Flavonoid substituted polysaccharides from Tamarix chinensis Lour. alleviate H1N1-induced acute lung injury via inhibiting complement system.

ETHNOPHARMACOLOGICAL RELEVANCE: Viral pneumonia is a highly pathogenic respiratory infectious disease associated with excessive activation of the complement system. Our previous studies found that the anticomplement polysaccharides from some medicinal plants could significantly alleviate H1N1-induced acute lung injury (H1N1-ALI). The leaves and twigs of Tamarix chinensis Lour. are traditionally used as a Chinese medicine Xiheliu for treating inflammatory disorders. Interestingly, its crude polysaccharides (MBAP90) showed potent anticomplement activity in vitro. AIM OF THE STUDY: To evaluate the therapeutic effects and possible mechanism of MBAP90 on viral pneumonia and further isolate and characterize the key active substance of MBAP90. MATERIALS AND METHODS: The protective effects of MBAP90 were evaluated by survival tests and pharmacodynamic experiments on H1N1-ALI mice. Histopathological changes, viral load, inflammatory markers, and complement deposition in lungs were analyzed by H&E staining, enzyme-linked immunosorbent assay (ELISA), and immunohistochemistry (IHC), respectively. An anticomplement homogenous polysaccharide (MBAP-3) was obtained from MBAP90 by bio-guided separation, and its structure was further characterized by methylation analysis and NMR spectroscopy. RESULTS: Oral administration of MBAP90 at a dose of 400 mg/kg significantly increased the survival rate of mice infected with the lethal H1N1 virus. In H1N1-induced ALI, mice treated with MBAP90 (200 and 400 mg/kg) could decrease the lung index, lung pathological injury, the levels of excessive proinflammatory cytokines (IL-6, TNF-α, MCP-1, IL-18, and IL-1ß), and complement levels (C3c and C5b-9). In addition, MBAP-3 was characterized as a novel homogenous polysaccharide with potent in vitro anticomplement activity (CH50: 0.126 ± 0.002 mg/mL), containing 10.51% uronic acids and 9.67% flavonoids, which were similar to the composition of MBAP90. The backbone of MBAP-3 consisted of →4)-α-D-Glcp-(1→, →3,4,6)-α-D-Glcp-(1→, and →3,4)-α-D-Glcp-(1→, with branches comprising α-L-Araf-(1→, α-D-GlcpA-(1→, →4,6)-α-D-Manp-(1→ and →4)-ß-D-Galp-(1 â†’ . Particularly, O-6 of →4)-ß-D-Galp-(1→ was conjugated with a flavonoid, myricetin. CONCLUSIONS: MBAP90 could ameliorate H1N1-ALI by inhibiting inflammation and over-activation of the complement system. These polysaccharides (MBAP90 and MBAP-3) with relative high contents of uronic acid and flavonoid substituent might be vital components of T. chinensis for treating viral pneumonia.

The Molecular Epidemiology of Prevalent Klebsiella pneumoniae Strains and Humoral Antibody Responses against Carbapenem-Resistant K. pneumoniae Infections among Pediatric Patients in Shanghai.

Carbapenem-resistant Klebsiella pneumoniae (CRKP) has caused wide dissemination among pediatric patients globally and thus has aroused public concern. Here, we investigated the clinical epidemiological characteristics of 140 nonreplicate clinical K. pneumoniae strains isolated from pediatric patients between January and December 2021. Of all isolates, 16.43% (23 of 140) were CRKP strains, which predominantly contained KPC carbapenemase. wzi sequencing demonstrated that KL47 (65.22%, 15 of 23) was the most frequent capsular type, followed by KL64 (17.39%, 4 of 23). A total of 23 CRKP strains were classified into three different O-genotypes, including OL101 (65.22%, 15 of 23), O1 (26.09%, 6 of 23), and O3 (8.7%, 2 of 23). Interestingly, KL47 strains were strongly associated with OL101, while KL64 strains were all linked with O1. Some capsule-deficient strains were identified by serological typing, phage-typing, depolymerase-typing, and uronic acid assay. In this study, compared with healthy children, higher titers of anti-capsular polysaccharides (CPS) IgG were first detected in the sera of K47 and K64 K. pneumoniae-infected children, which had the effective bactericidal activity against corresponding serotype K. pneumoniae strains. These findings will facilitate the development of novel therapeutic and vaccine strategies against K. pneumoniae infection in children. IMPORTANCE The emergence of carbapenem-resistant Klebsiella pneumoniae (CRKP) strains resistant to numerous antibiotics and the limited therapeutic options available have become an urgent health threat to the immunocompromised pediatric population. Vaccines and antibodies, especially those targeting capsular polysaccharides, may be novel and effective prevention and treatment options. Thus, it is important to understand the spread of CRKP in pediatric populations. This research presents OL101:KL47 and O1:KL64 as the predominant combinations among CRKP strains in children in Shanghai, China. The primary carbapenemase gene is KPC in CRKP strains. Additionally, this study found elevated levels of anti-CPS IgG against K47 and K64 K. pneumoniae strains in pediatric patients for the first time. The significant bactericidal activity of these anti-CPS IgGs was confirmed.

