Applications of Hyaluronic Acid (HA) in Dental Implant Treatment: A Systematic Review

@#Introduction: Hyaluronic acid (HA) has a long history and is widely used in cosmetics, medicine, and dermatology. This molecule is still considered relatively new in the field of dentistry. This study aimed to assess the application of HA in dental implant treatment. Method: Search in the multiple indexed databases such as Pubmed, COCHRANE, and Scopus was conducted up until August 2022 using the keywords “hyaluronic acid”, “hyaluronan,” and “dental implant.” Results: The literature search identified 816 articles, and 17 were selected in this study. Three domains of use of HA in dental implant treatment were identified: surface modification of implant surface, treatment after insertion of a dental implant, and bone graft/membrane material. There are eight randomized control trials and nine non-randomized control trials included in this study. Only six studies showed statistically significant results with HA groups. Conclusion: Overall, there are positive findings on the application of HA in dental implant treatment, showing it can be used in dental implantology, with multiple categories of uses.

Monitoring non-small cell lung cancer progression and treatment response through hyaluronic acid in sputum

We evaluated whether hyaluronan (HA) levels in the sputum could be used as a noninvasive tool to predict progressive disease and treatment response, as detected in a computed tomography scan in non-small cell lung cancer (NSCLC) patients. Sputum samples were collected from 84 patients with histological confirmation of NSCLC, 33 of which were in early-stage and 51 in advanced-stage disease. Patients received systemic chemotherapy (CT) after surgery (n=36), combined CT and immunotherapy (IO) (n=15), or targeted therapy for driver mutation and disease relapse (N=4). The primary end-point was to compare sputum HA levels in two different concentrations of hypertonic saline solution with overall survival (OS) and the secondary and exploratory end-points were radiologic responses to treatment and patient outcome. Higher concentrations of HA in the sputum were significantly associated to factors related to tumor stage, phenotype, response to treatment, and outcome. In the early stage, patients with lower sputum HA levels before treatment achieved a complete tumor response after systemic CT with better progression-free survival (PFS) than those with high HA levels. We also examined the importance of the sputum HA concentration and tumor response in the 51 patients who developed metastatic disease and received CT+IO. Patients with low levels of sputum HA showed a complete tumor response in the computed tomography scan and stable disease after CT+IO treatment, as well as a better PFS than those receiving CT alone. HA levels in sputum of NSCLC patients may serve as a candidate biomarker to detect progressive disease and monitor treatment response in computed tomography scans.

Bioactive hyaluronic acid fragments inhibit lipopolysaccharide-induced inflammatory responses via the Toll-like receptor 4 signaling pathway / 医学前沿

The high- and the low-molecular weight hyaluronic acids (HMW-HA and LMW-HA, respectively) showed different biological activities in inflammation. However, the role of LMW-HA in inflammatory response is controversial. In this study, we aimed to investigate the effect of bioactive hyaluronan (B-HA) on lipopolysaccharide (LPS)-induced inflammatory responses in human macrophages and mice. B-HA was produced from HA treated with glycosylated recombinant human hyaluronidase PH20. Human THP-1 cells were induced to differentiate into macrophages. THP-1-derived macrophages were treated with B-HA, LPS, or B-HA + LPS. The mRNA expression and the production of inflammatory cytokines were determined using quantitative real-time PCR and enzyme-linked immunosorbent assay. The phosphorylation levels of proteins in the nuclear factor-κB (NF-κB), mitogen-activated protein kinase (MAPK), and IRF-3 signaling pathways were measured using Western blot. The in vivo efficacy of B-HA was assessed in a mouse model of LPS-induced inflammation. Results showed that B-HA inhibited the expression of TNF-α, IL-6, IL-1, and IFN-β, and enhanced the expression of the antiinflammatory cytokine IL-10 in LPS-induced inflammatory responses in THP-1-derived macrophages and in vivo. B-HA significantly suppressed the phosphorylation of the TLR4 signaling pathway proteins p65, IKKα/β, IκBα, JNK1/2, ERK1/2, p38, and IRF-3. In conclusion, our results demonstrated that the B-HA attenuated the LPS-stimulated inflammatory response by inhibiting the activation of the TLR4 signaling pathway. B-HA could be a potential anti-inflammatory drug in the treatment of inflammatory disease.

