[Hyperuricemia reduces semen parameters in infertile men].

OBJECTIVE: To explore the impact of hyperuricemia (HUA) on semen quality in infertile men. METHODS: Based on the level of fasting serum uric acid (SUA ≥420 µmol/L), 402 infertile men were divided into a normal SUA group (n = 304) and an HUA group (n = 98), and also into four age groups: 20－24, 25－29, 30－34, and 35－44 years old. Semen parameters were obtained from the patients by computer-assisted semen analysis and the levels of SUA determined by chemical colorimetry. RESULTS: The mean semen volume was significantly lower in the HUA than in the normal SUA group (2.40 vs 2.80 ml, P < 0.05), and so was the total sperm count (120.51 vs 187.21 ×106, P < 0.05). The mean semen volume was remarkably lower in the 25－29 and 30－34 years old patients with HUA than in those with normal SUA (2.40 and 2.55 ml vs 2.85 and 2.90 ml, P < 0.05), and so was the total sperm count in the 30－34 years old patients with HUA than in those with normal SUA (109.69 vs 192.91 ×106, P < 0.05). Correlation analysis showed that the level of SUA was negatively correlated with the semen volume (r = －0.193, P < 0.05) and total sperm count of the patients (r = －0.163, P < 0.05). CONCLUSIONS: Hyperuricemia significantly reduces the semen volume and total sperm count of infertile men, and the level of serum uric acid is closely related with semen quality.

[Correlation of semen parameters with inflammatory factors in the seminal plasma of obese males].

OBJECTIVE: To investigate the influence of inflammatory factors on semen parameters in the seminal plasma of obese men. METHODS: Based on the body mass index (BMI), 171 males were divided into a normal group (BMI < 24 kg/m2, n = 59), an overweight group (24 ≤ BMI < 28 kg/m2, n = 54), and an obesity group (BMI =≥ 28 kg/m2, n = 58). The routine semen parameters of the subjects were obtained by computer-assisted semen analysis, the levels of TNF-α, IL-6 and VEGF in the seminal plasma were measured by ELISA, and the correlation of BMI with the above indexes was analyzed. RESULTS: Sperm concentration was significantly decreased in the obesity group in comparison with the normal and overweight groups (ï¼»40.19 ± 24.05ï¼½ vs ï¼»66.54 ± 34.81ï¼½ and ï¼»57.73 ± 24.61ï¼½ ×106/ml, P <0.01), and so was the total number of sperm (ï¼»110.22 ± 75.44ï¼½ vs ï¼»200.75 ± 102.66ï¼½ and ï¼»157.46 ± 112.89ï¼½ ×106, P <0.01) and the percentage of progressively motile sperm (PMS) (ï¼»30.80 ± 15.56ï¼½ vs ï¼»50.75 ± 10.17ï¼½ and ï¼»39.71 ± 9.73ï¼½%, P <0.01). The levels of TNF-α and IL-6 in the seminal plasma were markedly elevated in the obesity group as compared with the normal and overweight groups (ï¼»76.90 ± 14.64ï¼½ vs ï¼»64.47 ± 11.92ï¼½ and ï¼»69.74 ± 12.32ï¼½ pg/ml, P <0.05; ï¼»54.17 ± 17.81ï¼½ vs ï¼»39.26 ± 9.09ï¼½ and ï¼»46.25 ± 13.66ï¼½ pg/ml, P <0.01), while that of VEGF remarkably reduced in the former group in comparison with the latter two (ï¼»154.24 ± 30.23ï¼½ vs ï¼»199.23 ± 36.28ï¼½ and ï¼»181.57 ± 34.41ï¼½ pg/ml, P <0.01). The levels of TNF-α, IL-6, and VEGF were significantly correlated with BMI (r = 0.254, 0.321 and －0.407, P <0.01), those of TNF-α and IL-6 negatively with the percentage of PMS (r =－0.163, P <0.05; r = －0.333, P <0.01). There was a positive correlation between TNF-α and IL-6 (r = 0.468, P <0.01), a negative correlation between IL-6 and VEGF (r = 0.177, P <0.05), but no correlation between TNF-α and VEGF (r = 0.058, P >0.05). CONCLUSIONS: The levels of TNF-α and IL-6 are increased and that of VEGF decreased in the seminal plasma of obese males, which may affect the semen quality.

