Enhancement of Frozen-Thawed Human Sperm Quality with Zinc as a Cryoprotective Additive.

BACKGROUND Cryopreservation preserves male fertility, crucial in oncology, advanced age, and infertility. However, it damages sperm motility, membrane, and DNA. Zinc (Zn), an antioxidant, shows promise in improving sperm quality after thawing, highlighting its potential as a cryoprotectant in reproductive medicine. MATERIAL AND METHODS Gradient concentration of ZnSO4 (0, 12.5, 25, 50, and 100 µM) was added in the Glycerol-egg yolk-citrate (GEYC) cryopreservative medium as an extender. Alterations in sperm viability and motility parameters after cryopreservation were detected in each group. Sperm plasma membrane integrity (PMI), acrosome integrity (ACR), DNA fragment index (DFI), and changes in sperm mitochondrial function were examined, including: mitochondrial potential (MMP), sperm reactive oxygen species (ROS), and sperm ATP. RESULTS We found that 50 µM ZnSO4 was the most effective for the curvilinear velocity (VCL) and the average path velocity (VAP) of sperm after cryo-resuscitation. Compared to the Zn-free group, sperm plasma membrane integrity (PMI) was increased, DNA fragmentation index (DFI) was decreased, reactive oxygen species (ROS) was reduced, and mitochondrial membrane potential (MMP) was increased after cryorevival in the presence of 50 µM ZnSO4. CONCLUSIONS Zn ion is one of the antioxidants in the cell. The results of our current clinical study are sufficient to demonstrate that Zn can improve preserves sperm quality during cryopreservation when added to GEYC. The addition of 50 µM ZnSO4 increased curve velocity, mean path velocity, sperm survival (or plasma membrane integrity), and mitochondrial membrane potential while reducing ROS production and DNA breaks compared to GEYC thawed without ZnSO4.

Association of SNPs in zinc transporter genes on seminal plasma zinc levels in humans.

This study is to examine the effects of single nucleotide polymorphisms (SNPs) of SLC30A and SLC39A on seminal plasma zinc concentration. Blood and seminal plasma samples were collected from outpatients. SNPs of zinc transporters were analyzed by next Generation sequencing technology, and seminal plasma zinc concentration were determined by inductively coupled plasma optical emission spectrometry. Our date showed nine SNPs (SLC30A8 rs2466295, rs2466294, SLC30A10 c.-160 C>G, SLC39A8 rs9331, rs9705, rs151392, rs151393, SLC39A11 rs9912126, SLC39A14 rs1051708) were significantly associated with seminal plasma zinc concentration, and 14 SNPs (SLC30A8 rs2466295, rs2466294, SLC30A10 c.-160 C>G, SLC39A6 rs148550301, SLC39A8 rs9331, rs9705, rs151392, rs151393, SLC39A11 rs9912126, rs61736066, rs36041371 and SLC39A14 rs1051708, rs76963096, rs17060854) were found to be significantly associated with total zinc per ejaculate. The seminal plasma zinc concentrations and total zinc per ejaculate were associated with the number of SNPs, and decreased significantly when five SNPs (SLC39A8 rs9331, rs9705, rs151392, rs151393 and SLC39A14 rs1051708) were a combination of homozygous genotype. Our findings suggest that different zinc transporter SNPs may significantly affect seminal plasma zinc levels.

Differentially expressed microRNA in testicular tissues of hyperuricaemia rats.

