[Establishment of a method for detection of thiols in seminal plasma and its application in patients with hyperuricemia].

OBJECTIVE: To establish a method of reversed-phase ultra-performance liquid chromatography with fluorescence (UPLC-FL) detection for simultaneous determination of homocysteine (Hcy), cysteine (Cys), cysteine glycine (CysGly) and glutathione (GSH) and analysis of the contents of the four thiols in the seminal plasma of normal men and patients with hyperuricemia (HUA). METHODS: Seminal plasma samples were collected from 30 normal sperm donors and 30 HUA patients and reduced with tris (2-carboxyethyl) phosphine hydrochloride. Then, the samples were subjected to protein precipitation with trichloroacetic acid solution, derivative reaction with 7-fluorobenzofuran-4-sulfate and isolation with a C18 column, with 0.025 mol/L KH2PO4 (pH 3.0) for mobile phase A and pure methanol for B, followed by equal gradient elution with an excitation wavelength of 380 nm and an emission wavelength of 510 nm. RESULTS: The correlation coefficients (R2) of the four thiols all exceeded 0.999, with an average recovery rate of 94.23－107.87%. Compared with the normal sperm donors, the HUA patients showed significantly increased contents of Cys (ï¼»108.01 ± 48.03ï¼½ vs ï¼»250.10 ± 55.87ï¼½ µmol/L, P < 0.05), Hcy (ï¼»113.97 ± 6.32ï¼½ vs ï¼»48.35 ± 15.07ï¼½ µmol/L, P < 0.05), and GSH (ï¼»3.15 ± 1.48ï¼½ vs ï¼»4.63 ± 1.17ï¼½ µmol/L, P < 0.05), but a decreased level of CysGly (ï¼»12.79 ± 3.18ï¼½ vs ï¼»5.94 ± 0.99ï¼½ µmol/L, P < 0.05). CONCLUSIONS: The method of reversed-phase UPLC-FL detection established in this study has made it possible simultaneous detection of Hcy, Cys, CysGly and GSH in the seminal plasma, which is applicable to laboratory research and clinical routine examination. Patients with hyperuricemia may incur oxidative damage to the reproductive system.

[Ultra-performance liquid chromatography for determination of the contents of biogenic amines in the seminal plasma].

OBJECTIVE: To establish an ultra-performance liquid chromatography (UPLC) method for determination of the contents of the three biogenic amines putacine, spermidine and spermine in human seminal plasma. METHODS: Seminal plasma samples were extracted with 5% trichloroacetic acid and processed by pre-column derivatization with dansyl chloride. Chromatographic separation was performed with a C18 (2.1×50 mm,1.7 µm) chromatographic column using water and acetonitrile for mobile-phase gradient elution at a flow rate of 0.3 ml/min, a detection wavelength of 245 nm, a column temperature of 35℃ and an injection volume was 3.0 µl. The contents of putacine, spermidine and spermine in the seminal plasma of 52 healthy sperm donors (the normal group) and 23 azoospermia patients (the AS group) were measured, and their correlation with routine semen parameters were analyzed. RESULTS: The three biogenic amines showed a good linearity (r ≥ 0.999), with a lower detection limit of 0.03－0.08 µg/ml. The relative standard deviation (RSD) of precision was ≤ 0.72% and the average recovery rate was 79.74%－108.87%. The normal group, compared with the AS patients, showed significantly higher contents of putrescine (ï¼»8.19 ± 7.85ï¼½ vs ï¼»2.43 ± 1.38ï¼½ mg/ml, P < 0.05), spermidine (ï¼»77.30 ± 32.58ï¼½ vs ï¼»31.99 ± 16.21ï¼½ mg/ml, P < 0.05) and spermine (ï¼»246.44 ± 83.99ï¼½ vs ï¼»166.15 ± 79.28ï¼½ mg/ml, P < 0.05). However, the contents of the three biogenic amines in the seminal plasma exhibited no significant correlation with the routine semen parameters in the normal group. CONCLUSIONS: The ultra-performance liquid chromatography method we established, with the advantages of high sensitivity and reproducibility and short peak-time, can quickly and accurately determine the contents of biogenic amines in the seminal plasma.

[Correlation of seminal plasma biochemical indexes with routine semen parameters in sperm donors].

OBJECTIVE: To investigate the relationship of seminal plasma biochemical indexes with routine semen parameters and that between seminal plasma biochemical indexes. METHODS: Using the automatic biochemical analyzer, we measured the contents of neutral α-glucosidase (NAG), fructose hexokinase (Fru) , citric acid (CA), acid phosphatase (ACP), (zinc) Zn, uric acid (UA), lactase dehydrogenase (LDH) and α-hydroxybutyrate dehydrogenase (α-HBDH) in the seminal plasma of 84 sperm donors in the Human Sperm Bank of Hebei Province. We analyzed the correlation between these indexes and that between routine semen parameters and these indexes. RESULTS: Sperm concentration and total sperm count were correlated positively with the contents of seminal plasma NAG, ACP, Zn, CA, LDH and α-HBDH (P<0.05) but negatively with Fru (P<0.05), the percentage of progressively motile sperm positively with seminal plasma Zn (P<0.05), and CA positively with NAG, Zn, LDH, α-HBDH and ACP (P<0.01) but negatively with Fru (P<0.01), NAG positively with Zn, LDH, α-HBDH and ACP (P<0.05) but Fru negatively with ACP (P<0.01), Zn positively with LDH, α-HBDH and ACP (P<0.01), and LDH positively with α-HBDH and ACP (P<0.01) but UA negatively with ACP (P<0.05). CONCLUSIONS: Biochemical indexes in the seminal plasma of healthy men are not only closely related to each other, but also to some routine semen parameters.

[Clinical progress of paroxysmal nocturnal hemoglobinuria].

Through the applications of high-sensitivity flow cytometry of FLAER and the treatment of eculizumab, it is necessary to understand of paroxysmal nocturnal hemoglobinuria (PNH) from a new point of view. The results of studies demonstrate that treatment with eculizumab alters the natural history of PNH by virtually eradicating thromboembolic complications, inhibiting of intravascular hemolysis and reducing or eliminating transfusion requirements. Eculizumab treatment may also reduce disease-related mortality. This review focuses on the studies to define the relationship between PNH and bone marrow failure syndromes and to characterize the long-term outcome of patients with PNH treated with eculizumab. New therapeutic strategies aimed at controlling extravascular and intravascular hemolysis are discussed.