Characteristics, differential diagnosis, individualized treatment, and prevention of hyperhomocysteinemia in newborns.

OBJECTIVES: This study aimed to investigate the incidence rate, clinical phenotype, gene variation spectrum, and prognosis of neonatal hyperhomocysteinemia (HHcy) and explore its diagnosis, individualised treatment, and prevention strategies. METHODS: We screened 84722 neonates for HHcy using liquid chromatography-tandem mass spectrometry (LC-MS/MS) combined with biochemical detection, urine gas chromatography-mass spectrometry (GC-MS), and next-generation sequencing (NGS) for gene analysis to comprehensively differentiate and diagnose diseases. RESULTS: 18 children (P1-P18) were diagnosed with methylmalonic acidemia (MMA) and HHcy, and fourteen known and one new variant of the MMACHC gene were found. Five children showed poor mental reactions, brain dysplasia, lethargy, hyperbilirubinemia, and jaundice, whereas the other 13 children had no evident abnormalities. These children were all cobalamin- and folic acid-reactive types, and they were mainly supplemented with cobalamin, L-carnitine, betaine, and folic acid. The mother of P12 had a prenatal diagnosis at the next pregnancy; the results showed that MMACHC gene was not pathogenic and she gave birth to a healthy baby. One child (P19) was diagnosed with methylenetetrahydrofolate reductase (MTHFR) deficiency, and one new mutation was detected in the MTHFR gene. Patient P19 showed congenital brain dysplasia, neonatal anaemia, and hyperbilirubinemia, and treatment consisted mainly of betaine and cobalamin supplementation. One child (P20) was confirmed to have methionine adenosyltransferase I (MAT I) deficiency but had no clinical manifestations. After treatment, all the children had a good prognosis. CONCLUSION: The incidence of neonatal HHcy in the Zibo area was 1/4236, and the common pathogenic variants were c.609G>A, c.80A>G, and c.482G>A in the MMACHC gene. Patients with HHcy can achieve a good prognosis if pathogenic factors and targeted treatment are identified. Gene analysis and prenatal diagnosis contribute to the early prevention of HHcy.

Amniotic fluid karyotype analysis and prenatal diagnosis strategy of 3117 pregnant women with amniocentesis indication.

Aim: To examine prenatal diagnosis strategies through fetal karyotype analysis for 3117 pregnant women with genetic amniocentesis indications. Materials & methods: According to the different indications for amniocentesis, the study was divided into 8 groups. The number of amniocentesis specimens, the number of abnormal karyotypes and the positive rate of each group were analyzed. Results: Compared with prenatal serum screening, noninvasive prenatal DNA testing is more accurate and can effectively improve screening efficiency. Multiple prenatal diagnosis indicators (37.349%) were more likely to be detected than single prenatal diagnosis indicators (11.091%). Conclusion: None of the screening methods can completely replace amniocentesis, and for pregnant women with genetic indications for amniocentesis, amniocentesis is strongly recommended.

Application of Human Umbilical Cord Mesenchymal Stem Cells in Rat Spinal Cord Injury Model.

The treatment of spinal cord injury (SCI) is a hot topic in clinic. In this study, female rats were selected and randomly divided into four groups (normal, sham, SCI, and mesenchymal stem cells [MSCs] groups). Hemostatic forceps were used to clamp the spinal cord for 1 min to establish the SCI animal model in rats. The levels of proinflammatory factors in the blood of each group were compared 4 h after operation. The motor function of hind limb was estimated by Basso, Beattie & Bresnahan Locomotor rating scale (BBB scale) at 3 months after surgery, the spinal cord tissue from the experimental area was obtained and stained histologically and immunohistochemically. Basso, Beattie & Bresnahan Locomotor rating scale results indicated that human umbilical cord (HUC) MSCs transplantation could improve the walking ability in rats with the SCI. Human umbilical cord mesenchymal stem cells substantially upregulated the secretion of anti-inflammatory factors and downregulated the secretion of proinflammatory factors, and promoted the repair of the SCI and inhibited the increase of glial cells induced by the SCI. Human umbilical cord mesenchymal stem cells transplantation can partially recovered the motor ability of rats with the SCI through promoting the regeneration of nerve cell and the expression of neural related genes, and inhibiting inflammatory reaction.

Combining Bone Collagen Material with hUC-MSCs for Applicationto Spina Bifida in a Rabbit Model.

