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Introduction Hepatitis B-related compensated liver cirrhosis is related to a higher risk of hepatocellular carcinoma, and antiviral therapy is the preferred method. As the pathological mechanisms of liver fibrosis are complex, drugs developed for a single target are difficult to be effective in clinical practice, so there are no chemical drugs or biological drugs with clear efficacy available for clinical application at present. Traditional Chinese medicine is a kind of medical science that has been gradually formed during thousands of years and continuously enriched by the people of all ethnic groups in China. Traditional Chinese medicine shows curative effects in the treatment of liver diseases, especially in the field of liver fibrosis prevention and treatment. This study aims to test the integrative medicine (Chinese medicine plus antiviral therapy) effective on lowing hepatocellular carcinoma risk among patients with hepatitis-related compensated liver cirrhosis. Methods and analysis This is a multi-center randomized controlled trial, and a total of 5 hospitals and 802 patients will be involved in. All the subjects are randomly allocated to the YinQiSanHuang Jiedu decoction (YQSHD) group (n = 401) or the placebo group (n = 401). The YQSHD group receives YQSHD granule with entecavir (ETV), and the placebo group receives YQSHD placebo with ETV. The treatment period will last for 52 weeks, and the follow-up period for 52 ± 2 weeks. The primary outcome measure is the annual incidence of HCC. Outcomes will be assessed at baseline and after treatment. The objective of this trial is “the integrative of YQSHD with ETV reduce the annual incidence of HCC to 1%.” Ethics and dissemination The protocol has been approved by the Medical Ethics Committee of Guang’anmen Hospital, China (No.2019-006-KY), and the other centers in the trial will not begin recruiting until the local ethical approval has been obtained. Trial final results will be disseminated via publication. Trial registration Chinese Clinical Trial Registry ChiCTR1900021532. Registered on February 26, 2019

Alzheimer's disease (AD) is a chronic neurodegenerative disease characterized by gradual decline of cognitive function, which is also one of the main causes of cognitive and memory impairment in the elderly. The delicate balance between the pro-inflammatory and anti-inflammatory properties of microglia determines its role in the occurrence and development of AD. Κ-carrageenan oligosaccharide (KOS) was obtained by enzymatic hydrolysis of carrageenan from Marine red algae, which can inhibit the over activation of microglia. In this study, APP/PS1 transgenic mice were used to study the effect of KOS on the symptoms of AD mice by detecting the ability of learning and memory, the activation of microglia cells, the expression of related proteins in the brain, and the deposition of APP and Aβ1−42. The expression of inflammatory cytokines and inflammation-related proteins in microglia cells were detected in vitro, and the effects of KOS on over activation of microglia cells were determined. In addition, microglia and nerve cells were cultured in transwell co-culture system to study the effect of microglia cells treated with KOS on the activity of nerve cells. The results showed that KOS could inhibit clinical symptoms and inflammatory responses by microglia of AD mice. The apoptosis of nerve cells caused by the inflammatory response of microglia was also decreased. KOS could improve the spatial learning and memory ability of AD mice and inhibit over activation of microglia, which has a promising application in the prevention and treatment of AD.

Microglia are the main effector cells of immune response in central nervous system and are important targets for disease prevention and treatment. Κ-carrageenan Oligosaccharide (KOS), obtained by enzymatic hydrolysis from carrageenan of marine red algae, can inhibit the release of inflammatory factors from the over-activated microglia. The mechanism of microglia autophagy induced by KOS and its relationship with inflammation were studied to explore the development prospect of KOS in the research and treatment of inflammatory related diseases. The effect of KOS on inducing autophagy was detected by the secretion of cytokines by lipopolysaccharide (LPS)-activated microglia, respectively. The protein expression of autophagy-related signaling pathways were detected by Western Blot. The results showed that KOS could significantly protect the microglia from over-activated inflammatory by inducing the autophagy and inhibiting the release of inflammatory cytokines. And KOS could reduce the expression of the protein that related to the AMPK/ULK1 pathways in microglia, so as to regulate the autophagy pathway, and inhibit the inflammatory response of over-activated microglia. The study on the effect of KOS on microglia autophagy and excessive inflammatory response will provide a theoretical basis for further studies on the inhibition of nerve injury by regulating microglia autophagy and inflammatory response.