Anti-Diabetic and Angio-Protective Effect of Guluronic Acid (G2013) as a New Nonsteroidal Anti-Inflammatory Drug in the Experimental Model of Diabetes.

BACKGROUND: This study aimed to investigate the effects of guluronic acid (G2013) on blood sugar, insulin, and gene expression profile of oxLDL receptors (SR-A, CD36, LOX-1, and CD68) in the experimental model of diabetes. METHODS: 18 Sprague Dawley rats were randomly assigned to three groups of healthy control, diabetic control, and G2013 group. Diabetes was induced through intraperitoneal (IP) injection of 60 mg/kg streptozotocin. The subjects were IP treated with 25 mg/kg of G2013 per day for 28 days. The body weight, food intake, fasting blood glucose and insulin were measured. In addition, the expression of mentioned genes was investigated through quantitative real-time PCR. RESULTS: The data showed that the final weight increased significantly in the G2013-treated subjects compared to the diabetic control (p < 0.05). The results indicated that final food intake significantly reduced in the G2013-treated subjects compared to the diabetic control (p < 0.05). The study findings also suggested that the final fasting blood glucose significantly reduced in the G2013-treated group, whereas the final fasting serum insulin level significantly increased in this group compared to the diabetic control (p < 0.05). Moreover, the gene expression levels of SR-A, CD36, LOX-1, and CD68 in the G2013 group significantly reduced compared to the diabetic control (p < 0.05). CONCLUSION: This study showed that G2013, could reduce blood glucose and increase insulin levels and reduce the gene expression level of oxLDL receptors. In addition, it may probably play an important role in reducing the severity of diabetes-induced inflammatory symptoms.

Galactosaminoglycans: Medical Applications and Drawbacks.

Galactosaminoglycans (GalAGs) are sulfated glycans composed of alternating N-acetylgalactosamine and uronic acid units. Uronic acid epimerization, sulfation patterns and fucosylation are modifications observed on these molecules. GalAGs have been extensively studied and exploited because of their multiple biomedical functions. Chondroitin sulfates (CSs), the main representative family of GalAGs, have been used in alternative therapy of joint pain/inflammation and osteoarthritis. The relatively novel fucosylated chondroitin sulfate (FCS), commonly found in sea cucumbers, has been screened in multiple systems in addition to its widely studied anticoagulant action. Biomedical properties of GalAGs are directly dependent on the sugar composition, presence or lack of fucose branches, as well as sulfation patterns. Although research interest in GalAGs has increased considerably over the three last decades, perhaps motivated by the parallel progress of glycomics, serious questions concerning the effectiveness and potential side effects of GalAGs have recently been raised. Doubts have centered particularly on the beneficial functions of CS-based therapeutic supplements and the potential harmful effects of FCS as similarly observed for oversulfated chondroitin sulfate, as a contaminant of heparin. Unexpected components were also detected in CS-based pharmaceutical preparations. This review therefore aims to offer a discussion on (1) the current and potential therapeutic applications of GalAGs, including those of unique features extracted from marine sources, and (2) the potential drawbacks of this class of molecules when applied to medicine.

A randomized clinical trial for the assessment of the efficacy and safety of guluronic acid (G2013) in patients with rheumatoid arthritis.

Objective: To evaluate the safety, efficacy and tolerability of guluronic acid (G2013) in order to treat the rheumatoid arthritis patients who had inadequate response to conventional drugs. Methods: A randomized, 12-week clinical trial with two treatment arms: guluronic acid (G2013) and conventional treatment was performed. The diagnosed RA patients according to the ACR/European League against Rheumatism 2010 classification criteria, with an active disease at baseline that had inadequate response to conventional therapy were considered for the study. G2013 was administered orally twice a day with capsules of 500 mg during a period of 12 weeks and the patients were followed up for the safety and efficacy. Results: Our data showed that, the mean changes in the G2013 and control groups were -7.54 and -2.5 for tender joint count; -7.59 and -3.59 for swollen joint count; -30 and -0.9 for physician global assessment; -23.18 and -1.81 for patient global assessment; -14.45 and -1.45 for erythrocyte sedimentation rate, respectively. Improvements seen with G2013 were significantly greater than those with conventional drugs. In total, in 15.3% of G2013-treated patients and 69.2% of conventional-treated patients adverse events (AEs) occurred in this study. Conclusion: These data from routine rheumatology clinical practice highlight the effectivenessof G2013 in combination with conventional therapy with more desirable safety profile compared to the conventional-treated patients. Therefore, G2013 therapy could be an appropriate choice in order to manage the RA disease. (Clinical trial identifier: IRCT2016092813739N5).