This study is to identify the differentially expressed miRNAs in testicular tissues of rats with hyperuricaemia-induced male infertility. We found that the hyperuricaemia model group had significantly increased serum uric acid, while significantly decreased sperm concentration and motile sperm percentage than normal group (p < .05). A total of 39 differentially expressed miRNAs were identified in the testicular tissues of hyperuricaemia rats compared with the control rats, ten of which were validated by real-time PCR. The target mRNAs of 7 differentially expressed miRNAs (miR-10b-5p, miR-26a-5p, miR-136-5p, miR-151-3p, miR-183-5p, miR-362-3p and miR-509-5p) from 3'-untranslated region binding perspective were enriched in signalling pathways of Wnt, Jak-STAT, mTOR and MAPK. The target mRNAs of 6 differentially expressed miRNAs (miR-136-5p, miR-144-3p, miR-99a-5p, miR-509-5p, miR-451-5p and miR-362-3p) from coding sequence binding perspective were enriched in signalling pathways of Calcium, Notch and MAPK. The functions of miRNAs in testicular tissues of rats with hyperuricaemia were revealed by the differentially expressed miRNAs (miR-183-5p, miR-99a-5p, miR-10b-5p, miR-151-3p, miR-26a-5p, miR-451-5p, miR-362-3p, miR-136-5p, miR-144-3p and miR-509-5p)-mRNAs interaction network. The differentially expressed miRNAs in the testicular tissues of hyperuricaemia rats might shed light on the mechanism of hyperuricaemia-induced male infertility.

[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.

[Correlation of seminal plasma zinc alpha-2 glycoprotein with semen quality in obese males].

OBJECTIVE: To investigate the relationship between seminal plasma zinc alpha-2 glycoprotein (ZAG) and semen quality in obese males. METHODS: This study included 130 obese male patients with idiopathic infertility Based on the concentration of seminal plasma ZAG, we divided the patients into three tertile groups: tertile 1 (T1, 73.45－97.15 µg/ml, n = 43), T2 (97.16－115.46 µg/ml, n = 44), and T3 (115.47－220.11 µg/ml, n = 43). We measured the concentrations of seminal plasma zinc (SPZ) and ZAG of the patients by ELISA, obtained the semen parameters, and analyzed the correlation of semen quality with the levels of SPZ and ZAG and the influence of obesity on SPZ, ZAG and semen quality. RESULTS: The mean level of seminal plasma ZAG in the 130 obese male patients was (111.29 ± 26.50) µg/ml. There were statistically significant differences in sperm concentration and total sperm count among the three tertile groups (P < 0.05). The level of seminal plasma ZAG was correlated negatively with the body mass index (BMI), waist circumference (WC), sperm concentration and sperm count (P < 0.01), that of SPZ positively with BMI and WC (P < 0.05) but negatively with semen volume and the percentage of progressively motile sperm (P < 0.05). The level of serum ZAG, however, exhibited no correlation with SPZ, seminal plasma ZAG or semen quality. Obesity was found to be associated with significantly decreased concentration of seminal plasma ZAG and percentage of progressively motile sperm but remarkably increased level of SPZ (P < 0.05). CONCLUSIONS: Obesity may induce the metabolic disorder of SPZ and ZAG, change the microenvironment of seminal plasma, and consequently affect semen quality.

[Correlation of the levels of seminal plasma homocysteine, folate and cobalamin with semen parameters in obese men].