Introduction: Hepatitis B-related compensated liver cirrhosis is related to higher risk of hepatocellular carcinoma, anti-viral therapy is the preferred method. As the pathological mechanisms of liver fibrosis are complex, drugs developed for a single target are difficult to be effective in clinical practice, so there are no chemical drugs or biological drugs with clear efficacy available for clinical application at present. Traditional Chinese medicine is a kind of medical science that has been gradually formed during thousands of years and continuously enriched by the people of all ethnic groups in China. Traditional chinese medicine shows curative effects in the treatment of liver diseases, especially in the field of liver fibrosis prevention and treatment. This study aim to test the integrative medicine (chinese medicine plus anti-riral therapy) effective on lowing hepatocellular carcinoma risk among patients with hepatitis B-related compensated liver cirrhosis.Methods and Analysis: This is a multicentre randomised controlled trial, total 5 hospitals and 802 patients will involved in. All the subjects are randomly allocated to the YinQiSanHuang Jiedu decoction(YQSHD) group (n=401) or the placebo group (n=401). The YQSHD group receives YQSHD granule with Entecavir(ETV), the placebo group receives YQSHD placebo with ETV. Treatment period will last for 52 weeks, and follow-up period for 52±2 weeks. The primary outcome measure is the annual incidence of HCC. Outcomes will be assessed at baseline and after treatment. Objective of this trial is “the integrative of YQSHD with ETV reduce the annual incidence of HCC to 1%”.Ethics and dissemination:The protocol has been approved by the Medical Ethics Committee of Guang’anmen Hospital, China (No.2019-006-KY), and the other centres in the trial will not begin recruiting until local ethical approval has been obtained.Trial final results will be disseminated via publication. Trial registration: ChiCTR1900021532, this protocol was registered in the Chinese Clinical Trial Registry (URL: http://www.chictr.org.cn/searchproj.aspx) on February 26th, 2019.

Microglial inflammation plays an essential role in neurodegenerative disease. Our previous studies had shown that κ-carrageenan oligosaccharides (KOS) could inhibit the excessive activation of microglia that induced by LPS, while the interrelated mechanisms were still indistinct. Therefore, we detected the inflammatory signaling pathway on LPS-activated microglia that pretreat by different content of KOS to reveal the mechanism on KOS's inhibition of microglia inflammatory response. ELISA was used to detect the effects of KOS on the secretion of interleukin-1 (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and prostaglandin E2 (PG-2) by LPS-activated microglia, respectively. The production of reactive oxygen species (ROS) and nitric oxide (NO) in microglia cells was detected by flow cytometry, and the protein expression of immunoinflammation-related signaling pathways were detected by Western Blot. The results showed that KOS could significantly protected the microglia from the over-activated inflammatory by inhibiting the release of inflammatory cytokines and the oxidative stress response. And KOS could reduce the expression of the protein that related to the TLR4/NF-κB and p38/JNK MAPKs pathways activated by LPS in microglia. However, there may be no specific target of KOS in cells. Therefore, KOS, a natural algal source oligosaccharide, has immunomodulatory effects and can be used as a potential intervention therapy for inflammatory related neurodegenerative diseases.