Spina bifida is one of the neural tube defects, with a high incidence in human birth defects, which seriously affects the health and quality of life of patients. In the treatment of bone defects, the source of autologous bone is limited and will cause secondary damage to the patient. At the same time, since the bone tissue in animals needs to play a variety of biological functions, its complex structure cannot be replaced by a single material. The combination of mechanical materials and biological materials has become a common choice. Human umbilical cord mesenchymal stem cells (hUC-MSCs) have the advantages of easy access, rapid proliferation, low immunogenicity, and no ethical issues. It is often used in the clinical research of tissue regeneration and repair. Therefore, in this study, we established a spina bifida model using Japanese white rabbits. This model was used to screen the best regenerative repair products for congenital spina bifida, and to evaluate the safety of regenerative repair products. The results showed that the combination of hUC-MSCs with collagen material had better regeneration effect than collagen material alone, and had no negative impact on the health of animals. This study provides a new idea for the clinical treatment of spina bifida, and also helps to speed up the research progress of regenerative repair products.

Combining Bone Collagen Matrix with hUC-MSCs for Application to Alveolar Process Cleft in a Rabbit Model.

BACKGROUND: Most materials used clinically for filling severe bone defects either cannot induce bone re-generation or exhibit low bone conversion, therefore, their therapeutic effects are limited. Human umbilical cord mesenchymal stem cells (hUC-MSCs) exhibit good osteoinduction. However, the mechanism by which combining a heterogeneous bone collagen matrix with hUC-MSCs to repair the bone defects of alveolar process clefts remains unclear. METHODS: A rabbit alveolar process cleft model was established by removing the bone tissue from the left maxillary bone. Forty-eight young Japanese white rabbits (JWRs) were divided into normal, control, material and MSCs groups. An equal volume of a bone collagen matrix alone or combined with hUC-MSCs was implanted in the defect. X-ray, micro-focus computerized tomography (micro-CT), blood analysis, histochemical staining and TUNEL were used to detect the newly formed bone in the defect area at 3 and 6 months after the surgery. RESULTS: The bone formation rate obtained from the skull tissue in MSCs group was significantly higher than that in control group at 3 months (P < 0.01) and 6 months (P < 0.05) after the surgery. The apoptosis rate in the MSCs group was significantly higher at 3 months after the surgery (P < 0.05) and lower at 6 months after the surgery (P < 0.01) than those in the normal group. CONCLUSIONS: Combining bone collagen matrix with hUC-MSCs promoted the new bone regeneration in the rabbit alveolar process cleft model through promoting osteoblasts formations and chondrocyte growth, and inducing type I collagen formation and BMP-2 generation.

Active bone material containing modified recombinant human bone morphogenetic protein-2 induces bone regeneration in rabbit spina bifida.

This study focuses on spina bifida, which is a high incidence among the current clinical manifestations of human birth defects. Because in the treatment of bone defects, the source of autologous bone is limited and it is easy to cause secondary injury to the patient. At the same time, since the bone tissue in animals needs to perform a variety of biological functions, its complex structure cannot be replaced by a single material. Therefore, in this study, we used Japanese white rabbits to establish an animal model similar to human congenital spina bifida. The established animal model is used to screen the best regenerative repair products for the treatment of congenital spondylolisthesis defects, and to evaluate the safety of regenerative repair products. The results show that bone morphogenetic protein (BMP)-2 combined with collagen material has a better regeneration effect than collagen material alone, and it did not negatively affect the health of animals. This study is not only suitable for the screening of large-scale biomaterials, accelerating the research progress of regenerative repair products, but also conducive to the research on the mechanism of regeneration and repair of various materials.

Newborn screening and genetic variation of medium chain acyl-CoA dehydrogenase deficiency in the Chinese population.