OBJECTIVE: To investigate the correlation of seminal plasma homocysteine (Hcy) and its metabolic factors folate (FA) and cobalamin (VB12) with semen quality in obese men. METHODS: We randomly selected 83 male patients with idiopathic infertility for this study and, according to the body mass index (BMI), divided them into a normal BMI (n = 28), an overweight (n = 28) and an obesity group (n = 27). We determined the levels of Hcy, FA and VB12 in the seminal plasma by ELISA and analyzed their correlation with the semen parameters of the patients in different groups. RESULTS: Compared with the normal BMI group, the obese males showed significant decreases in the semen volume (ï¼»3.23 ± 0.86ï¼½ vs ï¼»2.58 ± 0.77ï¼½ ml, P < 0.05), total sperm count (ï¼»191.35 ± 103.00ï¼½ vs ï¼»121.55 ± 88.08ï¼½ ×106, P < 0.05), percentage of progressively motile sperm (ï¼»52.88 ± 15.58ï¼½ % vs ï¼»38.97 ± 16.52ï¼½ %, P < 0.05), seminal plasma Hcy (ï¼»7.41 ± 1.29ï¼½ vs ï¼»6.62 ± 0.85ï¼½ µmol/L, P < 0.05), and VB12 (ï¼»282.41 ± 30.38ï¼½ vs ï¼»230.07 ± 37.75ï¼½ pmol/L, P < 0.05). There were no statistically significant differences in the semen parameters between the overweight group and the normal BMI or the obese group (P > 0.05), or in sperm concentration or the FA level among the three groups of patients (P > 0.05). The levels of seminal plasma Hcy and VB12 were correlated positively with the semen volume (r = 0.281 and 0.242, P < 0.05) and total sperm count (r = 0.229 and 0.258, P < 0.05) but negatively with BMI (r = －0.293 and －0.238, P < 0.05). No correlation, however, was found either between sperm concentration and the percentage of progressively motile sperm (P > 0.05) or between the FA level and BMI or semen parameters (P > 0.05). CONCLUSIONS: The levels of seminal plasma Hcy and VB12 are correlated with BMI, semen volume and total sperm count, which suggests that the concentrations of seminal plasma Hcy and VB12 may be associated with the fertility of obese men.

[Correlation of insulin resistance with male reproductive hormone levels and semen parameters].

OBJECTIVE: To investigate the influence of insulin resistance on male reproductive hormones and semen quality. METHODS: Using the electrochemiluminescence method, we measured the levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), estradiol (E2) and testosterone (T) in the serum of 83 infertile males. We detected the levels of fasting plasma glucose (FPG) and fasting insulin (FINS) and calculated the insulin resistance index presented as homeostasis model assessment of insulin resistance (HOMA-IR). Based on HOMA-IR, we divided the patients into three tertile groups, T1 (HOMA-IR 0.36－0.55, n = 27), T2 (HOMA-IR 0.56－0.80, n = 28) and T3 (HOMA-IR 0.81－1.97, n = 28), obtained their semen parameters by computer-assisted semen analysis (CASA) and analyzed the correlation of HOMA-IR with male reproductive hormone levels and semen parameters. RESULTS: With the elevation of HOMA-IR, the patients of the T1, T2 and T3 groups showed significant decreases in the serum T level (ï¼»14,26 ± 4.27ï¼½ vs ï¼»14.75 ± 5.00ï¼½ vs ï¼»11.62 ± 3.68ï¼½ nmol/L, P <0.05) and the percentage of progressively motile sperm (PMS) (ï¼»51.04 ± 15.10ï¼½% vs ï¼»48.04 ± 16.24ï¼½% vs ï¼»37.84 ± 18.23ï¼½%, P <0.05). HOMA-IR was correlated negatively with the serum T level (r = －0.333, P = 0.002), semen volume (r = －0.23, P = 0.029) and PMS (r = －0.27, P = 0.015), and so was FINS with the serum T level (r = －0.327, P = 0.003) and PMS (r = －0.315, P = 0.004), while the semen volume was correlated positively with the levels of serum T (r = 0.221, P = 0.048) and FSH (r = 0.222, P = 0.047). Multivariate linear regression analysis showed that HOMA-IR was an independent influencing factor for PMS and the body mass index (BMI) was that for the semen volume and total sperm count. CONCLUSIONS: Insulin resistance may reduce semen quality by changing the levels of male reproductive hormones.

[Correlation of reproductive hormone levels and seminal plasma oxidative stress with semen quality in obese males].