Evaluation of the Effect of α-L-Guluronic Acid (G2013) on COX-1, COX-2 Activity and Gene Expression for Introducing this Drug as a Novel NSAID with Immunomodulatory Property.

BACKGROUND: Nonsteroidal anti-inflammatory drugs (NSAIDs) are used to treat the pathological pain and inflammation through inhibition of cyclooxygenase (COX) enzyme and disruption of the synthesis of prostaglandins (PGs). The α-L-guluronic acid (G2013) patented (PCT/EP2017/067920), as a novel NSAID with the immunomodulatory property, has been shown its positive effects in experimental models of multiple sclerosis and anti-aging. OBJECTIVE: This study was aimed to investigate the effects of G2013 on the gene expression and activity of COX-1/COX-2 enzymes in order to introduce a novel NSAID for the treatment of inflammatory diseases. METHOD: The mRNA expression levels of COX-1/COX-2 were measured by qRT-PCR. The PGE2 concentration in culture media was determined using ELISA method. RESULTS: Our results demonstrated that the low and high dose of G2013 could significantly reduce the gene expression of COX-1 and COX-2, as compared to the control treated with LPS (p < 0.05). In addition, data showed that 5, 50 and 500 mMol/ml doses of this drug can significantly the reduce activities of COX-1 and COX-2, as compared to the control treated with LPS and AA (p < 0.0001). CONCLUSION: This study revealed that G2013, as a novel NSAID with the immunomodulatory property, is able to reduce the gene expression and activity of COX-1/COX-2 enzymes. According to the findings, this agent might be categorized and introduced as a novel NSAID for the treatment of inflammatory diseases.

Angiogenesis and cardioprotection after TNFalpha-inducer-Tolpa Peat Preparation treatment in rat's hearts after experimental myocardial infarction in vivo.

The aim of the presented work was to evaluate whether short subcutaneous (s.c.) administration of TNFalpha-inducer-Tolpa Peat Preparation (TPP or TPP batch 0210) modulates the process of ischemic remodeling and spontaneous angiogenesis after experimental myocardial infarction (MI) in rats in vivo. The results obtained using three complementary and correlative methods: histological studies, Proliferating Cell Nuclear Antigen (PCNA) reaction and Lymphocytes Induced Angiogenesis (LIA) test showed a clear pro-angiogenic and cardioprotective effect of TPP administration after experimental MI. TPP batch 0210 should be considered as an angiogenesis stimulating factor and consecutively as a cardioprotective preventing development of ischemic cardiomyopathy after MI in rats. It might possibly be used as an adjunct to conventional therapy of coronary artery disease, including late phase after myocardial infarction or ischemic cardiomyopathy.

Hyperphosphataemia in renal failure: causes, consequences and current management.

Hyperphosphataemia is prevalent among chronic renal failure and dialysis patients. It is known to stimulate parathyroid hormone and suppress vitamin D3 production, thereby inducing hyperparathyroid bone disease. In addition, it may independently contribute to cardiac causes of death through increased myocardial calcification and enhanced vascular calcification. Hyperphosphataemia is also associated with cardiac microcirculatory abnormalities. Therefore, phosphate control is of prime importance. It is important to control phosphate levels early in the course of chronic renal failure in order to avoid and treat secondary hyperparathyroidism, and cardiovascular and soft tissue calcifications. Dietetic restrictions are often difficult to follow long term. Because of its large sphere of hydration and the complex kinetics of phosphate elimination, phosphate is not easily removed by dialysis. Long, slow dialysis may be effective, but this needs logistics and acceptance by patients. Thus, oral phosphate binders are generally required to control serum levels. None of the existing phosphate binding agents is truly satisfactory. Aluminium-containing agents are highly efficient but many clinicians have abandoned their use because of the potential toxicity. Despite of the wide use of calcium-containing agents, there was a link with hypercalcaemia and soft tissue calcifications. Novel phosphate binders in the form of polyallylamine hydrochloride, polyuronic acid derivatives and lanthanum carbonate appear promising. In this review, we discuss causes of hyperphosphataemia, pathological consequences and modalities of treatment.

Effect of Traumacel P on the growth of human dermal fibroblasts in vitro.

