To optimize treatment for rheumatoid arthritis (RA), it is ideal to monitor the disease activity on a daily basis because RA activity fluctuates over time. Urine can be collected routinely at home by patients. Recently, we identified four urinary biomarker candidates—gelsolin (GSN), orosomucoid (ORM)1, ORM2 and soluble CD14 (sCD14)—in RA patients through transcriptomic and proteomic studies. Here, we investigated the clinical significance of the aforementioned urinary biomarker candidates in a prospective manner. For the first time, we found that urinary ORM1, ORM2 and sCD14 levels, but not GSN, were elevated in RA patients and had a positive correlation with the status of the disease activity. In particular, urine tests for ORM 1, ORM 2 and sCD14 efficiently represented the presence of high RA activity without the need for measuring blood markers. In a parallel study, a more rapid radiographic progression over 3 years was observed in patients with higher ORM2 levels. Combined measurements of urinary ORM2 and serum C-reactive protein synergistically increased the predictability of the radiographic progression of RA (odds ratio: 46.5). Collectively, our data provide evidence that blood-free, urinary biomarkers are promising surrogates for assessing disease activity and prognosis of RA. We anticipate that our urinary biomarkers will provide novel candidates for patient-driven measurements of RA activity at home and can shift the paradigm from blood to urine testing in the assessment of RA activity and prognosis in hospitals.
Arthritis, Rheumatoid* ; Biomarkers* ; C-Reactive Protein ; Humans ; Orosomucoid ; Prognosis* ; Prospective Studies

AIMTo discuss the relationship between the three-point interaction principle in the stereoselective separation of chromatography and applying this principle to survey the infrared spectrometry of racemate.

METHODSIn proving the applicability of the three-point interaction principal in IR spectrometry, a special case was found that phenylglycine did not obtain enantioselective separation on the chiral column but its IR spectrometry still obey this principle and explained such special case by experiment.

RESULTSAfter an equal quantity of solid crystal of d-phenylglycine and l-phenylglycine were mixed and ground for several minutes, they transformed to racemic compound. X-powder diffraction also confirmed this fact.

CONCLUSIONThe three-point principle was relatively reliable when it was used in the enantioselective chromatography separation and the IR spectrometric analysis. The reason of the fact that phenylglycine was not separated by chiral column can be explained by the fact that the acting force between the three polarity groups in the enantiomers is so strong that they can not form the instantaneous diastereoisomer with the chiral column, it was agreeable with the phenomenon that racemic mixture easily became racemic compound only by simply grinding the mixture in IR spectrometric experiment.

Chromatography, High Pressure Liquid ; Glycine ; analogs & derivatives ; analysis ; chemistry ; Orosomucoid ; chemistry ; Spectrophotometry, Infrared ; Stereoisomerism

To study the effect of alpha1-acid glycoprotein 1 (ORM1) polymorphism on the concentration of free nortriptyline in serum, genotyping analysis was employed in ORM1 by sequencing. Eighteen unrelated male adults were chosen and given a single dose of 25 mg nortriptyline orally, then the blood samples were taken at 0, 1, 2, 3, 4, 6, 8, 12, 24, 32, 48, 72, 96 and 168 hours after drug administration. Nortriptyline and 10-OH-nortriptyline in serum and ultrafiltrate were detected for the total and free concentration by using HPLC-MS/MS. Pharmacokinetic parameters were compared among different ORM1 genotypes. No significant differences were shown in the pharmacokinetic parameters of total nortriptyline and 10-OH-nortriptyline. The mean AUC(0-infinity) of free nortritpyline in ORM1 * F/ * F1 subjects was significantly higher than that in ORM1 * F1/ * S and ORM1 * S/ * S subjects [(119.1 +/- 74.4) ng x mL(-1) x h vs (51.4 +/- 23.2) ng x mL(-1) x h and (42.4 +/- 11.6) ng x mL(-1) x h]. The percentage of protein binding in subjects with ORM1 * F1/ * F1 genotype at 2, 3, 4, 6, 8 and 12 h after administration was slightly lower than in those with ORM1 * F1/ * S and ORM1 * S/ * S genotypes while the distinct difference was shown at 4 h (P < 0.05). Different ORM1 genotypes might affect the protein binding percentage and the concentration of serum free nortriptyline. The ability binding to the drug was higher in subjects with ORM1 * S/ * S genotype than in those with other two genotypes, so as to cause the lower concentration of free nortriptyline.
Adult ; Area Under Curve ; Genotype ; Humans ; Male ; Nortriptyline ; analogs & derivatives ; blood ; pharmacokinetics ; Orosomucoid ; genetics ; metabolism ; Polymorphism, Genetic ; Protein Binding