OBJECTIVE: To investigate the correlation of the levels of reproductive hormones and oxidative stress in the seminal plasma with semen parameters in obese males. METHODS: Based on the body mass index (BMI), we divided 138 infertile men into three groups: normal (BMI <24 kg/m2, n = 48), overweight (24 kg/m2≤BMI<28 kg/m2, n = 47), and obesity (BMI ≥28 kg/m2, n = 43). We determined the concentrations of follicle-stimulating hormone (FSH), luteotropic hormone (LH), prolactin (PRL), testosterone (T) and estradiol (E2) in the serum by electrochemiluminescence and measured the levels of superoxide dismutase (SOD), glutathione-S-transferases (GSTs), reactive oxygen species (ROS) and malondialdehyde (MDA) in the seminal plasma by ELISA, compared the above indexes among the three groups, and analyzed their correlation with the semen volume, sperm concentration, total sperm count, and percentage of progressively motile sperm (PMS). RESULTS: The semen volume was significantly lower in the obesity than in the normal group (ï¼»2.63 ± 0.74ï¼½ vs ï¼»3.37 ± 1.00ï¼½ ml, P < 0.05), and so was the percentage of PMS in the overweight and even lower in the obesity than in the normal group (ï¼»47.91 ± 12.89ï¼½ and ï¼»41.27 ± 15.77ï¼½ vs ï¼»54.04 ± 13.29ï¼½%, P < 0.05). Compared with the normal group, both the overweight and obesity groups showed markedly decreased levels of serum T (ï¼»4.83 ± 1.42ï¼½ vs ï¼»3.71 ± 1.22ï¼½ and ï¼»3.49 ± 1.12ï¼½ ng/ml, P<0.05), T/LH ratio (1.53 ± 0.57 vs 1.19 ± 0.54 and 0.97 ± 0.51, P<0.05), SOD (ï¼»112.05 ± 10.54ï¼½ vs ï¼»105.85 ± 6.93ï¼½ and ï¼»99.33 ± 8.39ï¼½ U/ml, P<0.05), and GSTs (ï¼»31.75±6.03ï¼½ vs ï¼»29.54±5.78ï¼½ and ï¼»29.02±4.52ï¼½ U/L, P<0.05), but remarkably increased seminal plasma ROS (ï¼»549.93±82.41ï¼½ vs ï¼»620.61±96.13ï¼½ and ï¼»701.47±110.60ï¼½ IU/ml, P<0.05) and MDA (ï¼»7.46 ± 2.13ï¼½ vs ï¼»8.72 ± 1.89ï¼½ and ï¼»10.47 ± 2.10ï¼½ nmol/L, P<0.05). BMI was correlated positively with ROS and MDA, but negatively with the semen volume, PMS, T, T/LH, SOD and GSTs (P<0.05); LH negatively with sperm concentration, total sperm count and GSTs (P<0.05); PRL negatively GSTs (P<0.05); E2 positively with SOD (P<0.05); T positively with SOD (P<0.05) but negatively with MDA (P<0.05); T/LH positively with PMS and SOD (P<0.05) but negatively with ROS and MDA (P<0.05); SOD positively with semen volume, PMS and GSTs (P<0.05) but negatively with ROS and MDA (P<0.05); GSTs negatively with sperm concentration; total sperm count and MDA (P<0.05); ROS positively with MDA (P<0.01) but negatively with PMS (P<0.05); and MDA negatively with semen volume (P<0.05). Multivariate logistic regression analysis showed that the independent factors influencing the semen volume were BMI and GSTs, those influencing the total sperm count were BMI and T, and those influencing PMS were BMI and MDA. CONCLUSIONS: Increased BMI induces changes in the levels of male reproductive hormones and seminal plasma oxidative stress and affects semen quality, which may be associated with male infertility.

[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 serum zinc alpha 2 glycoprotein with blood lipid and reproductive hormone levels in men].

