Iguratimod ameliorates antibody-mediated rejection after renal transplant by modulating the Th17/Treg paradigm.

BACKGROUND: Iguratimod (IGU) is widely used in clinical practice due to its stable anti-inflammatory effects. Our previous studies have confirmed that the proportion of Th17/Treg balance in patients taking IGU altered significantly. This study aims to explore the role of IGU in antibody-mediated rejection (ABMR) and its potential mechanisms. METHODS: We conducted bioinformatics analysis of sequencing data from the GEO database to analyze the abundance of immune cell infiltration in transplanted kidney tissues. In vivo, IGU was intervened in a mice secondary skin transplantation model and a mice kidney transplantation ABMR model, and histological morphology of the grafts were examined by pathological staining, while relevant indicators were determined through qRT-PCR, immunohistochemistry, and enzyme-linked immunosorbent assay, observed T cell differentiation by flow cytometry, and preliminarily assessed the immunosuppressive effect of IGU. In vitro, we established Th17 and Treg cell induction and stimulation differentiation culture systems and added IGU for intervention to explore its effects on their differentiation. RESULTS: Through bioinformatics analysis, we found that Th17 and Treg may play important roles in the occurrence and development of ABMR. In vivo, we found that IGU could effectively reduce the damage caused by ABMR to the grafts, alleviate the infiltration of inflammatory cells in the graft tissues, and reduce the deposition of C4d in the grafts. Moreover, it is also found that IGU regulated the differentiation of Th17 and Treg cells in the spleen and peripheral blood and reduced the expression of IL-17A in the grafts and serum. In addition, same changes were observed in the induction and differentiation culture system of Th17 and Treg cells in vitro after the addition of IGU. CONCLUSION: IGU can inhibit the progression of ABMR by regulating the differentiation of Th17 and Treg cells, providing novel insights for optimizing clinical immunosuppressive treatment regimens.

A Robotic System With Embedded Open Microfluidic Chip for Automatic Embryo Vitrification.

Embryo vitrification is a fundamental technology utilized in assisted reproduction and fertility preservation. Vitrification involves sequential loading and unloading of cryoprotectants (CPAs) with strict time control, and transferring the embryo in a minimum CPA droplet to the vitrification straw. However, manual operation still cannot effectively avoid embryo loss, and the existing automatic vitrification systems have insufficient system reliability, and operate differently from clinical vitrification protocol. Through collaboration with in vitro fertilization (IVF) clinics, we are in the process realizing a robotic system that can automatically conduct the embryo vitrification process, including the pretreatment with CPAs, transfer of embryo to the vitrification straw, and cryopreservation with liquid nitrogen ( LN2). An open microfluidic chip (OMC) was designed to accommodate the embryo during the automatic CPAs pretreatment process. The design of two chambers connected by a capillary gap facilitated solution exchange around the embryo, and simultaneously reduced the risk of embryo loss in the flow field. In accordance to the well-accepted procedure and medical devices in manual operation, we designed the entire vitrification protocol, as well as the robotic prototype. In a practical experiment using mouse embryos, our robotic system showed a 100 % success rate in transferring and vitrifying the embryos, achieved comparable embryo survival rates (90.9 % versus 94.4 %) and development rates (90.0 % versus 94.1 %), when compared with the manual group conducted by the senior embryologist. With this study, we aim to facilitate the standardization of clinical vitrification from manual operation to a more efficient and reliable automated process.

Treatment of coking wastewater by an advanced Fenton oxidation process using iron powder and hydrogen peroxide.

In this study the treatment of coking wastewater was investigated by an advanced Fenton oxidation process using iron powder and hydrogen peroxide. Particular attention was paid to the effect of initial pH, dosage of H(2)O(2) and to improvement in biodegradation. The results showed that higher COD and total phenol removal rates were achieved with a decrease in initial pH and an increase in H(2)O(2) dosage. At an initial pH of less than 6.5 and H(2)O(2) concentration of 0.3 M, COD removal reached 44-50% and approximately 95% of total phenol removal was achieved at a reaction time of 1 h. The oxygen uptake rate of the effluent measured at a reaction time of 1h increased by approximately 65% compared to that of the raw coking wastewater. This indicated that biodegradation of the coking wastewater was significantly improved. Several organic compounds, including bifuran, quinoline, resorcinol and benzofuranol were removed completely as determined by GC-MS analysis. The advanced Fenton oxidation process is an effective pretreatment method for the removal of organic pollutants from coking wastewater. This process increases biodegradation, and may be combined with a classical biological process to achieve effluent of high quality.

