA. Stata14.0 software will be used for meta-analysis.Result:The efficacy and safety of Chinese compound prescriptions assisting nucleoside analogs for hepatic fibrosis of chronic hepatitis B patients will be assessed from the effective rate, biochemical parameters, liver function indexes, and levels of hepatitis B virus DNA.Conclusion:The conclusion of this study will be used to evaluate the efficacy and safety of Chinese compound prescriptions assisting nucleoside analogs in the treatment of hepatic fibrosis of chronic hepatitis B patients, as well as the adjuvant effectiveness of Chinese compound prescriptions in combined therapy.PROSPERO registration number:CRD42020156859.

Normothermic Ex Vivo Liver Perfusion Prevents Intrahepatic Platelet Sequestration After Liver Transplantation

The detrimental role of platelets in sinusoidal endothelial cell (SEC) injury during liver transplantation (LT) has been previously addressed after static cold storage (SCS), however, it is currently unknown after normothermic ex vivo liver perfusion (NEVLP). Pig LT was performed with livers from heart-beating donors or donation after circulatory death (DCD) donors subjected to SCS or NEVLP (n = 5/group). All pigs except for 1 (DCD-SCS-group) survived 4 days. The heart-beating donor- and DCD-NEVLP-groups showed significantly lower aspartate transaminase-levels compared with the SCS-groups 3 hours post-LT (P = 0.006), on postoperative day (POD) 2 (P = 0.005), POD3 (P = 0.007), and on POD4 (P = 0.012). Post-LT total platelet count recovered faster in the NEVLP than in the SCS-groups at 12 hours (P = 0.023) and 24 hours (P = 0.0038). Intrahepatic sequestration of platelets was significantly higher in the SCS-groups 3 hours postreperfusion and correlated with severity of SEC injury. In both SCS-groups, levels of tumor growth factor-β were higher 3 hours post-LT, on POD1 and on POD3. Moreover, platelet factor 4 levels and platelet-derived extracellular vesicles were increased in the SCS-groups. Hyaluronic acid levels were significantly higher in the SCS-groups, indicating a higher grade of endothelial cell dysfunction. Platelet inhibition achieved by pretreatment with clopidogrel (n = 3) partly reversed the detrimental effects on SEC injury and therefore provided further evidence of the important role of platelets in ischemia/reperfusion injury and SEC injury. Normothermic perfusion of liver grafts before transplantation effectively reduced platelet aggregation and SEC injury, which translated into an improved posttransplant organ function.

1804-P: HM15211, a Novel Long-Acting GLP-1/GIP/Glucagon Triple Agonist, Exhibits Anti-inflammatory and Fibrotic Effects in AMLN/TAA–Induced Liver Inflammation and Fibrosis Mice

Nonalcoholic steatohepatitis (NASH), a severe form of NAFLD, is characterized by hepatic steatosis, inflammation, and fibrosis. Despite increased prevalence and related mortality, no approved medication is available, leading to a significant burden for public health. Although advances in targeting hepatic steatosis demonstrate meaningful benefits, their overall impact in inflammation and fibrosis remains elusive. Hence, recent approaches directly modulating inflammation and fibrosis still showed marginal efficacy. Thus, to provide novel option for NASH treatment, HM15211, a novel long-acting GLP-1/GIP/Glucagon triple agonist, is developed. Because of the benefits of GLP-1, GIP, and Glucagon in liver inflammation and fibrosis in addition to steatosis, HM15211 might confer more effective treatment in NASH. Previously, HM1521 treatment led to potent liver fat reduction along with weight loss in DIO mice. Here, we investigated the potential effect of HM15211 in animal model of liver inflammation and fibrosis. To induce liver inflammation and fibrosis, mice fed with AMLN diet were concomitantly treated with thioacetamide (AMLN/TAA mice) for 16 weeks, and HM15211 was administered during last 8 weeks. Of note, HM15211 treatment reduced the hepatic marker expression for inflammation (-92.8, -34.7 and -34.9% vs. vehicle (Veh) for F4/80, MCP-1 and IL-6) and fibrosis (-53.9, -41.4 and -51.9 % vs. Veh for α-SMA, TIMP-1 and collagen1a1). Similar reduction in blood level of TIMP-1, PIIINP and hyaluronic acid (-49.3, -48.0 and -49.1% vs. Veh) was observed. Strikingly, HM15211 significantly reduced both hepatic hydroxyproline (-53.1 % vs. Veh) and sirius red positive area (-70.6% vs. Veh). Therefore, in addition to steatosis improvement, HM15211 may provide anti-inflammatory and -fibrotic effect. Human studies are ongoing to assess the clinical relevance of these findings. Disclosure J. Kim: Employee; Self; Hanmi Pharmaceutical. J. Lee: Employee; Self; Hanmi Pharmaceutical. E. Park: Employee; Self; Hanmi Pharmaceutical. J. Lee: Employee; Self; Hanmi Pharmaceutical. S. Bae: Employee; Self; Hanmi Pharmaceutical. D. Kim: Board Member; Self; Hanmi Pharmaceutical. I. Choi: Board Member; Self; Hanmi Pharmaceutical.

