OBJECTIVE: This study aimed to compare 9 perfluoroalkyl sulfonic acids (PFSA) with carbon chain lengths (C4-C12) to inhibit human placental 3β-hydroxysteroid dehydrogenase 1 (3β-HSD1), aromatase, and rat 3β-HSD4 activities. METHODS: Human and rat placental 3β-HSDs activities were determined by converting pregnenolone to progesterone and progesterone secretion in JEG-3 cells was determined using HPLC/MS-MS, and human aromatase activity was determined by radioimmunoassay. RESULTS: PFSA inhibited human 3β-HSD1 structure-dependently in the order: perfluorooctanesulfonic acid (PFOS, half-maximum inhibitory concentration, IC ₅₀: 9.03 ± 4.83 μmol/L) > perfluorodecanesulfonic acid (PFDS, 42.52 ± 8.99 μmol/L) > perfluoroheptanesulfonic acid (PFHpS, 112.6 ± 29.39 μmol/L) > perfluorobutanesulfonic acid (PFBS) = perfluoropentanesulfonic acid (PFPS) = perfluorohexanesulfonic acid (PFHxS) = perfluorododecanesulfonic acid (PFDoS) (ineffective at 100 μmol/L). 6:2FTS (1H, 1H, 2H, 2H-perfluorooctanesulfonic acid) and 8:2FTS (1H, 1H, 2H, 2H-perfluorodecanesulfonic acid) did not inhibit human 3β-HSD1. PFOS and PFHpS are mixed inhibitors, whereas PFDS is a competitive inhibitor. Moreover, 1-10 μmol/L PFOS and PFDS significantly reduced progesterone biosynthesis in JEG-3 cells. Docking analysis revealed that PFSA binds to the steroid-binding site of human 3β-HSD1 in a carbon chain length-dependent manner. All 100 μmol/L PFSA solutions did not affect rat 3β-HSD4 and human placental aromatase activity. CONCLUSION Carbon chain length determines inhibitory potency of PFSA on human placental 3β-HSD1 in a V-shaped transition at PFOS (C8), with inhibitory potency of PFOS > PFDS > PFHpS > PFBS = PFPS = PFHxS = PFDoS = 6:2FTS = 8:2FTS.
Humans ; Pregnancy ; Female ; Rats ; Animals ; Placenta ; Progesterone/pharmacology* ; Aromatase/pharmacology* ; Cell Line, Tumor ; Fluorocarbons ; Alkanesulfonic Acids ; Structure-Activity Relationship ; Hydroxysteroid Dehydrogenases/pharmacology*

Objective: The relationship between serum uric acid (SUA) levels and glycemic indices, including plasma glucose (FPG), 2-hour postload glucose (2h-PG), and glycated hemoglobin (HbA1c), remains inconclusive. We aimed to explore the associations between glycemic indices and SUA levels in the general Chinese population. Methods: The current study was a cross-sectional analysis using the first follow-up survey data from The China Cardiometabolic Disease and Cancer Cohort Study. A total of 105,922 community-dwelling adults aged ≥ 40 years underwent the oral glucose tolerance test and uric acid assessment. The nonlinear relationships between glycemic indices and SUA levels were explored using generalized additive models. Results: A total of 30,941 men and 62,361 women were eligible for the current analysis. Generalized additive models verified the inverted U-shaped association between glycemic indices and SUA levels, but with different inflection points in men and women. The thresholds for FPG, 2h-PG, and HbA1c for men and women were 6.5/8.0 mmol/L, 11.0/14.0 mmol/L, and 6.1/6.5, respectively (SUA levels increased with increasing glycemic indices before the inflection points and then eventually decreased with further increases in the glycemic indices). Conclusion An inverted U-shaped association was observed between major glycemic indices and uric acid levels in both sexes, while the inflection points were reached earlier in men than in women.
Aged ; Asian Continental Ancestry Group ; Blood Glucose/analysis* ; China/epidemiology* ; Cohort Studies ; Diabetes Mellitus/blood* ; Female ; Glucose Tolerance Test ; Glycated Hemoglobin A/analysis* ; Glycemic Index ; Humans ; Male ; Middle Aged ; Uric Acid/blood*

This paper aims to systematically analyze the peptides and proteins from Asini Corii Colla(ACC) through shotgun proteomics. After high-pH reversed-phase fractionation, the proteins and peptides in the hydrolysate of ACC were further separated by nano LC-Q-Exactive-MS/MS under the following conditions: Thermo Scientific EASY column(100 μm×2 cm, 5 μm, C_(18)) as precolumn, Thermo Scientific EASY column(75 μm×100 mm, 3 μm, C_(18)) for solid phase extraction, gradient elution with 0.1% formic acid in water(mobile phase A) and 84% acetonitrile in water containing 0.1% formic acid(mobile phase B), and MS in positive ion mode. Based on Uniprot_Equus caballus, MS data, and literature, 2 291 peptides were identified from ACC by MaxQuant, with 255 Maillard reactions(AML, CML, CEL)-modified peptides identified for the first time. Through alignment, the peptides were found to belong to 678 equine proteins. In conclusion, the combination of nano LC-Q-Exactive-MS/MS and shotgun proteomics achieved rapid and accurate identification of the proteins and peptides in ACC, which provides the key information and new insights for further investigation of chemicals and effective substances in ACC.
Animals ; Chromatography, Liquid ; Horses ; Peptides ; Proteins ; Proteomics ; Tandem Mass Spectrometry

