Limited Clinical Efficacy with Potential Adverse Events in a Pilot Study of Autologous Adoptive Cell Therapy in Canine Oral Malignant Melanoma.

Adoptive cell therapy (ACT) has been studied in several human and canine cancers with some promising clinical outcomes but not in canine oral malignant melanoma (OMM). Our manuscript aimed to explore one kind of ACT, the ex vivo-expanded autologous immune cell infusion in canine OMM, as this tumor remains a treatment dilemma. The study recruited dogs with histopathological diagnoses of oral malignant melanoma, generated their peripheral blood mononuclear cells, expanded them into predominantly non-B non-T cells via stimulations of IL-15, IL-2, and IL-21, and then re-infused the cells into tumor-bearing dogs. Ten dogs were enrolled; three dogs did not report any adverse events; three had a mildly altered appetite; one had a mildly increased liver index, while the other three developed suspected anaphylaxis at different levels. The median progression-free interval was 49 days. Dogs with progressive disease during treatment had a shorter survival. This pilot study indicates limited efficacy with potential adverse events of this ACT. Most recruited patients were in a later stage and had macroscopic disease, which might affect the treatment efficacy. Further exploration of this cell therapy in an adjuvant setting, with adequate protocol modification and standardization, could still be considered.

Increased plasma DR-70 (fibrinogen-fibrin degradation products) concentrations as a diagnostic biomarker in dogs with neoplasms.

BACKGROUND: Tumor biomarkers have used widely in clinical oncology in human medicine. Only a few studies have evaluated the clinical utility of tumor biomarkers for veterinary medicine. A test for fibrinogen and fibrin degradation products (DR-70) has been proposed as an ideal biomarker for tumors in humans. The clinical value of DR-70 for veterinary medicine however has yet to be determined. OBJECTIVES: Investigate the diagnostic value of DR-70 concentrations by comparing them between healthy dogs and dogs with tumors. ANIMALS: Two hundred sixty-three dogs with different types of tumors were included. Sixty healthy dogs also were recruited for comparison. METHODS: The DR-70 concentrations were measured in all recruited individuals by ELISA. Clinical conditions were categorized based on histopathology, cytology, ultrasound examination, radiology, clinical findings, and a combination of these tests. RESULTS: The median concentration of DR-70 was 2.130 ± 0.868 µg/mL in dogs with tumors, which was significantly higher than in healthy dogs (1.202 ± 0.610 µg/mL; P < .0001). With a cut-off of 1.514 µg/mL, the sensitivity and specificity of DR-70 were 84.03% and 78.33%, respectively. The area under curve was 0.883. The DR-70 concentration can be an effective tumor biomarker in veterinary medicine. CONCLUSIONS AND CLINICAL IMPORTANCE: Increased DR-70 concentrations were not affected by tumor type, sex, age, or body weight. However, in dogs with metastatic mast cell tumors and oral malignant melanoma, DR-70 concentrations were significantly increased. Additional studies, including more dogs with nonneoplastic diseases, are needed to further evaluate the usefulness of DR-70 as a tumor biomarker.

Esterases From Bifidobacteria Exhibit the Conversion of Albiflorin in Gut Microbiota.

Bifidobacteria is an important microbe that inhabits the human gut. It is capable of metabolizing complex compounds in the human diet. Albiflorin, an antidepressant natural product from Radix Paeoniae Alba in China, is difficult to absorb after oral administration, and its metabolism has been proven to be closely related to the gut microbiota. In this study, we demonstrated in vitro that several Bifidobacteria species were able to convert albiflorin to benzoic acid, and four esterases (B2, B3, B4, and BL) from Bifidobacterium breve and Bifidobacterium longum were found through genome mining and modeled by SWISS-MODEL. B2 and B3 presented the strongest albiflorin metabolism ability. The optimal conditions, including temperature, buffer, and pH, for the conversion of albiflorin by the four esterases were investigated. Furthermore, the effect of esterase on the metabolism of albiflorin in vivo was confirmed by transplanting bacteria containing esterase B2. This study demonstrated the vital role of esterases from Bifidobacteria in the metabolism of natural compounds containing ester bonds, which could contribute to the development of new enzymes, microbial evolution, and probiotic adjuvant compounds for treatment.

Urinary angiostatin: a novel biomarker of kidney disease associated with disease severity and progression.