Changes in biomass activity and characteristics of activated sludge exposed to low ozone dose.

In this paper, the response mechanism of activated sludge exposed to low-dose ozone at less than 20mgO(3)g(-1) total suspended solids (TSS) was studied by analyzing the changes in sludge activity and the evolution of C, N, P and metals from sludge following ozonation. The intracellular ATP concentration was not affected at less than 5mgO(3)g(-1) TSS and thereafter decreased rapidly to around 60% when the ozone dose increased to 20mgO(3)g(-1) TSS. Similarly, the efficiency of sludge solubilization initially changed a little and then increased rapidly to around 30% at an ozone dose of 20mgO(3)g(-1) TSS. However, the activities of superoxide dismutase and protease decreased immediately upon exposure to ozone. These findings indicate that ozone firstly destroys the floc, leading to the disruption of the compact aggregates, which does not affect cells viability but induces a decrease in enzyme activities. Ozone then attacks the bacterial cells of the sludge, causing a decrease in cells viability. During ozonation, the content of carbon, nitrogen and phosphorus in the sludge matrix decreased, while the content of these elements in the micro-solids and supernatant gradually increased. Most of the released metals from the sludge matrix were found in the micro-solids.

Systematic analysis of biochemical performance and the microbial community of an activated sludge process using ozone-treated sludge for sludge reduction.

Two lab-scale bioreactors (reactors 1 and 2) were employed to examine the changes in biological performance and the microbial community of an activated sludge process fed with ozonated sludge for sludge reduction. During the 122 d operation, the microbial activities and community in the two reactors were evaluated. The results indicated that, when compared with the conventional reactor (reactor 1), the reactor that was fed with the ozonated sludge (reactor 2) showed good removal of COD, TN and cell debris, without formation of any excess sludge. In addition, the protease activity and intracellular ATP concentration of reactor 2 were increased when compared to reactor 1, indicating that reactor 2 had a better ability to digest proteins and cell debris. DGGE analysis revealed that the bacterial communities in the two reactors were different, and that the dissimilarity of the bacterial population was nearly 40%. Reactor 2 also contained more protozoa and metazoa, which could graze on the ozone-treated sludge debris directly.

Progress and perspectives of sludge ozonation as a powerful pretreatment method for minimization of excess sludge production.

The treatment and disposal of excess sludge represents a bottleneck in wastewater treatment plants (WWTP) worldwide, due to environmental, economic, social and legal factors. The ideal solution to the problem of sludge disposal is to combine sludge reduction with the removal of pollution at the source. This paper presents an overview of the potential of ozonation in sludge reduction. The full-scale application of ozonation in excess sludge reduction is presented. Improvements in the biodegradability of the ozonated sludge were confirmed. The introduction of ozonation into activated sludge did not significantly influence effluent quality but improved the settling properties of the sludge. An operation with a suitable sludge wasting ratio seems to be necessary to prevent accumulation of inorganic and inert particles for long-term operation. Sludge ozonation to reduce excess sludge production may be economical in WWTP which have high sludge disposal costs and operational problems such as sludge foaming and bulking. The recommended ozone dose ranges from 0.03 to 0.05 g O(3)/g TSS, which is appropriate to achieve a balance between sludge reduction efficiency and cost. An effort to design and optimize an economic sludge reduction process is necessary.

Analysis of the mechanism of sludge ozonation by a combination of biological and chemical approaches.