A new MS-titration method for the non-covalent binding of protein-ligand based on the research of berberine and alpha1-acid glycoprotein was established. The major presumption of new method is that the total concentration of protein-ligand complex is approximately the same as the total concentration of acting protein if a certain extent of affinity is existed between protein and ligand, in addition, the mole amount of acting ligand is more than that of acting protein. The non-covalent binding behaviours between berberine and alpha1-acid glycoprotein was studied by using electrospray ionization ion trap mass spectrometry (ESI-ITMS) , and the results were verified by fluorescence quenching method. The results showed that the binding behaviours between berberine and alpha1-acid glycoprotein, for example, stability constant, number of binding site, type of the main binding force, were almost the same by using the new MS-titration method and fluorescence quenching method. Comparing with the reported MS-titration method, the presented MS-titration method in this paper is more simple and applicable, does not demand much for the devices, and can lead to reliable results in same cases.
Berberine ; metabolism ; Ligands ; Orosomucoid ; metabolism ; Protein Binding ; Protein Interaction Mapping ; methods ; Proteins ; metabolism ; Spectrometry, Mass, Electrospray Ionization

BACKGROUND: Depletion of nutritional reserves and significant weight loss can lead to an increased risk of morbidity, reduced chemotherapy response, and shorter survival in patients with cancer. Numerous methodologies are used for the assessment of nutritional status. However, it remains unclear which of these tools is the most appropriate in the setting of cancer chemotherapy. The PINI (prognostic inflammatory nutritional index) is a simple scoring system that has been used to evaluate nutritional status and prognosis in critically ill patients. The aim of this study was designed to investigate the relationship between nutritional status after the palliative chemotherapy and prognosis in advanced metastatic cancer. METHODS: This study included 130 patients with ECOG PS (Eastern Cooperative Oncology Group performance status) 1~3 in advanced metastatic cancer following the palliative chemotherapy. ECOG PS, body mass index, alpha 1-acid glycoprotein, C-reactive protein, albumin, prealbumin, transferrin, protein, lactate dehydrogenase, PINI ratio, and survival time were evaluated. RESULTS: In 130 patients, the median age was 57 years. The ECOG PS was 1 68 patients, 2 43, 3 19. The mean value of PINI was PS 1 16.0+/-47.8, PS 2 55.6+/-106.9, PS 3 106.3+/-141.6. The mean survival was PS 1 144.0+/-71.8, PS 2 68.9+/-41.2, PS 3 32.0+/-6.9 days. In the PS 3 group, the mean values of alpha 1-acid glycoprotein, C-reactive protein, and lactate dehydrogenase were significantly higher than in the PS 1 group. In the PS 3 group, the mean values of albumin, prealbumin, and transferrin were significantly lower than in the PS 1 group. The survival time was correlated with the ECOG PS (r=-0.602, p<0.001,), PINI (r=-0.318, p<0.001,), alpha 1-acid glycoprotein (r=-0.265, p=0.002), C-reactive protein (r=-0.345, p<0.001), albumin (r=0.324, p<0.001), prealbumin (r=0.260, p=0.003) and transferrin (r=0.277, p=0.001). CONCLUSIONS: The PINI may be a useful scoring system for the assessment of nutritional status and prognosis in advanced metastatic cancer following palliative chemotherapy, but the ECOG PS is most strong correlation with the survival time.
Body Mass Index ; C-Reactive Protein ; Critical Illness ; Drug Therapy ; Humans ; L-Lactate Dehydrogenase ; Nutrition Assessment ; Nutritional Status* ; Orosomucoid ; Prealbumin ; Prognosis* ; Transferrin ; Weight Loss

AIMTo study the chromatographic behavior of cetirizine dihydrochloride on the proteinate- and amylose- based chiral stationary phases so as to optimizate the chromatographic condition of its enantiomers separation.

