The study aims to solve the instability problem of methylphenidate (MPH) in plasma, and establish a LC-MS/MS method for simultaneous determining of MPH in human plasma. The stabilities of MPH in different media were studied, and the degradation characteristics of MPH in these media were also investigated by HPLC and LC-MS/MS. To a 200 microL aliquot of freshly collected plasma sample, 10 microL 2% formic acid was added immediately to prevent the hydrolysis of MPH in human plasma samples. Chromatographic separation was performed on a Sapphire C18 column using the mobile phase of methanol - 5 mmol.L-1 ammonium acetate buffer solution containing 0.1% formic acid (46 : 54). MPH was quantified by tandem mass spectrometry operating in positive electrospray ionization mode with multiple reaction monitoring. The detection used the transitions of protonated molecules at m/z 234.2-->84.1 for MPH and m/z 260.3-->183.1 for propranolol (IS), separately. The intra- and inter-assay precisions were all below 5.0%. The accuracies were all in standard ranges. The linear calibration curve was obtained in the concentration range of 0.035-40 ng.mL-1. The methods fulfilled the demand. The method was used to determine the concentration of MPH in human plasma after a single dose of 36 mg MPH tablet to 6 healthy Chinese volunteers. The method is suitable for the precisely determination of MPH and for pharmacokinetic study of MPH in human plasma.

A sensitive method based on high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been developed for the simultaneous determination of folic acid (FA) and its active metabolite, 5-methyltetrahydrofolic acid (5-M-THF), in human plasma. The analytes were extracted from plasma with methanol solution containing 10 mg/mL of 2-mercaptoethanol and 0.025% (v/v) ammonium hydroxide. FA and 5-M-THF were more stable after the addition of 2-mercaptoethanol and ammonium hydroxide in the sample preparation procedures of this study than they were in the previously published methods. Chromatographic separation was performed on a Hedera ODS-2 column using a gradient elution system of acetonitrile and 1 mM ammonium acetate buffer solution containing 0.6% formic acid as mobile phase. LC-MS/MS was carried out with an ESI ion-source and operated in the multiple reaction monitoring (MRM) mode. The assay was linear over the concentration ranges of 0.249-19.9 ng/mL for FA, and 5.05-50.5 ng/mL for 5-M-THF. The developed LC-MS/MS method offers increased sensitivity for quantification of FA and 5-M-THF in human plasma and was applicable to a pharmacokinetic study of FA and 5-M-THF.

Objective:To consolidate ten years of clinical experience on the application of breast fiberoptic ductoscopy (FDS) in breast intra-ductal lesion. Methods:The clinical data of 1 368 cases of patients with nipple discharge were retrospectively collected and analyzed. Results:Significant differences were observed in the FDS diagnoses of patients with nipple discharge. The rates of tumor de-tection by FDS diagnosis were significantly higher when bloody and serous nipple discharge was used rather than milky and watery nip-ple discharge. For non-tumor nipple discharge, local drug perfusion via FDS was an effective treatment. A total of 303 patients had tu-mor resection or segmentectomy under localization via FDS, and 44 had segmentectomy after breast duct infusion of methylene blue. The diagnostic rate of localization via FDS (97.0%) was higher than that of breast duct infusion of methylene blue (86.4%). Conclu-sion:FDS is an accurate method for diagnosing patients with nipple discharge. In addition, it is also a good local drug perfusion method for patients with breast inflammatory nipple discharge ductoscopy. For patients with tumorous nipple discharge, localization via FDS can help improve the detection of the lesions, which can be removed by surgery.

OBJECTIVE:To establish a method for simultaneous determination of residual methylparaben,ethylparaben,nipa-sol and benzalkonium chloride in marketed eye drops. METHODS:HPLC method was adopted. The determination was performed on Hypersil GOLD C18 column with mobile phase consisted of 0.005 mol/L ammonium acetate(10 mL triehtylamine in 1 L solu-tion,pH adjusted to 5.0±0.5 with glacial acetic acid)-acetonitrile(45:55,V/V)at the flow rate of 1.0 mL/min. The detection wave-length was 262 nm(methylparaben,ethylparaben,nipasol)and 214 nm(benzalkonium chloride),respectively. The column tem-perature was 30 ℃ and sample size was 20 μL. RESULTS:The linear range were 1.2350-15.4380 μg/mL for methylparaben(r=0.9999),1.3170-16.3836 μ g/mL for ethylparaben (r=0.9997),1.2072-15.0894 μ g/mL for nipasol (r=0.9996) and 17.776-222.0 μg/mL for benzalkonium chloride(r=0.9999),respectively. Limits of quantitation were 2.0,2.0,2.0,1.11 μg,re-spectively;limits of determination were 0.375,0.375,0.375,0.333 μg,respectively. RSDs of precision,stability and reproducibili-ty tests were all lower than 2.0%. The average recoveries were 98.14%-102.48%(RSD=1.6%,n=9),98.79%-102.42%(RSD=1.3%,n=9),98.19%-102.49%(RSD=1.5%,n=9)and 98.76%-100.53%(RSD=0.6%,n=9),respectively. CONCLUSIONS:The method is accurate,reproducible,simple and suitable for the determination of residual methylparaben,ethylparaben,nipasol and benzalkonium chloride in marketed eye drops.