Using the practical sludge obtained from municipal sewage treatment plants, the mechanism of the sludge ozonation process was systematically investigated by a combination of biological and chemical approaches, including analysis of the changes in biological response by CFU and PCR-DGGE, bio-macromolecular activity and radical scavenging activity. The results indicated that after the sludge was exposed to ozone at less than 0.02 g O(3)/g TSS, the DGGE fingerprint remained constant and there was still some enzyme activity, indicating that the sludge solubilization was the main process. At greater than 0.02 g O(3)/g TSS, the bacteria began to be broken down and ozone was used to oxidize the bio-macromolecules such as proteins and DNA released from the sludge. Bacteria belonging to 'G-Bacteria' were able to conserve their DNA in the presence of less than 0.08 g O(3)/g TSS. At levels higher than 0.10 g O(3)/g TSS, the disintegration of the sludge matrix became slow and the microbes lost most of their activity, and ozone was used to transform the bio-macromolecules into small molecules. However, at levels higher than 0.14 g O(3)/g TSS, the ozone failed to oxidize the sludge efficiently, because several radical scavengers such as lactic acid and SO(4)(2-) were released from the microbial cells in the sludge.

Enhanced sludge solubilization by microbubble ozonation.

A microbubble ozonation process for enhancing sludge solubilization was proposed and its performance was evaluated in comparison to a conventional ozone bubble contactor. Microbubbles are defined as bubbles with diameters less than several tens of micrometers. Previous studies have demonstrated that microbubbles could accelerate the formation of hydroxyl radicals and hence improve the ozonation of dyestuff wastewater. The results of this study showed that microbubble ozonation was effective in increasing ozone utilization and improving sludge solubilization. For a contact time of 80 min, an ozone utilization efficiency of more than 99% was obtained using the microbubble system, while it gradually decreased from 94% to 72% for the bubble contactor. The rate of microbial inactivation was obviously faster in the microbubble system. At an ozone dose of 0.02g O(3)g(-1) TSS, about 80% of microorganisms were inactivated in the microbubble system, compared with about 50% inactivation for the bubble contactor. Compared to the bubble contactor, more than two times of COD and total nitrogen, and eight times of total phosphorus content were released from the sludge into the supernatant by using the microbubble system at the same ozone dosage. The application of microbubble technology in ozonation processes may provide an effective and low cost approach for sludge reduction.

Enhanced ozonation of simulated dyestuff wastewater by microbubbles.

The ozonation of synthetic wastewater containing azo dye, CI Reactive Black 5, was investigated using a microbubble generator and a conventional bubble contactor. The microbubble generator produced a milky and high intensity microbubble solution in which the bubbles had a mean diameter of less than 58 microm and a numerical density of more than 2.9 x 10(4) counts ml(-1) at a gas flow rate of less than 0.5 l min(-1). Compared with the bubble contactor, the total mass transfer coefficient was 1.8 times higher and the pseudo-first order rate constant was 3.2-3.6 times higher at the same initial dye concentration of 100 mg l(-1), 230 mg l(-1) and 530 mg l(-1) in the proposed microbubble system. The amount of total organic carbon removed per g of ozone consumed was about 1.3 times higher in the microbubble system than in the bubble contactor. The test using terephthalic acid as the chemical probe implied that more hydroxyl radicals were produced in the microbubble system, which contributed to the degradation of the dye molecules. The results suggested that in addition to the enhancement of mass transfer, microbubbles, which had higher inner pressure, could accelerate the formation of hydroxyl radicals and hence improve the oxidation of dye molecules.

The expanded application of most probable number to the quantitative evaluation of extremely low microbial count.

This paper is about the evaluation of the extremely low microbial counts from the field by expanding the most probable number (MPN) methodology when the data follow Poisson distribution in order to achieve more accurate estimation with limited number of data. Several data sets with extremely low microbial counts from pharmaceutical clean room and laboratory clean bench monitoring were found with good Poisson distribution fitness by chi2 test (P < 0.05), and the MPN calculations were conducted for these data by using the Halvorson and Ziegler equation. The MPN values are generally larger than the arithmetic average, indicating a higher sensitivity of the data assay. This approach is justified because Poisson distribution is the mathematical background of the MPN procedure, with the Halvorson and Ziegler equation as its foundation. It is considered that the MPN methodology has a potential application for quality control in the extremely low level microbial counting environment in the clean rooms levels ISO class 7 or above. Further studies on the precision of this method and development of a sampling plan based on careful mathematical analysis will help to refine the approach.