METHODSWhen using amylose-based, alpha1-acid glycoprotein and ovomucoid protein chiral stationary phase, the mobile phase was hexane-isopropyl alcohol-alcohol-trifluoroacetic acid (430:45:25:1), acetonitrile-10 mmol x L(-1) phosphate buffer solution (adjusted to pH 7.0 with sodium hydroxide) (4:96) and acetonitrile-20 mmol x L(-1) KH, PO4 solution (adjusted to pH 7.0 with triethylamine) (12.7:87.3), respectively. The temperature of proteinate column was 25 degrees C. The detective wavelength was 230 nm.

RESULTSThe two enantiomers could be separated on the two kinds of chiral stationary phases without derivatization and the resolution was above 2.0. The methods developed on the two kinds of chiral stationary phases are accurate, sensitive and specific.

CONCLUSIONBoth the proteinate- and amylose-based chiral stationary phases can be used to separate the enantiomers of cetirizine.

Amylose ; analogs & derivatives ; Cetirizine ; chemistry ; isolation & purification ; Chromatography, High Pressure Liquid ; Histamine H1 Antagonists, Non-Sedating ; chemistry ; isolation & purification ; Molecular Structure ; Orosomucoid ; Stereoisomerism

Rutin deca (H-) sulfate sodium (RDS) possesses very good activity as an inhibitor of the complement system of warm-blooded animals and HIV. An ion-pair coupled with solid-phase extraction technique (IP-SPE) was developed to extract RDS from rat plasma, urine, bile and protein solution samples. The assay was applied to pharmacokinetics of RDS, including plasma pharmacokinetics, excretion and protein binding studies. After i.v. 5, 20 and 100 mg x kg(-1) RDS via tail vein in rats, the plasma concentration-time profiles were fitted using 3P97 software. The average terminal half-life (t(1/2)) was 3.432 +/- 0.185 2 h. The relationship of dose and AUC of RDS was linear within the dosage range. This suggested that the disposition of RDS in rats belong to linear kinetics and the pharmacokinetic parameters of RDS were dose independent. After iv RDS 20 mg x kg(-1) in rats, the biliary excretion amount of parent drug amount was only 0.3181% +/- 0.2087% of given dosage, and the urinary excretion was 86.0% +/- 6.1% in 36 h. Ultrafiltration techniques were applied to determine the protein binding of RDS in plasma (from SD rat, Beagle dog and human), human serum albumin (HSA) and human alpha1-acid glycoprotein (AGP). The mean protein binding rate in plasma of SD rat, Beagle dog and human plasma of RDS were 80%-90%, in which the range of concentration of RDS was 5 to 100 microg x mL(-1). The protein binding to HSA was 85.7% +/- 1.3% and 14.0% +/- 3.2% to AGP.
Animals ; Area Under Curve ; Bile ; metabolism ; Dogs ; Half-Life ; Humans ; Injections, Intravenous ; Kinetics ; Male ; Orosomucoid ; metabolism ; Protein Binding ; Rats ; Rats, Sprague-Dawley ; Rutin ; administration & dosage ; analogs & derivatives ; blood ; pharmacokinetics ; urine ; Serum Albumin ; metabolism ; Solid Phase Extraction ; methods

OBJECTIVETo explore the effect of minimally invasive Ivor-Lewis esophagectomy on acute phase responses in patients with esophageal carcinoma.

METHODSForty-eight patients with middle or low thoracic esophageal carcinoma underwent Ivor-Lewis esophagectomy. The patients were divided into small incision group (n = 25) and conventional group (n = 23) according to the patients' will. Serum levels of acute phase proteins C reactive protein (CRP), haptoglobin (HPT), α₁-acid glycoprotein (α₁-AG), ceruloplasmin (CER), transferrin (TRF), β₂-microglobulin (β₂-MG), album protein (ALB), interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α) were measured and compared on 1st day before operation, at 18 hours as well as 3rd and 7th day after operation.