Comparative proteomics of leaves found at different stem positions of maize seedlings.

To better understand the roles of leaves at different stem positions during plant development, we measured the physiological properties of leaves 1-4 on maize seedling stems, and performed a proteomics study to investigate the differences in protein expression in the four leaves using two-dimensional difference gel electrophoresis and tandem mass spectrometry in conjunction with database searching. A total of 167 significantly differentially expressed protein spots were found and identified. Of these, 35% are involved in photosynthesis. By further analysis of the data, we speculated that in leaf 1 the seedling has started to transition from a heterotroph to an autotroph, development of leaf 2 is the time at which the seedling fully transitions from a heterotroph to an autotroph, and leaf maturity was reached only with fully expanded leaves 3 and 4, although there were still some protein expression differences in the two leaves. These results suggest that the different leaves make different contributions to maize seedling growth via modulation of the expression of the photosynthetic proteins. Together, these results provide insight into the roles of the different maize leaves as the plant develops from a heterotroph to an autotroph.

Quantitative proteomics and transcriptomics reveal key metabolic processes associated with cotton fiber initiation.

An iTRAQ-based proteomics of ovules from the upland cotton species Gossypium hirsutum and its fuzzless-lintless mutant was performed, and finally 2729 proteins that preferentially accumulated at anthesis in wild-type ovules were identified. We confirmed that the gene expression levels of 2005 among these proteins also increased by performing an RNA sequencing transcriptomics. Expression of proteins involved in carboxylic acid metabolism, small-molecule metabolic processes, hormone regulation, and lipid metabolism was significantly enhanced in wild-type ovules. Quantitative real-time PCR verified the increased expression of 26 genes involved in these processes. Cotton 3-hydroxyacyl-CoA dehydratase (GhPAS2) catalyzing the third reaction of very long-chain fatty acid (VLCFA) biosynthesis, accumulated at anthesis in wild-type ovules. Heterogeneous expression of GhPAS2 restored viability to the Saccharomyces cerevisiae haploid psh1-deletion strain deficient in PAS2 activity. Application of VLCFA biosynthesis inhibitor acetochlor (2-chloro-N-[ethoxymethyl]-N-[2-ethyl-6-methyl-phenyl]-acetamide; ACE) and gibberellic acid to the unfertilized cotton ovules significantly suppressed fiber cell protrusion. In this study, the profiling of gene expression at both transcriptome and proteome levels provides new insights into cotton fiber cell initiation. BIOLOGICAL SIGNIFICANCE: Cotton fiber initiation determines the ultimate number of fibers per ovule, thereby determining fiber yield. In total, 2729 proteins were preferentially accumulated in wild-type ovules at anthesis. The most up-regulated proteins were assigned to carboxylic acid metabolism, small-molecule metabolic processes, hormone regulation, and lipid metabolism. In consistence with these findings, we characterized GhPAS2 gene coding for the enzyme that catalyzes VLCFA production. VLCFA biosynthesis inhibitor, acetochlor, was shown to significantly suppress fiber initiation. This study provides a genome-scale transcriptomic and proteomic characterization of fiber initial cells, laying a solid basis for further investigation of the molecular processes governing fiber cell development.

Molecular and biochemical characterization of a cyanogenic ß-glucosidase in the inner bark tissues of rubber tree (Hevea brasiliensis Muell. Arg.).

Tapping causes the loss of large amounts of latex from laticifers and subsequently enhances latex regeneration, a high carbon- and nitrogen-cost activity in rubber tree. It is suggested that a 67 kDa protein associated with protein-storing cells in the inner bark tissues of rubber tree plays an important role in meeting the nitrogen demand for latex regeneration. Here, the 67 kDa protein was further characterized by a combination of cell biological, molecular biological and biochemical techniques. Immunogold labeling showed that the 67 kDa protein was specifically localized in the central vacuole of protein-storing cells. A full-length cDNA, referred to as HbVSP1, was cloned. The HbVSP1 contained a 1584 bp open reading frame encoding a protein of 527 amino acids. The putative protein HbVSP1 shared high identity with the P66 protein from rubber tree and proteins of the linamarase, and bg1A from cassava (Manihot esculenta). HbVSP1 contained the active site sequences of ß-glucosidase, TFNEP and I/VTENG. In vitro analysis showed that the 67 kDa protein exhibited the activity of both ß-glucosidase and linamarase and was thus characterized as a cyanogenic ß-glucosidase. Proteins immuno-related to the 67 kDa protein were present in leaves and lutoids of laticifers. Tapping down-regulated the expression of HbVSP1, but up-regulated the expression of genes encoding the key enzymes for rubber biosynthesis, while the effect of resting from tapping was the reverse. Taken together, the results suggest that the 67 kDa protein is a vacuole-localized cyanogenic ß-glucosidase encoded by HbVSP1 and may have a role in nitrogen storage in inner bark tissues of trunk during the leafless periods when rubber tree is rested from tapping.