BACKGROUND: This study aimed to evaluate the value of urinary angiostatin levels for assessing disease severity and progression of IgA nephropathy (IgAN). METHODS: Urinary angiostatin was identified as one of the distinct proteins in samples of patients with IgAN analyzed by Raybiotech protein array, and further confirmed by enzyme-linked immunosorbent assay (ELISA). RESULTS: Urinary angiostatin levels were significantly higher in IgAN patients than that in healthy controls (HC) subjects and lower than in disease controls (DC) patients. The concentrations of angiostatin in urine normalized to urinary creatinine (angiostatin/Cr) were positively associated with proteinuria level. With advancing chronic kidney disease (CKD) stage, urinary angiostatin/Cr levels were gradually increased. Urinary angiostatin/Cr levels in patients with Lee's grade IV-V were significantly higher than those in Lee's grade I-II and III. We further compared urinary angiostatin/Cr levels by using Oxford classification and found the expression in patients with mesangial proliferative score 1(M1) was significantly higher than that in M0 (P < 0.001). In addition, the levels of urinary angiostatin/Cr in patients with tubular atrophy/interstitial fibrosis score 1(T1) and T2 were significantly higher than those in T0 (P < 0.01, P < 0.001, respectively). After follow-up, renal survival was significantly worse in patients with higher levels of urinary angiostatin (P < 0.05). CONCLUSIONS: Urinary angiostatin may be a useful novel noninvasive biomarker to evaluate disease severity and progression of IgAN.

The characteristics and risk factors for cisplatin-induced acute kidney injury in the elderly.

OBJECTIVES: The most serious adverse reaction of cisplatin is acute kidney injury (AKI). Cisplatin-induced acute kidney injury (CIA) has no specific preventive measures. This study aims to explore the characteristics and risk factors for CIA in the elderly and to identify potential methods to reduce CIA. MATERIALS AND METHODS: Patients ≥18 years old, with primary tumors, who received initial cisplatin chemotherapy and whose serum creatinine (SCr) values were measured within 2 weeks pre- and postcisplatin treatment and who had complete medical records, were selected from a single center from January 1, 2013 to December 31, 2015. The exclusion criteria included radiotherapy or surgery, recurrent tumors, previous cisplatin treatment, lack of any SCr values before or after cisplatin therapy, and incomplete medical records. RESULTS: Out of a total of 527 patients, 349 were elderly. Angiotensin-converting enzyme inhibitor/angiotensin receptor blocker (ACEI/ARB) use (9.2%) was more prevalent in the elderly than in younger patients (2.8%, p = 0.007). The dosage of cisplatin treatment was lower in the elderly, but the incidence of CIA (9.46%) was higher in the elderly than in younger patients (3.37%). There were significant differences in the SCr levels, estimated glomerular filtration rate, ACEI/ARB use, and whether a single application of cisplatin was administered, between the elderly AKI group and the non-AKI group. Multivariable analysis showed that administration of a single application of cisplatin (OR 2.853, 95% CI: 1.229, 6.621, p = 0.015) and ACEI/ARB use (OR 3.398, 95% CI: 1.352, 8.545, p = 0.009) were predictive factors for developing CIA in the elderly. CONCLUSION: The incidence of CIA in the elderly was higher than in younger patients. ACEI/ ARB usage and administration of a single application of cisplatin were independent risk factors for CIA in the elderly.

Efficacy and safety of calcineurin inhibitor treatment for IgA nephropathy: a meta-analysis.

BACKGROUND: IgA nephropathy is the most common progressive glomerular disease to end stage renal failure worldwide. Calcineurin inhibitors (CNIs) is a selective immunosuppressant widely used in organ transplantation. The efficacy and safety of calcineurin inhibitors for the treatment of IgA nephropathy remain uncertain. METHODS: We performed a systematic literature search using the PubMed, Embase, Science Citation Index, Ovid evidence-based medicine, Chinese Biomedical Literature (CBM) and Chinese science and technology periodicals (CNKI, VIP, and Wan Fang) for randomized, controlled trials of CNIs therapy of IgA nephropathy. Complete remission rate (CR) was defined as proteinuria less than 0.5 or 0.3 g/d. Partial remission rate (PR) was defined as proteinuria reduced to at least half of the baseline measurement and an absolute value of >0.5 or 0.3 g/d. RESULTS: Seven relevant trials were conducted with 374 patients enrolled. CNIs plus medium/low-dose steroid had a higher CR (RR = 2.51 [95% CI,1.25 to 5.04], P = 0.02) compared to therapy with steroid alone or placebo, but were not significant on PR (RR = 0.87 [95% CI,0.32 to 2.38]; P = 0.78). Also, significant alterations were observed in proteinuria (weighted mean difference, -0.46 g/d,[95% CI:-0.55 to -0.24], P < 0.01) with no differences were found in serum creatinine (SCr) (weighted mean difference, 0.57,95% CI:-4.05 to 5.19; P = 0.78) and estimated glomerular filtration rate (eGFR) (weighted mean difference, 1.13,95% CI:-4.05 to 6.32; P = 0.34) level between the two groups. CNI therapy was associated with an increased risk for adverse events (RR = 2.21,95% CI:1.52 to 3.21, P < 0.01), such as gastrointestinal and neurological symptoms or hirsutism. CONCLUSIONS: CNIs might provide renal protection in patients with IgAN, but at an increased risk of adverse events. Reliably defining the efficacy and safety of CNIs in IgAN requires a high-quality trial with a large sample size.