Association of htra1 and hapln1 gene polymorphism with intervertebral disc degeneration / 中国组织工程研究

BACKGROUND: Studies have found that single nucleotide polymorphism genotypes in the HTRA1 gene promoter region are associated with intervertebral disc degeneration, while HAPLN1 is associated with osteoarthritis caused by intervertebral disc degeneration. OBJECTIVE: To explore the role of human secretory serine protease HTRA1 and the key group of extracellular matrix HAPLN1 in the pathogenesis of intervertebral disc degeneration. METHODS: This study included 498 postmenopausal female subjects who underwent a physical examination at Dingzhou People’s Hospital from April 2015 to December 2018. TaqMan PCR was used to detect HTRA1 gene promoter rs11200638 single nucleotide polymorphism and HAPLN1 gene 5' flanking rs975563, intron 1 rs10942332, intron 2 rs179851 and intron 4 rs4703570 single nucleotide polymorphism in 498 postmenopausal Chinese women. The correlation between the HTRA1orHAPLN1 gene polymorphisms and the radiographic features of spinal disc degeneration was analyzed. The trial has been approved by the Ethics Committee of Dingzhou People’s Hospital. RESULTS AND CONCLUSION: Among the 498 subjects with the HTRA1 gene rs11200638 single nucleotide polymorphism, 178 were GG homozygotes, 222 were GA heterozygotes, and 98 were AA homozygotes. We compared the parameters of intervertebral disc degeneration in subjects with at least one G allele (GG+GA, n=400) and without G allele (AA, n=98). In HTRA1 gene rs11200638 single nucleotide polymorphism, the score on intervertebral space stenosis in the subjects with GG+GA allele genome was lower than that in the subjects with AA allele (P < 0.001). With the increase of the score on intervertebral space stenosis, the proportion of the subjects with AA alleles increased (P ≤ 0.001). Among the 498 subjects with single nucleotide polymorphisms of the HAPLN1 gene, 137 were homozygous for TT, 230 were heterozygous for CT, and 131 were homozygous for CC. Intervertebral disc degeneration parameters of CC+TT allele (n=361) and TT allele (n=137) were compared. In the HAPLN1 gene, there was a significant difference between the CC+TT and TT alleles of the rs179851 single nucleotide polymorphism in osteophyte formation and intervertebral space stenosis (P < 0.01). Among the HAPLN1 gene rs179851 single nucleotide polymorphisms, the proportion of subjects with TT alleles and intervertebral space stenosis ≥ 6 points increased (P < 0.05). With an increase in osteophyte formation score, the proportion of subjects with TT allele increased (P < 0.001). These results reveal that HTRA1 and HAPLN1 genetic variations at specific genetic loci are associated with intervertebral disc degeneration.

Cloning and expression of hyaluronan synthase (hasA) in recombinant Escherichia coli BL21 and its hyaluronic acid production in shake flask culture

Aims@#Hyaluronic acid (HA) is a high molecular weight polymer and a major component of mucoid capsule in bacteria and extracellular matrix (ECM) of vertebrate tissue. Due to its unique characteristics, HA is used extensively in medical and cosmetic field. However, because of the exotoxins production from animal tissues extraction and Streptococcus zooepidemicus, HA production by recombinant microorganisms has gained interest. The present study was aimed at cloning of hasA gene in Escherichia coli and optimization of the medium components for HA production. @*Methodology and results@#A fragment of an approximate size of 1.5 kb that encodes the hyaluronan synthase (hasA) gene from S. zooepidemicus ATCC 39920 was amplified by PCR using hasA-specific primers. The hasA gene was ligated into the bacterial expression vector pLbADH and transformed into the expression host, Escherichia coli BL21 strain. Then, genetically engineered E. coli strain BL21 was used for the production of HA by fermentation using different glucose concentration (10-50 g/L) and different IPTG concentration (0.1, 0.5 and 1.0 mM) in shake flask culture. Amongst varying glucose concentrations, results showed that 50 g/L glucose with nutrient rich media containing nitrogen source was able to produce the highest HA concentration (0.115 ± 0.002 g/L). With addition of 1.0 mM IPTG, HA production reached a peak 0.532 ± 0.026 g/L which is around fivefold higher compared to without IPTG. @*Conclusion, significance and impact of study@#The hasA gene was cloned from S. zooepidemicus and successfully expressed in recombinant E. coli BL21 cells. This low molecular weight HA is gaining more importance in medical and cosmetic application due to possess pronounced free radical scavenging and antioxidant activities.