Chronic occupational exposure to hexavalent chromium causes DNA damage in electroplating workers.

BACKGROUND: Occupational exposure to chromium compounds may result in adverse health effects. This study aims to investigate whether low-level hexavalent chromium (Cr(VI)) exposure can cause DNA damage in electroplating workers. METHODS: 157 electroplating workers and 93 control subjects with no history of occupational exposure to chromium were recruited in Hangzhou, China. Chromium levels in erythrocytes were determined by graphite furnace atomic absorption spectrophotometer. DNA damage in peripheral lymphocytes was evaluated with the alkaline comet assay by three parameters: Olive tail moment, tail length and percent of DNA in the comet tail (tail DNA%). Urinary 8-OHdG levels were measured by ELISA. RESULTS: Chromium concentration in erythrocytes was about two times higher in electroplating workers (median: 4.41 µg/L) than that in control subjects (1.54 µg/L, P < 0.001). The medians (range) of Olive tail moment, tail length and tail DNA% in exposed workers were 1.13 (0.14-6.77), 11.17 (3.46-52.19) and 3.69 (0.65-16.20), and were significantly higher than those in control subjects (0.14 (0.01-0.39), 3.26 (3.00-4.00) and 0.69 (0.04-2.74), P < 0.001). Urinary 8-OHdG concentration was 13.65 (3.08-66.30) µg/g creatinine in exposed workers and 8.31 (2.94-30.83) µg/g creatinine in control subjects (P < 0.001). The differences of urinary 8-OHdG levels, Olive tail moment, tail length and tail DNA% between these two groups remained significant (P < 0.001) even after stratification by potential confounding factors such as age, gender, and smoking status. Chromium exposure was found to be positively associated with chromium levels in erythrocytes, urinary 8-OHdG levels, Olive tail moment, tail length and tail DNA%. Positive dose-response associations were also found between chromium levels in erythrocytes and Olive tail moment, tail length and tail DNA%. CONCLUSION: The findings in this study indicated that there was detectable chromium exposure in electroplating workers. Low-level occupational chromium exposure induced DNA damage.

[Clinical effect of Sizi Zhongwang capsule combined with Western medical therapy in treating male sterility].

OBJECTIVE: To observe the clinical effect of Sizi Zhongwang Capsule (SZC) combined with Western medicine (WM) in treating male sterility. METHODS: Two hundred and sixty-one male patients with sterility were assigned to 3 groups, 64 in the WM group were treated with conventional Western medical therapy alone, 87 in the SZC group were treated with SZC alone, and 110 in the combined group were treated with SZC combined Western medical therapy. The treatment lasted for 90 days in total. Changes of semen related parameters before and after treatment were observed, and the conditions of pregnancy in patients' spouse were followed-up. RESULTS: The difference in semen related parameters before and after treatment showed insignificant in the WM group (P > 0.05), but it did show statistical significance in the SZC group and the combined group (P < 0.05, P < 0.01). Moreover, the best effect was shown in the combined group, showing significant difference to the other two groups (P < 0.01, P < 0.05). The pregnancy rate of patients' spouse in the combined treated group was higher than that in the other two groups (P < 0.01, P < 0.05). CONCLUSION: SZC combined with Western medical therapy could effectively improve the quality of semen in males with infertility and enhance the pregnancy rate in their spouse.

Vegetative storage protein with trypsin inhibitor activity occurs in Sapindus mukorassi, a sapindaceae deciduous tree.