OBJECTIVES: Medium-chain acyl-CoA dehydrogenase deficiency (MCADD) is an autosomal recessive disorder of the fatty acid oxidative metabolism. This study aimed to investigate the epidemiological characteristics, the spectrum of variation, clinical phenotype, and prognosis of MCADD in Chinese newborns. METHODS: We retrospectively analysed newborn screening (NBS) data in the Zibo area from January 2016 to March 2022 and summarized 42 cases recently reported in Chinese neonates. High-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) and next-generation sequencing (NGS) were used to detect the concentrations of carnitine in the blood spots and for diagnosis. RESULTS: A total of 183,082 newborns were detected, and six patients were diagnosed with MCADD (1/3,0514). The primary octanoylcarnitine (C8) and the octanoylcarnitine/decanoylcarnitine ratio (C8/C10) were elevated in all patients. Gene analysis revealed four known and four novel variants of the ACADM gene. Five patients were asymptomatic and developed normally under dietary guidance. One child died of vaccination-induced MCADD, presenting with hypoglycemia and elevated acylcarnitines. CONCLUSIONS: The incidence of MCADD in Chinese newborns varies geographically from 1/222,903 to 1/30,514, and the most common pathogenic variant is c.449_452 del CTGA (p. T150Rfs∗4) in ACADM gene with a frequency of 27.7%. HPLC-MS/MS and genetic analysis are beneficial for early prevention and good prognosis of MCADD.

Genetically engineered FGF1-sericin hydrogel material treats intrauterine adhesion and restores fertility in rat.

Endometrial injury can cause intrauterine adhesions (IUA) and induce the formation of endometrial fibrosis, leading to infertility and miscarriage. At present, there is no effective treatment method for severe IUA and uterine basal injury with adhesion area larger than one-third of the uterus. In this study, we prepared FGF1 silk sericin hydrogel material (FGF1-SS hydrogel) to treat endometrial injury and prevent endometrial fibrosis. Compared with the silk sericin hydrogel material (WT-SS hydrogel), FGF1-SS hydrogel significantly promotes the cell migration and infiltration ability of endometrial stromal cells (ESCs). More importantly, FGF1-SS hydrogel can release FGF1 stably for a long time and inhibit the ESCs injury model forms fibrosis through the TGF-ß/Smad pathway. In the IUA rat model, FGF1-SS hydrogel treatment effectively restored the number of uterine glands and uterine wall thickness in rats, with a fertility rate of 65.1% ± 6.4%. The results show that FGF1-SS hydrogel is expected to be a candidate to prevent IUA.

Differences in Immunological Landscape between EGFR-Mutated and Wild-Type Lung Adenocarcinoma.

Recent clinical trials of lung adenocarcinoma with immune checkpoint inhibitors revealed that lung adenocarcinoma patients with EGFR mutations have a poor response to immunotherapy. However, the mechanisms have not been addressed. We performed immunohistochemistry analyses of resected lung adenocarcinoma tissues with and without EGFR mutations to investigate and compare the characteristics of the tumor microenvironment (TME). We retrospectively enrolled a total of 323 lung adenocarcinoma patients (164 had EGFR mutations), and their corresponding tissue samples were analyzed by the EGFR mutation test and immunohistochemistry. We selected the markers of the immune checkpoint molecule (PD1, PD-L1, and LAG-3) and immune cell (CD3, CD4, CD8, and Foxp3) as markers of the tumor microenvironment. Our results revealed that patients had a distinct tumor microenvironment between EGFR-mutant and wild-type lung adenocarcinomas; the expression of CD3, CD4, PD-L1, and Foxp3 in EGFR-mutant tumors was significantly higher than that in wild-type tumors, while the expression of LAG3 and PD-1 showed a positive correlation with EGFR-wild-type tumors. In survival analysis, EGFR-wild-type patients had longer disease-free survival (DFS) than EGFR-mutant patients (P = 0.0065). Our research demonstrates significant differences in tumor microenvironment composition between EGFR-mutant and wild-type patients. Our findings provide novel evidence that contributes to understanding the mechanism underlying the poor efficacy of immune checkpoint inhibitors.

Repair of alveolar cleft bone defects in rabbits by active bone particles containing modified rhBMP-2.

Objective: A model of alveolar cleft phenotype was established in rabbits to evaluate the effect of active bone particles containing modified rhecombinant human BMP-2 on the repair of the alveolar cleft. Methods: 2-month-old Japanese white rabbits were selected and randomly divided into four groups: normal, control, material and BMP groups. Blood biochemical analysis, skull tomography (microfocus computerized tomography), and histological and immunohistochemical staining analysis of paraffin sections were performed 3  and 6 months after operation. Results: Both types of collagen particles showed good biocompatibility and promoted bone regeneration. The effect of active bone particles on bone repair and regeneration was better than that of bone collagen particles. Conclusions: Active bone particles containing modified rhecombinant human BMP-2 can be used for incisors regeneration.