Effect of a new cross-linked hyaluronan gel on the staple line after sleeve gastrectomy in a rat model

Abstract Purpose: To evaluate the effect of a new cross-linked hyaluronan (NCHA) gel on healing of the staple line in an experimental sleeve gastrectomy. Methods: Eighteen rats were randomly divided into three groups. The control group (n = 6) received no medication. In the saline group (n = 6) and NCHA gel group (n = 6), saline and NCHA gel were respectively administered onto the staple line and intraperitoneally into the abdominal cavity after the standard stapling procedure. Results: The fibroblast activity and collagen deposition were significantly higher in the NCHA gel group than in the control group (p = 0.00, p = 0.017) and saline group (p = 0.004, p = 0.015). The tissue hydroxyproline protein level was significantly higher in the NCHA gel group than in the control group (p = 0.041). Adhesion formation was significantly lower in the NCHA gel group than in the control and saline groups (p = 0.015, p = 0.041). Conclusions: New cross-linked hyaluronan gel could be an effective approach to improve staple line wound healing and prevent potential leakage after sleeve gastrectomy. Moreover, NCHA gel helps to prevent adhesion formation without compromising healing of the staple line.

TGF-β1 upregulates the expression of hyaluronan synthase 2 and hyaluronan synthesis in culture models of equine articular chondrocytes

We investigated the effect of transforming growth factor beta 1 (TGF-β1) on equine hyaluronan synthase 2 (HAS2) gene expression and hyaluronan (HA) synthesis in culture models of articular chondrocytes. Equine chondrocytes were treated with TGF-β1 at different concentrations and times in monolayer cultures. In three-dimensional cultures, chondrocyte-seeded gelatin scaffolds were cultured in chondrogenic media containing 10 ng/mL of TGF-β1. The amounts of HA in conditioned media and in scaffolds were determined by enzyme-linked immunosorbent assays. HAS2 mRNA expression was analyzed by semi-quantitative reverse transcription polymerase chain reaction. The uronic acid content and DNA content of the scaffolds were measured by using colorimetric and Hoechst 33258 assays, respectively. Cell proliferation was evaluated by using the alamarBlue assay. Scanning electron microscopy (SEM), histology, and immunohistochemistry were used for microscopic analysis of the samples. The upregulation of HAS2 mRNA levels by TGF-β1 stimulation was dose and time dependent. TGF-β1 was shown to enhance HA and uronic acid content in the scaffolds. Cell proliferation and DNA content were significantly lower in TGF-β1 treatments. SEM and histological results revealed the formation of a cartilaginous-like extracellular matrix in the TGF-β1-treated scaffolds. Together, our results suggest that TGF-β1 has a stimulatory effect on equine chondrocytes, enhancing HA synthesis and promoting cartilage matrix generation.

The role of human lysozyme-like protein 4 in fertilization and its enzymatic properties / 中华男科学杂志

Objective@#To elucidate the possible role of human lysozyme-like protein 4 (LYZL4) in fertilization and characterize its enzymatic properties.@*METHODS@#The localization of LYZL4 in human spermatozoa was investigated by immunofluorescence staining, the sources of LYZL4 on the sperm surface examined by RT-PCR, and the role of LYZL4 in fertilization assessed by the zona-free hamster egg penetration test. The recombinant plasmid pPIC9K-LYZL4 was constructed and its expression induced with methanol after transformed into competent Pichia pastoris GS115. The recombinant LYZL4 protein (rLYZL4) was purified from the fermentation supernatant and subsequently identified by Western blot. The hyaluronan binding ability of rLYZL4 was determined by ELISA and the muramidase activity, hyaluronidase activity, and free radical scavenging ability examined by spectrophotometric methods.@*RESULTS@#Immunodetection with a specific antiserum localized LYZL4 on the acrosomal membrane of mature spermatozoa, which was exclusively secreted from the testis and epididymis as shown by RT-PCR. Immunoneutralization of LYZL4 significantly decreased the number of human spermatozoa bound to zona-free hamster eggs in a dose-dependent manner in vitro. The recombinant protein was expressed successfully by the P. pastoris strain GS115. Purified rLYZL4 exhibited a potent hyaluronan binding ability and a strong free radical scavenging ability but no muramidase or hyaluronidase activity.@*CONCLUSIONS@#LYZL4 secreted from the testis and epididymis is localized on the acrosomal membrane of mature spermatozoa and plays a role in sperm-egg binding as well as in binding hyaluronan and scavenging free radicals, which suggests that it might be a multi-functional molecule contributive to sperm protection and sperm-egg binding.