A vegetative storage protein (VSP) with trypsin inhibitor activity in a deciduous tree, Sapindus mukorassi, was characterized by means of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western-blot, immuno-histochemical localization, light- and electro-microscopy, together with analysis of proteinase inhibitor activity of the purified VSP in vitro. There were two proteins with molecular masses of about 23 and 27 kDa in a relatively high content in the bark tissues of terminal branches of S. mukorassi in leafless periods. The proteins decreased markedly during young shoot development, indicating their role in seasonal nitrogen storage. Immuno-histochemical localization with the polyclonal antibodies raised against the 23 kDa protein demonstrated that the 23 kDa protein was the major component of protein inclusions in protein-storing cells. The protein inclusions were identified by protein-specific staining and should correspond to the electron-dense materials in different forms in the vacuoles of phloem parenchyma cells and phloem ray parenchyma cells under an electron microscope. So, the 23 kDa protein was a typical VSP in S. mukorassi. The 23 and 27 kDa proteins shared no immuno-relatedness, whereas the 23 kDa protein was immuno-related with the 22 kDa VSP in lychee and possessed trypsin inhibitor activity. The 23 kDa protein may confer dual functions: nitrogen storage and defense.

[A simple protein in-gel digest method compatible with mass spectrometry analysis].

A simple protein in-gel digest method compatible with MALDI-TOF MS analysis was developed from the previously reported protocols with the following modifications. (1) The washing step was intensified by increasing the volume of distilled water and prolonging the vortex time. (2) Trypsin was acidified to improve the enzyme activity. (3) A Ca(2+) free pre-digest solution was used to reduce the trypsin auto cleavage. (4) The procedure of removal of salts and SDS was cancelled. (5) The digest was directly used to perform MALDI-TOF MS analysis. The compared results of this method with a latest reported protocol demonstrated that the method could efficiently reduce the peptide loss, provide more information for MS analysis, and consequently make the protein identification more reliable.

Vegetative storage protein in Litchi chinensis, a subtropical evergreen fruit tree, possesses trypsin inhibitor activity.

BACKGROUND AND AIMS: Vegetative storage proteins (VSPs) are commonly bioactive in herbaceous plants but few VSPs with bioactivity have been identified in trees. In addition, information on the characterization of VSPs in evergreen trees is limited. The objective of this study was to characterize the VSPs with bioactivity in evergreen trees. Methods The VSP in lychee (Litchi chinensis), an evergreen fruit tree, was characterized by a combination of cytological, biochemical and molecular biological techniques. KEY RESULTS: The VSP in lychee was a 22-kDa protein. It accumulated in the large central vacuoles of protein-storing cells (PSCs) in two distinguishable forms, granular and floccular. The PSCs were of a novel type. The 22-kDa protein is distributed in mature leaves, bark tissues of branches, trunk and large roots, paralleling the distribution of PSCs. Its homologues were present in mature seed. During young shoot development and fruiting, the 22-kDa protein decreased apparently, suggesting a nitrogen-storage function. The 22-kDa protein had several isoforms encoded by a small multigene family. One gene member, LcVSP1, was cloned. The LcVSP1 had no intron and contained a 675 bp open reading frame encoding a putative protein of 225 amino acids. LcVSP1 was homologous to Kunitz trypsin inhibitors. The 22-kDa protein inhibited trypsin and chymotrypsin, but had no inhibitory effect on subtilisin. CONCLUSIONS: Lychee is rich in a 22-kDa VSP with trypsin inhibitor activity. The VSP plays an important role in nitrogen storage while its possible defensive function remains to be elucidated.

[Clinical analysis of yuziwan in the treatment of 30 sterile men with abnormal protamine].

OBJECTIVE: To study the clinical application and effects of Yuziwan in the treatment of male sterility patient with abnormal protamine. METHODS: The changes of protamine, semen and sex hormones of 30 male sterility patients treated by Yuziwan were observed before and after the treatment. RESULTS: After a 3-month course of treatment, 9 cases were cured, 15 obviously improved and 6 failed to respond. The ratio of histone to protamine was decreased from (1.34 +/- 0.52) before the treatment to (0.72 +/- 0.32) after it, with significant difference (P < 0.01), the semen quality obviously improved (P < 0.05), and the LH and T levels markedly raised (P < 0.01). Yuziwan evidently improved the abnormal protamine, sperm quality and endocrine function of the sterility patients. CONCLUSION: Yuziwan has good curative effect on male sterility.