Active bone material containing modified recombinant human bone morphogenetic protein 2 induces bone regeneration in the alveolar process cleft in rabbits.

The clinical application of most materials used to fill severe bone defects is limited owing to the insufficient ability of such materials to induce bone regeneration over a long repair period. The purpose of this study was to establish a model for the alveolar process cleft in rabbits to evaluate the effect of active bone material in bone defect repair. The active bone material used in this study is a new bone repair material composed of a heterogeneous collagen membrane implanted with modified recombinant human bone morphogenetic protein 2. This proposed active bone material can specifically bind to collagen. Twenty-four young Japanese white rabbits (JWRs) were selected and randomly divided into four groups (normal, control, material, and bone morphogenetic protein groups). The alveolar process cleft model was established by removing an equal volume bone at the left maxillary position. Blood samples were collected from the JWRs 3 and 6 months after the surgery to evaluate the biocompatibility of the active bone materials. Subsequently, the skull model was established, and the appearance was observed. Imaging methods (including X-ray examination and micro-computerized tomography scanning), tissue staining, and immunohistochemistry were employed for the evaluation. The bone collagen material and active bone material exhibited high biocompatibility. In addition, the ability of the active bone material to induce bone repair and regeneration was higher than that of the bone collagen material. The active bone material exhibited satisfactory bone regeneration performance in rabbits, indicating its potential as an active material for repairing congenital alveolar process clefts in humans.

Repair of alveolar cleft bone defects by bone collagen particles combined with human umbilical cord mesenchymal stem cells in rabbit.

BACKGROUND: Alveolar cleft is a type of cleft lip and palate that seriously affects the physical and mental health of patients. In this study, a model of the alveolar cleft phenotype was established in rabbits to evaluate the effect of bone collagen particles combined with human umbilical cord mesenchymal stem cells (HUC-MSCs) on the repair of alveolar cleft bone defects. METHODS: A model of alveolar clefts in rabbits was established by removing the incisors on the left side of the upper jaw bone collagen particles combined with HUC-MSCs that were then implanted in the defect area. Blood biochemical analysis was performed 3 months after surgery. Skull tissues were harvested for gross observation, and micro-focus computerised tomography (micro-CT) analysis. Tissues were harvested for histological and immunohistochemical staining. The experiments were repeated 6 months after surgery. RESULTS: Bone collagen particles and HUC-MSCs showed good biocompatibility. Bone collagen particles combined with HUC-MSCs were markedly better at inducing bone repair and regeneration than bone collagen particles alone. CONCLUSIONS: Combining HUC-MSCs with bone collagen particles provides a simple, rapid and suitable method to fill a bone defect site and treat of alveolar cleft bone defects.

Comparison of three surgical models of bone tissue defects in cleft palate in rabbits.

OBJECTIVE: Cleft palate is one of the most common craniofacial birth defects in the maxillofacial region. There is an urgent need in tissue regeneration research to establish animal models that faithfully mimic human diseases. Here, we compared three surgical models of bone tissue defects in cleft palate in rabbits in order to screen for the biomaterials that induced optimal bone regeneration. DESIGN: Rabbits were used to establish the models of hard palate cleft, alveolar cleft, and alveolar process cleft. Eight weeks following surgery, bone tissue self-healing capacity was estimated by macroscopic appearance and calculating the area of defective bone tissue. The dimensions of the upper jaw in left and right sides were measured at zero and eight weeks. RESULTS: Bone defects in three types of cleft palate models were made at the positions of the hard palate, alveoli and alveolar process. After 8 weeks, when the hard palate was partially excised, it underwent self-healing. When the hard palate was completely excised, it underwent partial self-healing. However, in the models of alveolar cleft and alveolar process cleft, there was no significant self-healing in the bone tissues. The dimensions of the upper jaw in left and right sides were no significant differences in three types of cleft palate models. CONCLUSIONS: Bone defects in the alveolar and alveolar process clefts exhibit a diminished capability for self-healing. This study may provide valuable information for the screening of materials that induce bone regeneration.

[Effects of Wastewater Nitrogen Concentrations and NH4+/NO3- on Nitrogen Removal Ability and the Nitrogen Component of Myriophyllum aquaticum (Vell.) Verdc].