Oral Administration of Glycine and Leucine Dipeptides Improves Skin Hydration and Elasticity in UVB-Irradiated Hairless Mice

Placenta is a special organ that contains many nutrients such as growth factors, minerals, and bioactive peptides. Dipeptides of glycine and leucine are major components of porcine placenta extracts (PPE) that has been used as an alternative of human placenta extracts. In this study, we investigated whether major peptides of PPE, Glycyl-L-Leucine (Gly-Leu), L-Leucyl-Glycine (Leu-Gly), and L-Leucyl-L-Leucine (Leu-Leu), affect skin hydration and elasticity in vitro and in vivo. We found that Gly-Leu and Leu-Gly dipeptides induced the expression of transglutaminase 1 in normal human epidermal keratinocytes (NHEKs) whereas Leu-Leu dipeptides did not. Treatment with Gly-Leu or Leu-Gly significantly increased hyaluronan (HA) synthesis in NHEKs and the upregulation of hyaluronan synthase 2 (HAS2) mRNA level was confirmed. In addition, elastase activity was inhibited in NHEKs treated with Gly-Leu or Leu-Gly dipeptides. Oral administration of Gly-Leu or Leu-Gly dipeptides increased skin hydration and elasticity in UVB-irradiated hairless mice. The significant upregulation of HA in UVB-irradiated hairless mice was observed in response to oral administration of Gly-Leu or Leu-Gly. These results suggest that the major dipeptides of porcine placenta, Gly-Leu and Leu-Gly, are potentially active ingredients for skin moisturization formulations.

Neutrophil elastase levels in the gingival crevicular fluid following hyaluronan gel application in the treatment of chronic periodontitis: A randomized split‑mouth study.

Background: Neutrophils are the predominant leukocytes in the periodontium, which prevent infection from periodontal pathogens and subsequent tissue destruction. A potentially destructive role has been elucidated, especially due to elastase enzyme. Controlling its levels might be crucial in minimizing the tissue destruction. Hyaluronan, known to inhibit the release of this enzyme from neutrophils, might be a viable option to treat chronic periodontitis. Aims: The aim of this study is to evaluate and compare the effects of 0.2% hyaluronan gel adjunctive to scaling and root planing on the levels of elastase in gingival crevicular fluid (GCF). Settings and Design: This split‑mouth study included eighty (forty experimental and forty control) sites from twenty patients representing both sexes. Materials and Methods: GCF samples were collected from all the eighty sites; simultaneously, clinical periodontal parameters were recorded. Enzyme‑linked immunosorbent assay was used to determine the levels of elastase at baseline and 6 weeks after therapy, following mechanical debridement and subsequent subgingival placement of the experimental drug. Statistical Analysis Used: With the aid of statistical software (SPSS Version 13), Student’s t‑test and Pearson’s correlation test were performed. Results: There was a mean reduction in the elastase levels from baseline to 6 weeks after therapy in the experimental group. However, the difference between the groups was not statistically significant. Conclusions: Adjunctive use of hyaluronan following mechanical debridement resulted in comparable reduction in the elastase levels, suggesting that this substance has an inhibitory effect on elastase, and subsequent tissue destruction. Further long‑term studies are mandatory to validate the results of this study.

Free radical scavenger activity of chitosan and its aminated derivative.

Antioxidants may have a positive effect on human health since they can protect the human body against deterioration by reactive oxygen species (ROS). Chitosan has many promising properties as nontoxic, biocompatible, biodegradable, antimicrobial and recently antioxidant properties. Most of the reports on the antioxidant activity of chitosan are based on the ability of amine and hydroxyl group to scavenge free radical to form stable macromolecular radical. In the current study, a new chitosan derivative with extra free amine groups was evaluated using two popular antioxidant evaluation methods (uninhibited/inhibited hyaluronan degradation and decolorization of ABTS method). The result obtained in our study show increase the activity of aminated chitosan for scavenging free radicals than chitosan itself.

Free Radical Scavenger Activity of Cinnamyl Chitosan Schiff Base.