OBJECTIVE: To study the changes of the serum zinc alpha 2 glycoprotein (ZAG) level in men and its relationship with blood lipid male reproductive hormones. METHODS: We enrolled 297 men aged 25－ 65 years in this study, 152 with hyperlipemia (HL) and the other 145 with normal blood lipid (normal control). We divided them into four age groups (25－35 yr, 36－45 yr, 46－55 yr, and 56－65 yr) and three tertile groups (Q1, Q2, and Q3) according to the tertiles of the serum ZAG level, and examined their blood lipid, blood glucose, serum ZAG, and reproductive hormones. RESULTS: The serum ZAG level was decreased gradually with the increase of age in both the HL patients and normal controls, significantly in the 36－45 and 56－65 yr age groups (P <0.05), and markedly lower in the HL than in the control men in the 25－35 and 36－45 yr groups (P <0.05). The levels of follicle-stimulating hormone (FSH) and total testosterone (TT) changed significantly with the ZAG level. The level of serum ZAG was correlated negatively with age (r = －0.58, P<0.05), waist circumference (r = －0.21, P <0.05), body mass index (BMI) (r = －0.22, P <0.05), fasting blood glucose (r = －0.16, P <0.05) , and triglyceride (TG) (r = －0.27, P <0.05) but positively with TT (r = 0.36, P <0.05). Age, BMI and TG were independent factors influencing the serum ZAG level. CONCLUSIONS: The serum ZAG level is decreased with the increase of age and associated with lipid metabolism, abdominal obesity, and reproductive hormone levels in males.

[Metabonomics in the diagnosis and treatment of male infertility].

Metabonomics is an emerging branch of science for the study of endogenous small molecule metabolites in organisms, which plays an important role in evaluatingthediagnosis and treatment of male infertility by exploring the metabolites of body fluids, cells and tissues. With its advantages ofmass information, noninvasiveness, and celerity, metabonomics will be widely applied to clinical researches in the future. This review introducesmetabonomics and its analytical techniques and data processing procedures,its latest application in the studies of the etiology, diagnosis and the treatment of male infertility, and the prospect of its future application in the researches of male reproduction.

[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.

[Analysis of serum reproductive hormone levels in men with hyperlipemia in the urban area of Shijiazhuang].

OBJECTIVE: To investigate the aging-related changes of reproductive hormones in hyperlipemia men in the urban area of Shijiazhuang and the relationship between the levels of reproductive hormones and that of blood lipid. METHODS: We enrolled 297 males aged 25－65 years in this study, determined the levels of serum total testosterone (TT), luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol (E2), prolactin (PRL), triglyeride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and fasting blood glucose (FBG), and compared the parameters among different age groups. RESULTS: The level of serum TT was decreased while that of FSH increased significantly in the 36－45 years group and that of LH elevated only in the 56－65 years old hyperlipemia males(P<0.05). No statistically significant changes were observed in PRL and E2 among different age groups. Spearman correlation analysis manifested that age was correlated negatively with TT, FSH, and LH (r=－0.266, 0.514, and 0.350, P<0.05), TT weakly negatively with TC and TG (r=－0.215 and －0.286, P<0.05) but weakly positively with HDL-C (r=0.281, P<0.01), and FSH and LH weakly positively with TG (r=0.282 and 0.343, P<0.05). Regression analysis showed age to be an independent influencing factor of TT, FSH and LH, while HDL-C to be that of TT. CONCLUSIONS: The levels of serum TT and FSH gradually change with aging and low TT and high FSH and LH are closely related to hyperlipemia in adult men.

Biochemical Markers of Zinc Nutrition.

Zinc is an important trace element involved in the biochemical and physiological functions of the organism and is essential in the human body. It has been reported that 17.3% of people around the world are at risk of many diseases due to zinc deficiency, which has already affected people's healthy lives. Currently, mild zinc deficiency is difficult to diagnose early due to the lack of typical clinical manifestations, so finding zinc biomarkers is crucial for people's health. The present article reviews the main representative zinc biomarkers, such as body fluid zinc levels, zinc-dependent proteins, tissue zinc, and zinc-containing enzymes, to provide a reference for actively promoting the study of zinc nutritional status and early clinical diagnosis.