In order to construct a suitable scaffold for corneal cell culture and transplantation in vitro, various chitosan-hyaluronic acid blended membranes were prepared, and the properties of the blended membranes were studied. Rabbit keratocytes were seeded onto the blended membrane surfaces and the effects of the blended membranes on cell attachment and metabolism were investigated. The results showed that chitosan and hyaluronic acid (HA) were compatible in blended membranes. HA improved the homogeneity, crystallization and transparency, and decreased the tensile strength and hydrophilicity of blended membranes within the blending ratio of 1:0.05. Under the blending ratio of 1:0.02, HA reduced the harm that the chitosan membrane caused the cells and improved the biocompatibility between cells and membranes. Rabbit keratocytes grew and formed a confluent monolayer on the chitosan-hyaluronic acid blended membrane within a blending ratio of 1:0.02. All the results indicated that chitosan and HA were compatible within the blending ratio of 1:0.02, and the blended membranes had homogeneous structure, suitable physicochemical properties and biocompatibility with cells, this implied that the blended membranes could be used as a scaffold for corneal cell culture in vitro and as a potential carrier for corneal endothelial cell transplantation.

A carrageenan-degrading marine Cellulophaga lytica strain N5-2 was isolated from the sediment of carrageenan production base. A κ-carrageenase (EC 3.2.1.83) with high activity was purified to electrophoretic homogeneity from the culture supernatant by a procedure of ammonium sulfate precipitation, dialyzing and gel filtration on SephadexG-200 and SephadexG-75. The purified enzyme was verified as a single protein on SDS-PAGE, and whose molecular weight was 40.8 kDa. The κ-carrageenase yielded a high activity of 1170 U/mg protein. For κ-carrageenase activity, the optimum temperature and pH were 35 °C and pH 7.0, respectively. The enzyme was stable at 40 °C for at least 2.5 h. The enzyme against κ-carrageenan gave a Km value of 1.647 mg/mL and a Vmax value of 8.7 μmol/min/mg when the reaction was carried out at 35 °C and pH 7.0. The degradation products of the k-carrageenase were analyzed by thin layer chromatography (TLC), high performance liquid chromatography (HPLC), electrospray ionization time-of-flight mass spectroscopy (ESI-TOF-MS) and 13C-NMR spectroscopy, and the results indicated that the enzyme was specific of the β-1,4 linkage and hydrolyzed κ-carrageenan into κ-neocarraoctaose-sulfate and κ-neocarrahexaose-sulfate first, and then broke κ-neocarraoctaose-sulfate into κ-neocarrabiose-sulfate and κ-neocarrahexaose-sulfate.

In order to construct a suitable scaffold for corneal cell culture and transplantation in vitro, different chitosan-hyaluronic acid blended membranes were prepared, and the properties of the blended membranes were studied. Rabbit keratocytes were seeded onto the blended membrane surfaces and the effects of the blended membranes on cell attachment and metabolism were investigated. The results showed that chitosan and hyaluronic acid (HA) were compatible in blended membranes. HA improved the homogeneousness, crystallization and transparency, and decreased the tensile strength and hydrophilism of blended membranes within the blending ratio of 1∶0.05. Under the blending ratio of 1∶0.02, HA reduced the harm that the chitosan membrane caused the cells and improved the biocompatibility between cells and membranes. All the results indicated that chitosan and HA were compatible within the blending ratio of 1∶0.02, and the blended membranes had homogeneous structure, suitable physicochemical properties and biocompatibility with cells, which implied that the blended membranes could be used as a scaffold for corneal cell culture in vitro and as a potential carrier for corneal endothelial cell transplantation.

In order to construct a suitable scaffold for corneal cells culture and transplantation in vitro, different chitosan- hyaluronic acid blended membranes were prepared and the compatibility of chitosan and hyaluronic acid in blended membranes was studied. Rabbit keratocytes were seeded onto the blended membranes surface and the effects of the blended membranes on cells attachment and metabolism were investigated. The results showed that chitosan and hyaluronic acid had good compatibility in blended membranes within a certain blended ratio. Within the blending ratio of 1:0.05, hyaluronic acid could reduce the damage of chitosan membrane to cells and improve the adhersion and proliferation of cells on blended membranes. So chitosan and hyaluronic acid had good compatibility within a certain blended ratio. Hyaluronic acid could improve the biocompatibility between cells and membranes. The blended membrane had potential to be used as a scaffold for corneal cells culture in vitro and a carrier for corneal cells transplantation. Keywords-chitosan; hyaluronic acid; blended membrane; keratocytes