In this study, new cinnamyl chitosan schiff base was evaluated as antioxidant material. Antioxidant activity was measured by two different popular methods (uninhibited/inhibited hyaluronan degradation and decolorization of ABTS methods). the results show decrease the hydrogen donation behavior of chitosan after coupling with cinnamaldehyde, in the other hand, ABTS method show increase the electron donation activity of cinnamyl chitosan than the chitosan itself.

The effect of hydrochloric acid stimulation and mechanical stretch on epithelial-mesenchymal transition in ;human lung epithelial cells / 中华危重病急救医学

Objective To investigate the effect of hydrochloric acid (HCl) stimulation and mechanical stretch on epithelial-mesenchymal transition (EMT) and hyaluronan (HA) production in human lung epithelial cells. Methods Human lung epithelial cell line BEAS-2B was cultured in vitro, which was divided into phosphate-buffer saline (PBS) + static group, HCl + static group, PBS + stretch group, and HCl + stretch group respectively in the logarithmic phase. The BEAS-2B cells in two stretching groups were challenged by cyclic stretch with 20% amplitude, frequency of 0.33 Hz, sine wave of the FX-5000T system for 48 hours. The morphology changes in cells before and after stretch were observed with inverted microscope. The protein expressions of epithelial markers E-cadherin and cytokeratin-8 (CK-8) as well as mesenchymal markers vimentin and α-smooth muscle actin (α-SMA) were determined by Western Blot. The secretion of HA was determined by enzyme linked immunosorbent assay (ELISA). Results ① It was shown by microscopic observation that BEAS-2B cells displayed cobblestone morphology, linked closely and cell polarity in PBS + static group, which did not change obviously after HCl stimulation alone. Given purely mechanical stretch after 48 hours, the cells morphology changed from cobblestone shape into long spindle, and increased intercellular space obviously. Double hit of HCl and stretch changed the cells morphology more significantly. ② It was shown by Western Blot that compared with the PBS + static group, HCl alone or combined with purely mechanical stretch after 48 hours, the expressions of E-cadherin and CK-8 were decreased, while those of vimentin and α-SMA were increased, and it was more pronounced in HCl + stretch group [the expression quantity (gray value) as base 1 in PBS + static group, E-cadherin: 0.16±0.08 vs. 1, CK-8: 0.10±0.03 vs. 1, vimentin: 3.35±0.38 vs. 1, α-SMA: 3.10±0.45 vs. 1, all P < 0.01]. ③ It was shown by ELISA that both HCl stimulation and stretch could induce BEAS-2B cells secreting HA as compared with PBS + static group (μg/L: 55.763±0.687, 63.005±0.493 vs. 49.876±1.867), and the production of HA increased more remarkably after double hit (μg/L: 78.220±1.085 vs. 49.876±1.867, P < 0.01). Conclusions Both HCl and mechanical stretch could induce EMT and increase HA secretion in human lung epithelial cells in vitro. Double hit of HCl stimulation and mechanical stretch induced EMT apparently, and further increased the production of HA.

Targeting hyaluronan for the treatment of pancreatic ductal adenocarcinoma

Progression of cancer is often associated with interactions between cancer cells and extracellular matrix (ECM) surrounding them. Increasing evidence has suggested that accumulation of hyaluronan (HA), a major component of ECM, provides a favorable microenvironment for cancer progression. Pancreatic ductal adenocarcinoma (PDAC) is characterized typically by a dense desmoplastic stroma with a large amount of HA, making this molecule as an attractive target for therapy. Several studies have shown efficacy of inhibitors of HA synthesis or signaling for the treatment of PDAC. Recent studies have also demonstrated substantial improvements in the effects of chemotherapy by a targeted depletion of stromal HA in PDAC using an enzymatic agent. Thus, targeting HA has been recognized as a promising therapeutic strategy to treat this highly aggressive neoplasm. In this review article, we summarize our current understanding of the role of HA in the progression of PDAC and discuss possible therapeutic approaches targeting HA.