Zinc Ameliorates Tripterygium Glycosides-Induced Reproductive Impairment in Male Rats by Regulating Zinc Homeostasis and Expression of Oxidative Stress-Related Genes.

Tripterygium glycosides (TG) can seriously damage male reproductive function, and the reproductive system is difficult to restore after stopping the administration of TG in male rats. Zinc (Zn) is one of the most important trace elements in the human body and plays an important role in maintaining male fertility. The aim of this study was to investigate whether zinc supplementation could improve the testicular reproductive damage induced by TG toxicity in rats and to investigate its mechanism of action. The results showed that zinc sulfate (ZnSO4) could improve testicular tissue structure and semen parameters, promote testosterone synthesis, increase zinc-containing enzyme activity, increase zinc concentration in serum and testicular tissues, and maintain zinc homeostasis in male rats induced by TG toxicity. Zinc supplementation activated relevant signalling molecules in the KEAP1-NRF2/ARE pathway and alleviated TG-induced oxidative stress. Therefore, this study concluded that zinc supplementation could improve reproductive damage by regulating zinc homeostasis and the expression of genes related to oxidative stress.

Palliative Effect of Combined Application of Zinc and Selenium on Reproductive Injury Induced by Tripterygium Glycosides in Male Rats.

The long-term use of tripterygium glycosides (TG) can lead to male reproductive damage. Research indicates that zinc and selenium exhibit a synergistic effect in the male reproductive system, with the combined preparation demonstrating superior therapeutic effects compared to individual preparations. The purpose of this study was to explore the specific mechanism by which zinc and selenium mitigate reproductive toxicity induced by TG in male rats. Rats were randomly assigned to three groups: control group (C group), model group (M group, receiving TG at 30 mg/kg/day), and model + zinc + selenium group (ZS group). The ZS group was also given TG gavage for the first 4 weeks. Starting from the fifth week until the conclusion of the eighth week, the ZS group received an additional protective treatment of 10 mg/kg/day Zn and 0.1 mg/kg/day Se 4 h after TG administration. Following euthanasia, blood samples, rat testis, and epididymis tissues were collected for further experiments. Combined zinc-selenium treatment corrects the imbalance of zinc-selenium homeostasis in testicular tissue induced by TG. This is achieved by upregulating the expression of metal transcription factor (MTF1) and zinc transporters ZIP8 and ZIP14 and downregulating the expression of ZnT10. Improvement of zinc and selenium homeostasis enhanced the expression of zinc-containing enzymes (ADH, LDH, and ALP) and selenoproteins (GPx1 and SELENOP) in the testis. At the same time, zinc and selenium mitigate TG-induced reproductive damage by promoting the activity of antioxidant enzymes and upregulating the expression of proteins associated with the oxidative stress pathway, including Nrf2, Keap1, HO-1, PI3K, and p-AKT.

A Dietary Supplement Jinghuosu Ameliorates Reproductive Damage Induced by Tripterygium Glycosides.