OBJECTIVETo investigate arsenic trioxide (As(2)O(3))-induced apoptosis and the effects on cell nuclear matrix related protein promyelocytic leukaemia (PML).

METHODSHepG2 cells were cultured in MEM medium and treated with 0.5, 2, 5 and 10 micro mol/L As(2)O(3) for either 24 h or 96 h at each concentration. In situ terminal deoxynucleotidyl transferase (TdT) labeling (TUNEL) and DNA ladders were used to detect apoptosis. Confocal microscopy and Western blotting were used to observe the expression of PML.

RESULTSThe growth rates of HepG2 cells were slower in the As(2)O(3) treated than the untreated control group. DNA ladder and TUNEL positive apoptotic cells could be detected in As(2)O(3) treated groups. The expression of PML decreased in HepG2 cells with 2 micro mol/L As(2)O(3) treatment. Confocal images demonstrated that the expression of PML protein in HepG2 cell nuclei decreased after treatment with 2 micro mol/L As(2)O(3), and micropunctates characteristic of PML protein in HepG2 cell nuclei disappeared after treatment with 5 micro mol/L As(2)O(3).

CONCLUSIONSOur results show that arsenic trioxide can significantly inhibit the growth of HepG2 cells in vitro. As(2)O(3) induces apoptosis in HepG2 tumor cells in a time and concentration dependent manner. As(2)O(3) may degrade the PML protein in HepG2 cell nuclei. The decreased expression of PML in As(2)O(3) treated tumor cells is most likely to be caused by apoptosis. Nuclear matrix associated protein PML could be the target of As(2)O(3) therapy.

Antineoplastic Agents ; pharmacology ; Arsenicals ; pharmacology ; Humans ; Neoplasm Proteins ; metabolism ; Nuclear Matrix ; drug effects ; metabolism ; Nuclear Proteins ; Oxides ; pharmacology ; Promyelocytic Leukemia Protein ; Transcription Factors ; metabolism ; Tumor Cells, Cultured ; Tumor Suppressor Proteins

Objective To compare the effect of two-dimensional strain imaging on evaluation of regional and global strain of right ventricle and hypertensive left ventriculaRhypertrophy in patients with hypertrophic cardiomyopathy.Methods A total of 100 cases were selected,in which 41 cases with hypertensive left ventriculaRhypertrophy were designed as observation group and 59 cases of hypertrophic cardiomyopathy were designed as study group.The left ventriculaRend diastolic diameter,left ventriculaRposterioRwall thickness,right ventriculaRfractional area change,right ventriculaRbasal dimension tracing,wall thickness of inter-ventriculaRseptum,tricuspid annulaRplane systolic execution,right ventriculaRregional and global longitudinal strain by two-dimensional strain imaging were compared between the two groups.Results The left ventriculaRend diastolic diameteRin the study group was significantly loweRthan the observation group (P<0.05),while the left ventriculaRposterioRwall thickness was significantly higheRthan the observation group (P<0.05).The right ventriculaRfractional area change and tricuspid annulaRplane systolic excution showed no significant difference between the two groups (P>0.05).The right ventriculaRbasal dimension tracing and wall thickness of inter-ventriculaRseptum in the study group were significantly loweRthan the observation group (P<0.05).The regional and global strain in the study group were significantly higheRthan the observation group (P<0.05).Conclusion Compared with hypertensive left ventriculaRhypertrophy,the two-dimensional strain imaging is more helpful foRindicating the presence of right ventriculaRfunction reconstructing in patients with hypertrophic cardiomyopathy.

Objective To compare the effect of two-dimensional strain imaging on evaluation of regional and global strain of right ventricle and hypertensive left ventriculaRhypertrophy in patients with hypertrophic cardiomyopathy.Methods A total of 100 cases were selected,in which 41 cases with hypertensive left ventriculaRhypertrophy were designed as observation group and 59 cases of hypertrophic cardiomyopathy were designed as study group.The left ventriculaRend diastolic diameter,left ventriculaRposterioRwall thickness,right ventriculaRfractional area change,right ventriculaRbasal dimension tracing,wall thickness of inter-ventriculaRseptum,tricuspid annulaRplane systolic execution,right ventriculaRregional and global longitudinal strain by two-dimensional strain imaging were compared between the two groups.Results The left ventriculaRend diastolic diameteRin the study group was significantly loweRthan the observation group (P<0.05),while the left ventriculaRposterioRwall thickness was significantly higheRthan the observation group (P<0.05).The right ventriculaRfractional area change and tricuspid annulaRplane systolic excution showed no significant difference between the two groups (P>0.05).The right ventriculaRbasal dimension tracing and wall thickness of inter-ventriculaRseptum in the study group were significantly loweRthan the observation group (P<0.05).The regional and global strain in the study group were significantly higheRthan the observation group (P<0.05).Conclusion Compared with hypertensive left ventriculaRhypertrophy,the two-dimensional strain imaging is more helpful foRindicating the presence of right ventriculaRfunction reconstructing in patients with hypertrophic cardiomyopathy.