Hyaluronidases and hyaluronan synthases expression is inversely correlated with malignancy in lung/bronchial pre-neoplastic and neoplastic lesions, affecting prognosis

We collected a series of 136 lung/bronchial and 56 matched lung parenchyma tissue samples from patients who underwent lung/bronchial biopsies and presented invasive carcinoma after lung surgery. The lung/bronchial samples included basal cell hyperplasia, squamous metaplasia, moderate dysplasia, adenomatous hyperplasia, severe dysplasia, squamous cell carcinoma and adenocarcinoma. Matched lung parenchyma tissue samples included 25 squamous cell carcinomas and 31 adenocarcinomas. Immunohistochemistry was performed to analyze for the distribution of hyaluronidase (Hyal)-1 and −3, and hyaluronan synthases (HAS)-1, −2, and −3. Hyal-1 showed significantly higher expression in basal cell hyperplasia than in moderate dysplasia (P=0.01), atypical adenomatous hyperplasia (P=0.0001), or severe dysplasia (P=0.03). Lower expression of Hyal-3 was found in atypical adenomatous hyperplasia than in basal cell hyperplasia (P=0.01) or moderate dysplasia (P=0.02). HAS-2 was significantly higher in severe dysplasia (P=0.002) and in squamous metaplasia (P=0.04) compared with basal cell hyperplasia. HAS-3 was significantly expressed in basal cell hyperplasia compared with atypical adenomatous hyperplasia (P=0.05) and severe dysplasia (P=0.02). Lower expression of HAS-3 was found in severe dysplasia compared with squamous metaplasia (P=0.01) and moderate dysplasia (P=0.01). Epithelial Hyal-1 and −3 and HAS-1, −2, and −3 expressions were significantly higher in pre-neoplastic lesions than in neoplastic lesions. Comparative Cox multivariate analysis controlled by N stage and histologic tumor type showed that patients with high HAS-3 expression in pre-neoplastic cells obtained by lung/bronchial biopsy presented a significantly higher risk of death (HR=1.19; P=0.04). We concluded that localization of Hyal and HAS in lung/bronchial pre-neoplastic and neoplastic lesions was inversely related to malignancy, which implied that visualizing these factors could be a useful diagnostic procedure for suspected lung cancer. Finalizing this conclusion will require a wider study in a randomized and prospective trial.

Soluble CD40L promotes proliferation of orbital fibroblasts and expression of hyaluronan synthase in patients with thyroid associated ophthalmopathy / 第二军医大学学报

Objective To investigate the effects of soluble CD40L (sCD40L) on proliferation of orbital fibroblasts (OFs) and the expression of three types of hyaluronan synthase (HAS) in vitro, so as to explore the role of sCD40L in the pathogenesis of thyroid-associated ophthalmopathy (TAO). Methods OFs obtained from 5 patients with TAO and 3 normal controls were primarily culutred. The effect of different concentrations of sCD40L (6.25,12.5,25,50,100 and 200 ng/mL) on proliferation of OFs of different sources were examined by MTS after 48 h exposure. OFs were also cultured with different concentrations of sCD40L (12.5, 25, 50 and 100 ng/mL) for 3,6,12 and 24 h, and then the expression levels of HAS 1-3 mRNA were determined by real-time RT-PCR. Results Treatment with sCD40L at concentrations higher than 25 ng/mL for 48 h obviously promoted the proliferation of OFs in patients with TAO (P<0.05). In contrast, treatment with sCD40L only at concentrations higher than 50 ng/mL for 48 h could promote proliferation of OFs from normal control, and the effect was comparatively weak. HAS3 mRNA expression of OFs in TAO patients was increased after exposed to sCD40L (P<0.05), and the increase was in a concentration- and time-dependent manner. Conclusion sCD40L can promote the proliferation of OFs and expression of HAS3 mRNA in patients with TAO, which implies that sCD40L plays an important role in the pathogenesis of TAO.

Effects of VEGF-C, VEGFR-3 and LYVE-1 on Lymphangiogenesis and Prognosis in Gastric Cancer / 天津医药