RESULTSThere was no significant difference in all the acute phase proteins indicators and IL-6 between the small incision and conventional groups at each time points after operation (P > 0.05). In both groups the levels of CRP, α₁-AG and HPT were significantly higher after operation than before operation (P < 0.05). The levels of ALB and TRF were significantly lower after operation than before operation (P < 0.05). The levels of CER and β₂-MG were not significantly different during perioperative period (P > 0.05). The level of TNF-α was significantly higher in the small incision group than that in the conventional group at the 18 hours postoperationally (P < 0.05), and were not significantly different on the other time points between the two groups (P > 0.05).

CONCLUSIONCompared with conventional operation, the small incision Ivor-Lewis esophagectomy do not significantly alleviate the stress of the surgical trauma in patients. Unchanging the essence of operation, if one is trying to minimize the stress caused by surgery on patients, the key factor is not the size of incision. An effective approach should be found in other operation-related factors.

Acute-Phase Proteins ; metabolism ; Aged ; C-Reactive Protein ; metabolism ; Carcinoma, Squamous Cell ; blood ; surgery ; Ceruloplasmin ; metabolism ; Esophageal Neoplasms ; blood ; surgery ; Esophagectomy ; methods ; Female ; Haptoglobins ; metabolism ; Humans ; Interleukin-6 ; blood ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Orosomucoid ; metabolism ; Perioperative Period ; Serum Albumin ; metabolism ; Serum Albumin, Human ; Transferrin ; metabolism ; Tumor Necrosis Factor-alpha ; blood ; beta 2-Microglobulin ; blood

OBJECTIVE: To screen potential plasma protein biomarkers for the progression of cervical precancerous lesions into cervical carcinoma and analyze their functions. METHODS: Plasma samples obtained from healthy control subjects, patients with low-grade squamous intraepithelial lesion (LSIL), high-grade squamous intraepithelial lesion (HSIL), cervical cancer (CC), and patients with CC after treatment were enriched for low-abundance proteins for liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. The MS data of the samples were analyzed using Discoverer 2.2 software, and the differential proteins (peptide coverage ≥20%, unique peptides≥2) were screened by comparison of LSIL, HSIL and CC groups against the control group followed by verification using target proteomics technology. Protein function enrichment and coexpression analyses were carried out to explore the role of the differentially expressed proteins as potential biomarkers and their pathological mechanisms. RESULTS: Compared with the control group, both LSIL group and HSIL group showed 9 differential proteins; 5 differentially expressed proteins were identified in CC group. The proteins ORM2 and HPR showed obvious differential expressions in LSIL and HSIL groups compared with the control group, and could serve as potential biomarkers for the progression of cervical carcinoma. The expression of F9 increased consistently with the lesion progression from LSIL to HSIL and CC, suggesting its value as a potential biomarker for the progression of cervical cancer. CFI and AFM protein levels were obviously decreased in treated patients with CC compared with the patients before treatment, indicating their predictive value for the therapeutic efficacy. Protein function enrichment analysis showed that all these differentially expressed proteins were associated with the complement system and the coagulation cascades pathway. CONCLUSIONS We identified 5 new protein biomarkers (F9, CFI, AFM, HPR, and ORM2) for cervical precancerous lesions and for prognostic evaluation of CC, and combined detection of these biomarkers may help in the evaluation of the development and progression of CC and also in improving the diagnostic sensitivity and specificity of cervical lesions.
Antigens, Neoplasm ; blood ; Biomarkers, Tumor ; blood ; Carrier Proteins ; blood ; Case-Control Studies ; Cervical Intraepithelial Neoplasia ; blood ; diagnosis ; Chromatography, Liquid ; Complement Factor I ; analysis ; Early Detection of Cancer ; Female ; Glycoproteins ; blood ; Haptoglobins ; Humans ; Neoplasm Proteins ; blood ; Orosomucoid ; analysis ; Precancerous Conditions ; blood ; diagnosis ; Serum Albumin, Human ; Tandem Mass Spectrometry ; Uterine Cervical Neoplasms ; blood ; diagnosis