Objective:To investigate the correlation between serum ferritin (SF) level and liver damage in the acute stage of dengue fever.Methods:A retrospective study was conducted to analyze 171 cases diagnosed with dengue fever as dengue fever group and 130 healthy patients as control group in Hangzhou 3A grade hospital from July to December 2017. Clinical data, SF and liver function related indicators were collected from both groups: alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL) to analyze the correlation between liver damage and SF in patients with dengue fever.Results:ALT, AST, and SF levels were significantly higher in the dengue fever group than those in the healthy control group ( Z = 11.553, 15.054 and 15.163, P < 0.001). SF levels were higher in the dengue fever combined with liver damage group than those without the liver damage group ( z = 6.930, P < 0.001). However, there was no statistically significant differences in age, gender, peak body temperature, and history of liver disease ( P > 0.05). In addition, Spearman’s correlation analysis showed that SF was positively correlated with ALT, AST, and TBIL ( r = 0.464, 0.531 and 0.315, P < 0.001). Among dengue patients with different SF levels, there were significant difference in ALT, AST levels and incidence of liver damage ( H = 14.240 and 17.584, χ2 = 49.547, P < 0.001). Patients with higher SF levels had higher ALT, AST levels and incidence of liver damage. Binary logistic regression analysis showed that hyperferritinemia (SF&ge;500 ng/ml) was the risk factor for dengue fever combined with liver damage ( OR = 8.120, P < 0.001). Furthermore, ROC curve analysis showed that the AUC for SF to judge dengue fever combined liver damage was 0.846 (95% CI: 0.785-0.908), and the sensitivity and specificity when the SF cut-off value was 1 506 ng/ml were 74.8% and 83.3%. Conclusion:There is a certain correlation between the SF level and the degree of liver damage in acute stage of dengue fever patients, and hyperferritinemia is a risk factor for dengue fever combined with liver damage.

OBJECTIVE: Severe hip osteoarthritis, caused by bone or joint maldevelopment, biomechanical transformation and previous surgical intervention, is inclusively existed in spondyloepiphyseal dysplasia (SED). To investigate and discuss the short-term efficacy and possible effects of total hip arthroplasty in the treatment of Tönnis grade 3 hip osteoarthritis in patients with SED. METHODS: From January 2017 to June 2019, 374 patients with hip osteoarthritis were involved for total hip arthroplasty conducted by senior professional surgeons, of whom 9 patients (6 males and 3 females) with 12 hip osteoarthritis secondary to the SED met the inclusive and exclusive criteria and received the above-mentioned hip operation. The short-term outcomes were observed. RESULTS: All the patients were implanted with Johnson & Johnson ceramic on ceramic cementless hip prostheses within the arthroplasty. They were followed up for an average period of 20 months. Except for one muscular calf vein thrombosis case, no complications, such as aseptic loosening, joint dislocation, fracture, neurovascular injury, deep vein thrombosis and infection were observed in all the 9 patients. Before the surgery, the average Harris hip score was 35.55, while the average of the Western Ontario and McMaster Universities osteoarthritis index (WOMAC) was 56.56. The level of quality of life indicated by SF-12 score was 41.56 on average. The mean pre-operation visual analogue scale (VAS) was 7.44. At the last follow-up, the average Harris hip score increased to 89.56, whereas the average WOMAC declined to 41.11. Compared with the baseline point, the average SF-12 score went up to 56.33. Dramatic drop of the mean VAS value to 2.67 was also observed at the last follow-up. In addition, post-operative increase of several pelvic-related parameters including pelvic incidence, pelvic tilt and sacral slope could be observed in the SED patients. The average measured pelvic incidence, pelvic tilt and sacral slope were 68.95°±4.60°, 52.75°±1.06° and 17.45°±1.77° before operation, respectively; whilst the mean value of these specific parameters increased to 76.98°±5.12°, 60.51°±4.35° and 18.10°±2.02°, respectively. The even leg lengths of the lower extremities were obtained after total hip arthroplasty. CONCLUSION Total hip arthroplasty is satisfactory in the short-term pain relieve and function recovery for the management of Tönnis grade 3 hip osteoarthritis secondary to the SED.
Arthroplasty, Replacement, Hip ; Female ; Follow-Up Studies ; Hip Prosthesis ; Humans ; Male ; Osteoarthritis, Hip/surgery* ; Osteochondrodysplasias ; Quality of Life ; Retrospective Studies ; Treatment Outcome