Hyaluronic acid predicts poor prognosis in breast cancer patients: A protocol for systematic review and meta analysis.

Hyaluronic acid (HA) may be a novel prognostic biomarker of breast cancer. However, the available evidence is controversial. Therefore, we performed a meta-analysis to determine the prognostic role of HA in breast cancer. The data were extracted from seven articles by searching the databases of PubMed, EMBASE, Web of Science, the Chinese National Knowledge Infrastructure and Wanfang data for the prognostic role of HA in breast cancer. In reference to survival outcomes, the pooled hazard ratios (HRs) of HA were calculated given a 95% confidence interval (CI). A total of seven articles were included in our study involving 2664 cases. The result of meta-analysis showed that a high HA level predicts poor overall survival (OS) (HR = 1.86, 95% CI: 1.28-2.71, P = .001) and shortened disease-free or recurrence-free survival or progression free survival (DFS/RFS/PFS) (HR = 1.63, 95% CI: 1.14-2.33, P = .007) in breast cancer patients. Moreover, a high HA level in stroma (HR = 1.63, 95% CI: 1.06-2.51, P = .025) and plasma (HR = 3.26, 95% CI: 2.25-4.73, P < .001) significantly predicted poor OS. Besides, a tendency shows that HA was significantly correlated with lymph node metastasis (HR = 1.55, 95% CI: 0.96-2.49, P = .070) and tumor grade (HR = 2.10, 95% CI: 0.89-4.96, P = .089) on the clinical characteristics of patients. These results suggested that HA has a potential to be prognostic biomarker in breast cancer patients, especially location in stroma and plasma.

Dose Dependent Effects of Whole Blueberry on Cartilage Health and Pain in a Monosodium Iodoacetate (MIA) Induced Rat Model of Osteoarthritis

Abstract Objectives The purpose of this study was to examine the dose-dependent effects of whole blueberry in a monosodium iodoacetate (MIA) induced rat model of Osteoarthritis (OA). There is no known cure for OA and the present pharmacological treatment options are limited and associated with adverse side effects. We hypothesize that the addition of whole blueberry into the diets of rats with MIA induced OA will reduce pain behavior and improve cartilage health. Methods A total of forty, 45-day-old female CD rats were used for this study. Thirty rats were injected with MIA to induce joint destruction associated with OA. Ten rats served as control without induction of joint destruction. The MIA induced rats were randomized into three groups consisting of 10 animals. All groups were fed a casein-based diet with two of the MIA induced groups receiving an addition of whole blueberry powder at 5% and 10%, respectively. All groups were fed the respective diets for 46 days. The animals were weighted weekly throughout the study period and food intake monitored and recorded. Fasted blood specimens and other tissues of interest were collected after euthanasia for analysis. Mechanical allodynia was assessed at four time points throughout the study to evaluate changes in pain behaviors. Results There were no difference in body weight between the four groups at baseline. After 46 days of feeding there was no significant change in body weight in either of the blueberry fed groups in comparison to the MIA rats. Plasma hyaluronic acid levels were higher in the MIA group in comparison to the blueberry treated groups. We observed a dose dependent decrease in hyaluronic acid levels with blueberry treatment, respectively. At baseline, the MIA group exhibited significant (P ≤ 0.05) mechanical allodynia compared to the control group. Mechanical allodynia was significantly (P ≤ 0.05) reduced after 40 days in the 10% whole blueberry treated group. Conclusions Incorporation of whole blueberry into the diets of rats with MIA-induced OA reduced hyaluronic acid concentration and pain sensitivity. These results suggest that whole blueberry may be an effective therapeutic alternative for improving joint health and alleviating pain associated with OA. Funding Sources Research Enhancement Program, Texas Woman's University.