Solution culture experiments were conducted to investigate the effect of wastewater nitrogen levels and NH4+/NO3- on nitrogen removal ability and the nitrogen component of Myriophyllum aquaticum. Experiments with three nitrogen levels and NH4+/NO3- were set up as follows:20, 100, and 200 mg·L-1and NH4+/NO3- 1:0, 0.5:0.5, and 0:1. The results showed that the biomass of plants increased fastest during the first week. The plants treated with NH4+/NO3-=1:0 with nitrogen levels of 20 and 100 mg·L-1 and those treated with NH4+/NO3-=0.5:0.5 with a nitrogen concentration of 200 mg·L-1 exhibited higher biomass than the others. The removal rates of water total nitrogen, ammonium nitrogen, and nitrate nitrogen during the first week were the maximum for all treatments and increased with water nitrogen levels. There were no significant differences in the removal rate between ammonium nitrogen and nitrate nitrogen with a nitrogen level of 20 mg/L, while with nitrogen levels of 100 and 200 mg·L-1, the nitrate removal rates were higher than those for ammonium nitrogen. The Myriophyllum aquaticum nitrogen accumulation and its contribution rate to nitrogen removal from water and sediment were all increased with water nitrogen levels and increased fastest during the first week. The contribution rate of nitrogen accumulated by plants with NH4+/NO3-=0:1 was the highest with nitrogen levels of 20 mg·L-1, while plants with NH4+/NO3-=0.5:0.5 were the highest with nitrogen levels of 100 and 200 mg·L-1. The protein, amino, and nitrate nitrogen contents in Myriophyllum aquaticum plants were all increased by increasing water nitrogen levels with a ranking of protein content > amino nitrogen content > nitrate nitrogen content. The protein concentrations in plants with NH4+/NO3-=1:0 and NH4+/NO3-=0.5:0.5 were higher regardless of water nitrogen levels, while the amino nitrogen concentration in plants with NH4+/NO3-=1:0 and the nitrate nitrogen content in plants with NH4+/NO3-=0:1 were higher than the others. It was concluded that the nitrogen removal ability of Myriophyllum aquaticum was improved by raising water nitrogen levels under the tested condition, which indicates that Myriophyllum aquaticum could purify high nitrogen wastewater. Myriophyllum aquaticum is an ammonium-nitrophile, but had the strongest capacity for growing and removing wastewater nitrogen exhibited with higher than 100 mg·L-1 nitrogen levels only with equal NH4+ to NO3-. The nitrogen component concentrations of protein, amino, and nitrate in Myriophyllum aquaticum plant were all affected by the ratio of NH4+/NO3-.

Thiazolidinediones and risk of colorectal cancer in patients with diabetes mellitus: A meta-analysis.

Background/Aims: A growing body of evidence has suggested that thiazolidinediones (TZDs) potentially reduce the risk of colorectal cancer (CRC). This study aimed to evaluate the effect of TZDs on CRC risk in patients with diabetes mellitus (DM). Patients and Methods: A systematic search of electronic databases was performed for studies evaluating the exposure to TZDs and reporting CRC risk in diabetic patients. Pooled estimates with 95% confidence intervals (CIs) were estimated using fixed or random effects models. Results: A total of 10 observational studies reporting more than 18,972 CRC cases in 2,470,768 DM patients were included. Meta-analysis showed a 9% reduction in CRC risk associated with TZDs use in all studies [relative risk (RR) =0.91, 95% CI = 0.84-0.99, P = 0.03] and cohort studies (RR = 0.89, 95% CI = 0.80-0.99, P = 0.04), respectively. However, such effect was not shown in case-control studies. In subgroup analyses, lower CRC risk was found in Asian population (RR = 0.40, 95% CI = 0.29-0.53, P = 0.00), and a trend toward lower CRC risk was observed in US population (RR = 0.94, 95% CI = 0.88-1.01, P = 0.08). CRC risk was significantly modified with non-pioglitazone TZD use (RR = 0.88, 95% CI = 0.82-0.95, P = 0.00), but not with pioglitazone use (RR = 0.95, 95% CI = 0.89-1.01, P = 0.11). No significant difference was observed with cancer site (colon or rectum). There was considerable inherent heterogeneity across studies, partly explained by study location. Conclusions: This meta-analysis supports a protective association between TZDs use and CRC risk in patients with DM. Future well-designed prospective studies with larger cohorts would be needed to understand this association better.