[Characteristics and Mechanism of Biological Nitrogen and Phosphorus Removal Granular Sludge Under Carbon Source Stress].

In SBR reactor, the mature granular sludge fed with sodium acetate was gradually cultivated with different carbon sources (sodium acetate/glucose ratio was 1:0, 3:1, 1:1, 1:3 and 0:1, in terms of COD, respectively). During the five stages, the physical, biochemical properties, extracellular polymeric substances (EPS), phosphorus fractions and nitrogen and phosphorus removal efficiency of granular sludge were studied. 705 days' experimental results were showed as follows. At stage Ⅳ, the granular sludge had the smallest diameter of 0.5 mm; moreover the phosphorus release/uptake rate, denitrification rate and the total phosphorus (TP) content were the lowest. While at stages Ⅰ and Ⅱ, the phosphorus release/uptake and denitrification rates were the highest, meanwhile, the TP content reached up to 72.36 mg·g-1, and the EPS content was about 350 mg·g-1, as a result, the nitrogen and phosphorus removal efficiencies were both over 94%. Nevertheless at stage Ⅴ, the biochemical rates were slightly slower than values of stages Ⅰ and Ⅱ, simultaneously the TP, glycogen and EPS contents in sludge were maintained at 69.60 mg·g-1, 224.18 mg·g-1 and 200 mg·g-1, respectively, while high nitrogen and phosphorus removal efficiency was obtained. During all stages, Ca-P was the main phosphorus fraction, and inorganic phosphorus(IP) was closely related to phosphorus removal of granular sludge.

[Simultaneous determination of salidroside and tyrosol in Beagle dog plasma using UHPLC-MS/MS after pre-column dansyl chloride derivatization].

A pre-column derivatization method combined with UHPLC-MS/MS was developed for the simultaneous determination of salidroside and tyrosol in Beagle dog plasma. After protein precipitation by acetonitrile, the liquid supernatant was treated with dansyl chloride under dark conditions at 60 ℃ for 30 min, and then, the sample solution was extracted using methyl tertiary butyl ether. The multiple reaction monitoring in positive ion mode was used for MS detection of the tested analytes with the specific ion transitions of m/z 534.2→372.0 for salidroside derivative, m/z 372.0→171.0 for tyrosol derivative and m/z 506.0→171.0 for arbutin derivative. The chromatograph separation was achieved on an ACQUITY UPLC® BEH C18 column (100 mm × 2.1 mm, 1.7 µm) with a gradient mobile phase consisting of acetonitrile (0.1% formic acid)-water (10% acetonitrile, 0.1% formic acid) for 9 min. The assay showed a good linearity over the range of 0.02/0.1 − 20/10 µmol·L−1 with a lower limit of quantitation of 0.02 and 0.1 µmol·L−1 for salidroside and tyrosol in dog plasma, respectively. The intra- and inter-day precisions were all less than 8.68%, and the accuracy was within ±11.4%. The established method with a high sensitivity, good specificity and reliability was appropriate for simultaneous determination of salidroside and tyrosol in dog plasma and successfully applied to a pharmacokinetic study after intragastric administration of salidroside to Beagle dogs.

[The Significance of Analysis of Soil from the Surface of Hetao Plain, Inner Mongolia with Near Infrared Spectra].