OBJECTIVE: To determine the possible protective effects of Jinghuosu, a dietary supplement (DS), on tripterygium glycosides (TG)-induced reproductive system injury in rats and its underlying mechanisms. METHODS: A reproductive damage model was established in rats by feeding of TGs. Twenty-eight male Sprague Dawley rats were randomly divided into 4 groups using a random number table (n=7 in each): control (C) group, model (M) group, DS group and L-carnitine (LC) group. Rats in M, DS and LC groups received 40 mg/kg TGs orally. Starting from the 5th week, after administration of TGs for 4 h every day, rats in DS and LC groups were administered with 2.7 g/kg DS and 0.21 g/kg LC, respectively, for protective treatment over the next 4 weeks. Rats in Group C continued to receive the control treatment. Hematoxylin-eosin staining was used for histopathological analysis of rat testicular tissues. Enzyme-linked immunosorbent assay was performed to measure alkaline phosphatase (ALP), lactate dehydrogenase, alcohol dehydrogenase, total antioxidant capacity (T-AOC), superoxide dismutase, glutathione peroxidase (GSH-Px), and malondialdehyde (MDA) concentrations. Chemiluminescence assay was used to determine the serum testosterone content. Quantitative real-time PCR and Western blotting were conducted to analyze the expression of genes and proteins related to the testosterone synthesis pathway and the nuclear factor erythroid 2-related factor 2/heme oxygenase 1 antioxidant pathway. RESULTS: Oral administration of TGs induced significant increases in the testicular levels of zinc transporter 1 and MDA (P<0.05). On the other hand, sperm concentration, sperm motility, and serum testosterone, serum zinc, testicular zinc, Zrt-, Irt-like protein 1, ALP, luteinizing hormone (LH) receptor, steroidogenic acute regulatory protein, Cytochrome P450 family 11 subfamily A member 1, 3 ß -hydroxysteroid dehydrogenase 1 T-AOC, GSH-Px, nuclear factor erythroid 2-related factor 2, heme oxygenase-1 and NAD (P)H: quinone oxidoreductase 1 levels decreased following TGs exposure (P<0.05). All of these phenotypes were evidently reversed by DS (P<0.05). CONCLUSION: DS Jinghuosu protects against TG-induced reproductive system injury in rats, probably by improving zinc homeostasis, enhancing the testosterone synthesis and attenuating oxidative stress.

Effects of Zinc Combined with Metformin on Zinc Homeostasis, Blood-Epididymal Barrier, and Epididymal Absorption in Male Diabetic Mice.

Diabetes increases the likelihood of germ cell damage, hypogonadism, and male infertility. Diabetes leads to lower zinc (Zn) levels, an important micronutrient for maintaining male fertility, and zinc deficiency can lead to decreased male fertility through multiple mechanisms. The aim of this study was to investigate the effect of combined metformin and zinc administration on epididymis in diabetic mice; 10 of 50 male mice were randomly selected as the control group (group C), and the remaining 40 mice were randomly divided into untreated diabetes group (group D), diabetes + zinc group (group Z), diabetes + metformin group (group M), and diabetes + metformin + zinc group (group ZM) with 10 mice each. Diabetic mice in group Z received oral zinc (10 mg/kg) once daily for 4 weeks; diabetic mice in group M received oral metformin (200 mg/kg) once daily for 4 weeks; diabetic mice in group ZM received oral metformin and zinc once daily for 4 weeks; and groups C and D received the same amount of sterile water by gavage. Overnight fasted mice were sacrificed, and blood samples, mouse epididymides, and sperm were collected for further experiments. In group D, fasting blood glucose and insulin resistance index increased significantly, semen quality, serum insulin, and testosterone decreased, and epididymal structure was disordered. In group D, epididymal tissue zinc, free zinc ions in the caput, and cauda of epididymis and zinc transporter (ZnT2) decreased significantly, while ZIP12, metallothionein (MT), and metal transcription factor (MTF1) increased significantly. In addition, the expressions of blood-epididymal barrier (BEB)-related molecules (including ZO-1 ß-catenin and N-cadherin) and aquaporins (AQPs, including AQP3, AQP9, and AQP11) in the epididymis of mice in group D were significantly decreased. In addition, compared with groups D, Z, and M, in the ZM group, the expression of BEB-related molecules (including ZO-1, ß-catenin, and N-cadherin) and aquaporins (AQP3, AQP9, and AQP11) in epididymis tissue were significantly increased, and sperm motility and serum testosterone were significantly increased. It was concluded that male diabetic mice have a disturbed epididymal structure and decreased semen quality by causing an imbalance in epididymal zinc homeostasis, BEB, and impaired absorptive function. The combination of zinc and metformin is an effective and safe alternative treatment and provides additional benefits over metformin alone.