OBJECTIVE: It has been postulated that Traumacel P, a haemostatic powder, might assist the wound-healing process. This in vitro study investigated the effect of the powder on human dermal fibroblasts. METHOD: Experiments using fibroblasts from a 50-year-old donor were carried out using suspensions of Traumacel P ranging from 0.1 to 10.0 mg/ml. Dulbecco's Modified Eagle Medium with 10% or 0.4% foetal calf serum (FCS) and 5.5 mM or 25 mM glucose was used. The powder was either placed in contact with the cells or separated from them by a porous membrane. The response of the fibroblasts was assessed using the MTT assay. RESULTS: Concentrations of 0.5 mg/ml and 1.0 mg/ml stimulated the metabolic activity of the fibroblasts in both high and low glucose medium with 10% FCS, regardless of whether the powder was in contact with or separated from the cells. The greatest stimulation, to 174% of the controls, was produced by 0.5 mg/ml Traumacel P in low glucose/10% FCS with the powder in contact with the cells (p < 0.0001). Traumacel concentrations ranging from 0.5 to 5.0 mg/ml in 0.4% FCS significantly stimulated the metabolic activity of fibroblasts in low glucose medium, but not in the high glucose medium. CONCLUSION: These studies indicate that direct stimulation of fibroblast proliferation may be one of the ways in which Traumacel P aids the healing of ulcers observed in the clinic.

Can traumacel be used in the treatment of chronic wounds?

Traumacel (calcium salt of oxidised cellulose) has previously only been used in the treatment of acute wounds. To assess its safety and effectiveness in the management of chronic wounds a 12-week pilot study was undertaken which involved 11 patients with 15 non-healing leg ulcers and assessed ulcer size, ulcer pain and degree of exudate. Five ulcers healed within the study period. Significant pain relief was experienced by three patients. Use of the dressing did not appear to be related to a reduction in exudate. No patients experienced sensitisation to the product or had to be withdrawn because of adverse effects. Traumacel was found to be safe in the management of chronic ulceration, and appeared to promote healing in some recalcitrant ulcers.

Influence of Tolpa Peat Preparation on gastroprotection and on gastric and duodenal ulcers.

It was found, that Tolpa Peat Preparation administered prophylactically to rats decreased significantly the size of gastric mucosa damage induced with ethanol. TPP administered to rats with experimental gastric and duodenal ulcers significantly accelerated the healing process.

A randomised, double-blind study on the efficacy of Tolpa Torf Preparation (TTP) in the treatment of recurrent respiratory tract infections.

TTP is a new immunomodulating drug of natural origin, registered in Poland for human use. In the randomised, double blind study to assess the therapeutic efficacy of the Tolpa Torf Preparation (TTP) in the recurrent respiratory tract infectious participated 39 young patients (age 16-22). TTP was administered orally, 5 mg daily for three weeks. During 3 months follow-up period favourable results of treatment were obtained in 14 of 20 TTP treated patients and in 8 of 19 of the placebo patients. The therapeutic effects were seen even after the 6 months follow-up period. The phagocytic activity of granulocytes was significantly stimulated in the TTP--treated patients but not in the placebo-treated patients. The results suggest that TTP is effective drug in the treatment of recurrent respiratory tract infections with undefined infectious etiology. In the period of the treatment as well as during 6 months observation no side effects were noticed.

A comparison of efficacy of Tolpa Torf Preparation (TTP) in the treatment of cervicitis with or without surgery.

Tolpa Torf Preparation (TTP) is an immunomodulating drug produced by Torf Corporation, Wroclaw and registered for human use in Poland. TTP enhances the process of tissue regeneration. Authors evaluate TTP effectiveness in the treatment of inflammatory states of the cervix, especially cervical erosions and the influence of this preparation of the macroscopic, cytological and bacteriological state of the cervix. TTP was used in 31 patients with the diagnosis of cervical erosion. All patients treated as yet were classified into 3 groups, depending on the treatment of cervical erosion used previously. TTP was administered orally in the dose of 5 mg (in 10 ml of water) daily during 10 days and locally in the form of tampons soaked with 1% TTP solution in the volume of 5 ml also during 10 days. TTP administered this way has beneficial therapeutic effects on the healing of cervical erosion accelerating the process of epithelialization and bringing normalization of the cytological picture. Especially beneficial in the treatment of cervical erosion is combined use of TTP and electrocoagulation or curettage--the healing time can be shortened by half.

High-effective aluminium free phosphate binder. In vitro and in vivo studies.

For the purpose of intestinal phosphate binding we have developed aluminium free natural polymers consisting of heteropolyuronic acid charged with different cations. The in vitro experiments showed an efficacy two to three times greater than Aludrox. During up to six months of clinical application no serious side effects have been detected, constipation ceased and serum phosphate was maintained in an acceptable range.