The technology of near infrared spectrum, marked by its convenience and effectiveness, which has been applied extensively in lithological analysis, is suitable for component analysis of regional geology, due to its superiority in analyzing numerous samples in a relatively short time. Since different mineral molecule peaks will show different characters when being analyzed in the near infrared spectrum, we can acquire the information about the mineral composition, water content, mineral crystallinity and etc. In this passage, the soil from the surface of Hetao Plain has been analyzed, and in this case, we can obtain the information about the mineral composition, water content, mineral crystallinity and etc. Referring the features of local environment and climate, we could surmise the source of the minerals in soil and the environment where they formed. The result not only consummates the geological characterization of Hetao area, but also has great significance revealing mechanism of soil-formation in Hetao area. The result shows that the soil from surface of Hetao Plain is mainly composed by altered minerals, such as kaolinite, smectite, ledikite, muscovite etc, and is presumed to be from acidic rock mass of the Yinshan mountains. The passage also indicates that the Yinshan mountains have great impact on the formation of the soil on Hetao Plain in the aspect of climate, for example, though the climate of inner Mongolia is continental arid climate, enough rainfall and the substantial differences in mean annual precipitation ensure the relatively moist environment in this area, and make the sediments altered adequately, proposing new aspects about exploring the mechanism of the soil-formation in Hetao area.

Simultaneous determination of evodiamine and evodine in Beagle dog plasma using liquid chromatography tandem mass spectrometry.

A sensitive, rapid, and specific liquid chromatography/tandem mass spectrometry assay has been established and validated for the quantitation of evodiamine and evodine in Beagle dog plasma. Plasma samples of 0.2 ml were processed by liquid-liquid extraction with n-hexane/ethyl acetate (2:1, v/v). Chromatographic separations were done on a Symmetry C18 column (100 mm × 4.6 mm, ID, 5 µm) at 35°C with a linear gradient of methanol and 20 mM ammonium formate containing 0.2% formic acid. Evodiamine, evodine, and glibenclamide [internal standard (IS)] were ionized with an electrospray ionization source operated in positive ion mode. The MS/MS transitions were m/z 304.1 â†’ 161.1 for evodiamine, m/z 471.2 â†’ 425.1 for evodine, and m/z 494.1 â†’ 369.1 for IS. Calibration curves were linear over the concentration range of 0.1-100 ng/ml for evodiamine and 0.5-500 ng/ml for evodine. The mean extraction recoveries were 88.10 ± 3.21% for evodiamine and 81.24 ± 4.07% for evodine. The intra- and inter-day precisions were less than 11.10% and 12.81%, and the accuracy was within ± 11.76% for both analytes. Evodiamine and evodine were stable during storage and analytical periods. The validated method has been successfully applied to a pharmacokinetic study of evodiamine and evodine in beagle dogs after oral administration.

[The relationship among congenital Toxoplasma gondii infection, pregnancy outcome and T lymphocyte subsets in umbilical cord blood].

OBJECTIVE: To reveal the relationship among congenital Toxoplasma gondii (T. gondii) infection, T lymphocyte cell subsets in umbilical cord blood and pregnancy outcome. METHODS: 784 umbilical cord blood samples were collected and information of pregnancy outcomes was collected in a hospital of Hefei city, Anhui province during March 2009 to May 2010. T. gondii IgM antibodies in the sera were detected by ELISA. For all neonates infected with T. gondii and 10 healthy neonates, T lymphocyte cell subsets were detected by flow cytometry. RESULTS: According to the detection results of T. gondii IgM antibodies, 784 neonates were divided into infection group (21 neonates) and control group (763 neonates). The body weight and 1 min Apgar score of infection group were (3116.4 ± 352.6) g and (8.21 ± 1.26) points, respectively, which were statistically lower than control group ((3220.1 ± 242.3) g and (8.77 ± 1.61) points, respectively) (P < 0.01). The proportion of adverse pregnancy outcome of infection group was 19.0% (4/21), which was statistically greater than control group (4.8%, 37/763) (P < 0.01). The percentage of CD(3)(+) T lymphocyte cells in umbilical cord blood in infection group with and without adverse pregnancy outcomes were (64.51 ± 5.27)% and (64.32 ± 4.56)%, respectively, which were statistically lower than control group ((69.32 ± 4.32)%) (P < 0.01). The ratio value of CD(4)(+)/CD(8)(+) in infection group with, without adverse pregnancy outcomes and control group are 1.39 ± 0.24, 1.64 ± 0.28 and 2.34 ± 0.46, respectively, which showed statistical difference between any 2 groups (P < 0.01). CONCLUSION: T. gondii infection leads to adverse pregnancy outcomes and disorder of cellular immunity while T lymphocyte cell subsets are closely associated with adverse pregnancy outcome.

[Development of a 2-level system of medical education during the period of the Republic of China].

The education system gradually improved with the development of school system reforms during the 1912 - 1927 period of the Republic of China, but there was no specific plan for the higher medical education system. A 2-level system of medical education was put forward according to the non-unified education system and social realities. After discussion, this system was eventually established. The mode of coexistence of elite and the public education was formed and the lack of medical personnel at that time was in part alleviated.