Fibrinogen-like protein 2 prothrombinase may contribute to the progression of inflammatory bowel disease by mediating immune coagulation.

Inflammation and coagulation are interdependent processes that enable each process to activate and propagate the other in inflammatory bowel disease (IBD). Thus, we investigated the role of a novel immune coagulant, fibrinogen-like protein 2 prothrombinase (FGL2), in patients and mice with IBD. 83 IBD patients and 40 normal controls were enrolled, and trinitro-benzene-sulfonic acid (TNBS)-induced colitis mice were used. Expression of FGL2 in the intestine was detected by immunohistochemistry. Using serial sections, staining was performed to detect tumor necrosis factor α (TNF-α) expression, and to demonstrate co-localization of FGL2 with macrophages and fibrin. Correlations between FGL2 expression with some common laboratory parameters were examined. FGL2 was seen primarily in inflammatory infiltrating cells, mainly macrophages, and microvascular vessels and had a strong co-localization with fibrin deposition. IBD patients and mice had increased expression of FGL2 compared with controls. Furthermore, FGL2 expression was correlated with intestinal and plasmatic TNF-α expression, mean platelet volume (MPV), platelet count (PLT), platelet-crit (PCT), and fibrinogen. Our data indicate that FGL2 may mediate immune coagulation in IBD patients. It may be considered as a novel molecule that contributes to the onset and development of IBD.

Uptake, transport and distribution of molybdenum in two oilseed rape (Brassica napus L.) cultivars under different nitrate/ammonium ratios.

OBJECTIVES: To investigate the effects of different nitrate sources on the uptake, transport, and distribution of molybdenum (Mo) between two oilseed rape (Brassica napus L.) cultivars, L0917 and ZS11. METHODS: A hydroponic culture experiment was conducted with four nitrate/ammonium (NO3-:NH4+) ratios (14:1, 9:6, 7.5:7.5, and 1:14) at a constant nitrogen concentration of 15 mmol/L. We examined Mo concentrations in roots, shoots, xylem and phloem sap, and subcellular fractions of leaves to contrast Mo uptake, transport, and subcellular distribution between ZS11 and L0917. RESULTS: Both the cultivars showed maximum biomass and Mo accumulation at the 7.5:7.5 ratio of NO3-:NH4+ while those were decreased by the 14:1 and 1:14 treatments. However, the percentages of root Mo (14.8% and 15.0% for L0917 and ZS11, respectively) were low under the 7.5:7.5 treatment, suggesting that the equal NO3-:NH4+ ratio promoted Mo transportation from root to shoot. The xylem sap Mo concentration and phloem sap Mo accumulation of L0917 were lower than those of ZS11 under the 1:14 treatment, which suggests that higher NO3-:NH4+ ratio was more beneficial for L0917. On the contrary, a lower NO3-:NH4+ ratio was more beneficial for ZS11 to transport and remobilize Mo. Furthermore, the Mo concentrations of both the cultivars' leaf organelles were increased but the Mo accumulations of the cell wall and soluble fraction were reduced significantly under the 14:1 treatment, meaning that more Mo was accumulated in organelles under the highest NO3-:NH4+ ratio. CONCLUSIONS: This investigation demonstrated that the capacities of Mo absorption, transportation and subcellular distribution play an important role in genotype-dependent differences in Mo accumulation under low or high NO3-:NH4+ ratio conditions.

[Effect of Wastewater Nitrogen Concentrations on Nitrogen Removal Ability of Myriophyllum aquaticum].