The H9N2 avian influenza virus ( AIV) has become increasingly concerning due to its role in severe economic losses in the poultry industry.Transmission of AIV to mammals, including pigs and humans, has accelerated to devise preventive strategies.To investigate the potential to be used as a vaccine vector for Eimeria mitis expressing antigens from H9N2 AIV, we here successfully developed stable transgenic E.mitis expressing HA1 protein from H9N2 AIV.Using the double-cassette expressing vector strategy with one cassette expressing yellow fluorescent protein ( YFP ) fused to muted dihydrofolate reductasethymidylate synthase derived from Toxoplasma gondii (TgDHFR—TSm2m3), the other expressing HA1 of the H9N2 virus, one transgenic E.mitis population ( Emi. HA1 ) was constructed.Sporozoites of E.mitis transfected with yellow fluorescent protein ( YFP) expression plasmid were inoculated into chickens via the cloacal route. The recovered fluorescent oocysts were sorted by fluorescence activated cell sorting ( FACS) and then successively passaged in chickens.The resulting population was analyzed by genome walking, western blot and indirect fluorescent assay ( IFA) . Genome walking confirmed the random integration of plasmid DNA into the genome; while western blot analysis demonstrated the expression of foreign protein-HA1.IFA result indicated the expressed by E.mitis mainly distributed the surface of cell membrane and the head of the sporozoites.We found that the reproduction of Emi.HA1 was similar with that of the parental strain.The expression of foreign antigens in the transgenic parasites will facilitate the development of transgenic E.mitis as a vaccine vector.

Weaning rabbits under the age of 3 months are most susceptible to coccidiosis, which is caused by the infection of 11 Eimeria species.However, no data is available about the susceptibility of rabbits under the age of 3 months.In this study, groups of rabbits at different ages (30 d, 50 d and 70 d) were infected with same dose of sporulated oocysts of Eimeria media or E.irresidua.Oocysts shed in feces during day 1 and day 5 after the infection were counted daily.The total number of oocysts production of 70 d old rabbits infected with E.media only was 3.7 ×107, obviously less than 30 d old rabbits (1.03 ×108) and 50 d rabbits (6.5 ×107).The same phenomenon was seen in rabbits infected with E.irresidua. At the same time, the clinical symptoms of the younger rabbits were obvious, reflecting depression, constipation and diarrhea.These results indicated that weaning rabbits at the age of 30 days are the most susceptible and thus provides clues to the control of coccidiosis in young rabbits.

Eimeria intestinalis is one relatively high pathogenic specie of rabbit coccidia.Fully understanding of its biological characteristics makes great contribution to the control of rabbit coccidiosis.In this study, the pathogenicity and immunogenicity of E.intestinalis ZJK strain was studied on weaning rabbits by investigating clinical syndrome, body weight loss and oocyst output.Four groups of rabbits were inoculated with sporulated oocysts in the dose of 0, 1 ×104 , 1 ×105 and 1 ×106, respectively.Then these rabbits were challenged with 1 ×106 oocysts 21 days post inoculation.The results show that the infection with 1 ×104 oocysts was sufficient to cause clinical syndrome and body weight loss, compared with the control group.And the groups given more oocysts show worse symptom.After challenge, no difference of the body weight gain was seen between non-infected control group and immunized groups; meanwhile, the oocyst output of the immunized rabbits was much lower than that of the non-immunized but challenged control group.These results suggest that E. intestinalis ZJK strain is of mediate pathogenicity and excellent immunogenicity. Therefore, a precocious line of E. intestinalis based on this strain can be a suitable candidate for the development of a live attenuated vaccine for rabbit coccidiosis.

Eimeria spp.of rabbits which cause great economic losses in rabbit industry belong to Phylum Apicomplexa. Since rabbit coccidia contain 11species, which have different pathogenicity between each other, it is necessary to identify the exact species in the feild samples to control rabbit coccidiosis.In order to improve the sensitivity and expand the application of PCR detection, we introduced the Whole Genome Amplification technique to preamplify the DNA template extracted from oocysts, then WGA product were used to amplify the ITS-1 sequence to identify species.The results showed that the optimized method guaranteed higher sensitivity which can identify the species based on single oocyst.Detection of 10 filed samples showed excellent specificity and consistency with morphological identification.By this method, the species that hard to discriminate under microscope can be accurately identified.Application of this method which provides higher sensitivity and specificity could make great effort to the rabbit coccidiosis investigation in practice.

Coccidiosis causes great economic losses to the rabbit industry.Currently, the most effective approach to control rabbit coccidiosis still relies on coccidiostats.This review summarized advances in related research at home and abroad on coccidiostats, such as sulfonamide, diclazuril, salinomycin, monensin.In addition, issues such as drug residue, as well as the appearance of drug-resistant strains were discussed, aiming to provide help for future control of rabbit coccidiosis.