Objective To investigate the effect of SABP, a water-soluble component of Salvia miltiorrhiza, on the growth of orthotopic transplantation of H22 liver cancer and the immune microenvironment of liver cancer. Methods We established a mouse model of orthotopic transplantation of H22 cell liver cancer in BALB/c mice. ELISA was used to detect the expression of PD-L1, TGF-β, IL-1β, IL-10, IL-4, IFN-γ, IL-18, IL-7, IL-2, CCL-2 and CCL-21 in the liver. We counted the organ indexes of liver, spleen and kidney. Results SABP inhibited the growth of orthotopic transplantation tumors of H22 cell liver cancer, and increased the expression levels of PD-L1, TGF-β, IL-1β and IL-10 in the microenvironment of liver cancer, as well as the liver, spleen and kidney coefficients. Conclusion SABP could inhibit the growth of orthotopic transplantation tumors of H22 cell liver cancer and promote the expression of PD-L1, TGF-β, IL-1β and IL-10 in the microenvironment of liver cancer.

We analyzed the characters of Sagliker syndrome by reporting a case of Sagliker syndrome and reviewed literature. We found that Sagliker syndrome had low incidence rate, young onset age, and was more common in women. There were high levels of alkaline phosphatase and parathyroid hormone in the blood of the patients. Patients with Sagliker syndrome with primary onset of non-diabetic nephropathy usually had chronic glomerulonephritis. We thought that secondary hyperparathyroidism in patients with Sagliker syndrome was induced by parathyroid hyperplasia, but high levels of alkaline phosphatase and parathyroid hormone in the blood of the patients with the secondary hyperparathyroidism were the main cause of Sagliker syndrome. Parathyroidectomy could stop the progress of Sagliker syndrome, but it could not reverse the occurrence of skeletal malformation.
Alkaline Phosphatase ; Chronic Disease ; Diabetic Nephropathies ; Female ; Humans ; Hyperparathyroidism, Secondary ; Parathyroidectomy ; Syndrome

Objective:To explore the effects of modified Duhuo Jisheng Decoction combined with Traditional Chinese Medicine (TCM) fumigation on joint function and inflammatory mediators in synovial fluid of patients with knee osteoarthritis.Methods:A total of 203 patients with knee osteoarthritis who met the inclusion criteria between March 2020 and March 2021 in the hospital were divided into observation group (102 cases) and control group (101 cases) according to the random number table method. The control group was given oral administration of celecoxib capsules + TCM fumigation, and the observation group was given modified Duhuo Jisheng Decoction on the basis of the control group. Both groups were treated for 3 months. TCM symptoms were scored before and after treatment, and Japanese Orthopaedic Association scores (JOA) was used to evaluate the knee function. Immunoturbidimetry was adopted to detect serum high-sensitivity C-reactive protein (hs-CRP), and ELISA was used for the detection of levels of TNF-α and IL-6 in synovial fluid, and the clinical efficacy was evaluated.Results:The total effective rate was 90.2% (92/102) in observation group and was 79.2% (80/101) in control group ( χ2=4.74, P=0.030). The JOA scores at 1 month and 3 months after treatment in observation group were significantly higher than those in the control group ( t=3.17, 7.74, all Ps<0.01). The scores of cold and painful knee joint, soreness and weakness of waist and knees, joint swelling and inhibited bending and stretching were significantly in the observation group after treatment lower than those in the control group ( t=7.61, 10.81, 6.62, 8.77, all Ps<0.001). The levels of TNF-α and IL-6 in synovial fluid and level of serum hs-CRP were significantly in the observation group after treatment lower than those in the control group ( t=7.97, 9.52, 9.56, all Ps<0.001). During treatment, the incidence rate of adverse reactions was 32.7% (33/101) in control group and that of observation group was 19.6% (20/102) ( χ2=4.49, P=0.034). Conclusion:Modified Duhuo Jisheng Decoction combined with TCM fumigation can improve the clinical symptoms and knee function, relieve the inflammatory response and enhance the clinical efficacy of patients with knee osteoarthritis.