Myriophyllum aquaticum, which is an important plant for constructed wetlands, has powerful purification ability for wastewater, however, the relationship between nitrogen removal ability of Myriophyllum aquaticum and wastewater nitrogen concentrations is still unclear. In this study, pot culture experiment was conducted to investigate the effect of wastewater nitrogen levels on nitrogen removal ability of Myriophyllum aquaticum. 7 nitrogen levels were set up as following:2, 5, 10, 20, 100, 200, 400 mg·L-1. The results showed that when the wastewater nitrogen concentration was not higher than 20 mg·L-1, Myriophyllum aquaticum with 20 mg·L-1 of nitrogen concentration grew best in the first 3 weeks; the removal rates of total and ammonia nitrogen were nearly 100% after one week, while the nitrate nitrogen concentrations were very low and varied little; the nitrogen contents of Myriophyllum aquaticum had no significant change, the upper part nitrogen content was higher than the underneath, Myriophyllum aquaticum could also remove nitrogen from the sediment. When wastewater nitrogen concentrations were 100-400 mg·L-1, Myriophyllum aquaticum with 200 mg·L-1 of nitrogen concentration grew best from 4th to 5th week; the removal rates of total nitrogen were 76.5%, 71.5% and 48.1% in the three treatments, and the removal rates of ammonia nitrogen were 99.6%, 99.3% and 60.2% respectively, while the removal rates of nitrate nitrogen were all about 50% and there was no significant difference among treatments; the nitrogen contents of Myriophyllum aquaticum increased with nitrogen levels, but the difference between upper part and underneath was not remarkable, showing uniform distribution; nitrogen accumulations by Myriophyllum aquaticum and sediment accounted for 27.9%-48.4% and 12.2%-24.4% of total nitrogen loss in wastewater. Therefore, the nitrogen removal ability of Myriophyllum aquaticum should be inhibited by higher wastewater nitrogen level, the ammonia nitrogen removal rate was significantly higher than nitrate, the mechanism of Myriophyllum aquaticum nitrogen accumulation and distribution should also be affected by wastewater nitrogen level, and further research is needed.

Synovial Fluid Macrophage Migration Inhibitory Factor Levels Correlate with Severity of Self-Reported Pain in Knee Osteoarthritis Patients.

BACKGROUND Inflammation is considered as one of the main pathogeneses in OA-induced pain. Macrophage migration inhibitory factor (MIF) is a well known pro-inflammatory cytokine. We aimed to determine whether MIF levels in serum and synovial fluid (SF) are associated with severity of OA-induced pain. MATERIAL AND METHODS We recruited 226 patients with knee OA and 106 controls. Self-reported pain severity of OA patients was evaluated using the Western Ontario McMaster University Osteoarthritis (WOMAC) pain scores. MIF levels were detected using enzyme-linked immunosorbent assay (ELISA). RESULTS OA patients had similar serum MIF levels compared to controls (11.93 [5.68-18.10] vs. 10.06 [6.60-14.61] ng/ml, P>0.05). In OA patients, MIF levels in SF were dramatically lower compared to paired serum samples (3.39 [1.87-5.89] vs. 11.93 [5.68-18.10] ng/ml, P<0.01). MIF levels in SF were significantly correlated with WOMAC pain scores (r=0.237, P<0.001), but MIF levels in serum had no significant correlation with WOMAC pain scores (r=0.009, P=0.898). CONCLUSIONS MIF levels in SF, but not in serum, were independently associated with the severity of self-reported pain in OA patients. The inhibition of MIF signaling pathways may be a novel therapeutic approach for ameliorating OA-induced pain.

A giant magnetoimpedance-based biosensor for sensitive detection of Escherichia coli O157:H7.

A giant magnetoimpedance (GMI)-based biosensor was developed for detection of Escherichia coli (E. coli) O157: H7. The GMI sensor involving the sensing elements of Cr/Cu/NiFe/Cu/NiFe was fabricated by Micro Electro-Mechanical system (MEMS) technology, including thick photoresist lithography and electroplating. A separate Au film substrate as immunoplatform was used to capture E. coli O157:H7. The monoclonal mouse anti-E. coli antibody was immobilized on Au film substrate surface with a self-assembled layer. The different concentration E. coli O157:H7 (100, 300 and 500 cfu/ml) were combined with Dynabeads-antibody conjugates (DAC) respectively. DAC were prepared by conjugating streptavidin-coupled Dynabeads with biotin-labeled polyclonal mouse anti-E. coli antibodies. The classical sandwich assay was used for detection of E. coli O157:H7 targeted with Dynabeads by using antibody-antigen pair combination of biotin-streptavidin. The fundamental principle for detection of E. coli O157:H7 based on GMI sensor was that Dynabeads were employed as magnetic labels of E. coli O157:H7, and E. coli O157:H7 can be monitored by detecting the fringe field of Dynabeads using magnetic sensing elements. We observed that the GMI ratio were significantly improved due to the presence of E. coli O157:H7 combined with Dynabeads. The GMI ratio increased as the E. coli O157:H7 concentration increased. A lower detectable concentration of 100 cfu/ml was achieved in present work. The GMI-based biosensor provides a new method to rapid and sensitive detection E. coli O157:H7, which has a large potential for bio-application.