Objective To study the role of expressions of vascular endothelial growth factor C (VEGF-C), vascular endothelial growth factor receptor-3 (VEGFR-3) and lymphatic vessel hyaluronan receptor-1(LYVE-1) on the lymphangio-genesis and prognosis in gastric cancer. Methods The tissue microarray technology was used to detect the expressions of VEGF-C, VEGFR-3 and LYVE-1 in 125 gastric cancer specimens, 96 adjacent normal tissues and 20 benign gastric lesion samples. The lymphatic vessel density (LVD) marked by Podoplanin was detected as well. Results The positive rates of VEGF-C, VEGFR-3 and LYVE-1 in gastric cancer tissues were 62.4%, 56.0%and 58.4%, which were significantly higher than those in adjacent normal tissues (10.4%,12.5%and 9.4%) and benign gastric lesion tissues (20%, 30%and 25%, P<0.05). The LVD score was significantly higher in gastric intra-tumoral and peri-tumoral samples (2.98±0.81 and 4.22±1.09) than that in adjacent normal tissues or benign gastric lesion samples (1.82±0.63 or 0.89±0.45, P<0.01). The LVD score was significantly higher in peri-tumoral samples than that of intra-tumoral samples (P<0.01). There was a positive relationship between expression levels of VEGF-C,VEGFR-3 and LYVE-1 with LVD (P<0.05). The positive expressions of the three indexes were the risk factors of lymph node metastasis and distant metastasis (P<0.05). There was a significantly longer 5-year survival rate in patients with negative expression of the three indexes (P<0.05). Conclusion VEGF-C, VEGFR-3 and LYVE-1 proteins were positively highly expressed in gastric cancer tissues, which were risk factors affecting the progno-sis of gastric cancer. The expression levels of the three indexes can be used to predict the prognosis and lymphangiogenesis of gastric cancer.

Potential role of LYVE-1 in lymphatic metastasis of cancer / 肿瘤

This review aims to explore the mechanism of occurrence and progression of lymphatic metastasis of cancer and its role in cancer therapy. The pace of research into lymphatic system continues to accelerate with the availability of lymphatic molecular markers, such as lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1), the lymphatic receptor for hyaluronan. Recently, biological functions of LYVE-1 and its role in cancer metastasis have become the focus of worldwide attention. Clarification of the precise role of LYVE-1 in lymphatic metastasis is of great significance in exploring novel cancer treatment. This paper reviews the progress in LYVE-1 related to lymphatic metastasis of cancer. Copyright © 2013 by TUMOR.

Optimal concentration of hyaluronan and plant protein in different culture systems for in vitro maturation of bovine oocytes.

With a view to search for optimal concentration of hyaluronan (HA) and plant protein (PP) in different culture systems for in vitro maturation of bovine oocytes, cumulus-oocyte complexes (COCs) were matured in vitro in 2 culture systems (first co-cultured with granulose cells and estrus calf serum (ECS) in 2 mL volume, second without co-culture where ECS was replaced by exogenous hormones and BSA or PP in 100 µL dose under mineral oil). Seven types of media were used; 3 in first system and 4 in second system. To evaluate HA and PP effect on oocytes after in vitro culture an estimation of meiosis stage and a level of DNA fragmentation was performed by TUNEL staining. The highest meiotic maturation (84%) was observed in oocytes cultured in medium enriched with ECS in co-culture with granulose cells (1st system). The lowest meiotic maturation was noted in medium with addition of BSA (43%). The addition of HA in the medium enriched with BSA significantly increased the rate of matured oocytes (67%) and also didn’t affect the chromatin quality of individual oocytes. The addition of HA to the culture medium supplemented with a PP decreased the rate of matured oocytes to 54% but no statistical differences were noted. The results of the present study showed that HA supplementation didn’t have a detrimental impact on oocyte chromatin integrity and improved bovine oocytes’ meiotic maturation in medium supplemented only with BSA without co-culture of granulose cells.

Bioactive hyaluronan fragment (hexasaccharide) detects specific hexa-binding proteins in human breast and stomach cancer: Possible role in tumorogenesis.

Hyaluronan (HA) is a component of extracellular matrix that influences cell-proliferation, migration, development, regeneration, normal tissue remodeling, tissues undergoing malignancy and tumor cell interaction. The widespread occurrence of HA binding proteins, their involvement in tissue organization and the control of cellular behavior are well documented. The low molecular mass HA fragments can also induce a variety of biological events, including chemokine gene expression, transcription factor expression and angiogenesis. It is believed that these fragments are more potent in cellular activities than high molecular mass HA. In this study, we isolated the various fragments by gel permeation chromatography of hyaluronidase digested HA and characterized by fluoro assisted carbohydrate electrophoresis (FACE) and matrix assisted laser desorption ionization analysis (MALDI). Detection and distribution of cellular receptors in invasive tumor tissues for HA polymer and HA fragments were determined both by Western blot and histochemistry. The study demonstrated the overexpression of HA-hexa binding protein in human tumors of breast and stomach and its involvement in tumorogenesis.