Exploring the mystery of colon cancer from the perspective of molecular subtypes and treatment.

The molecular categorization of colon cancer patients remains elusive. Gene set enrichment analysis (GSEA), which investigates the dysregulated genes among tumor and normal samples, has revealed the pivotal role of epithelial-to-mesenchymal transition (EMT) in colon cancer pathogenesis. In this study, we employed multi-clustering method for grouping data, resulting in the identification of two clusters characterized by varying prognostic outcomes. These two subgroups not only displayed disparities in overall survival (OS) but also manifested variations in clinical variables, genetic mutation, and gene expression profiles. Using the nearest template prediction (NTP) method, we were able to replicate the molecular classification effectively within the original dataset and validated it across multiple independent datasets, underscoring its robust repeatability. Furthermore, we constructed two prognostic signatures tailored to each of these subgroups. Our molecular classification, centered on EMT, hold promise in offering fresh insights into the therapy strategies and prognosis assessment for colon cancer.

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.

Combined Analysis of the Transcriptome, Proteome and Metabolome in Human Cryopreserved Sperm.

PURPOSE: This study aimed to identify the altered pathways and genes associated with freezing damage in human sperm during cryopreservation by multiomics analysis. MATERIALS AND METHODS: Fifteen fresh human semen samples were collected for transcriptomic analysis, and another 5 fresh human semen samples were obtained for metabolomic analysis. For each semen sample, 1 mL was cryopreserved, and another 1 mL was left untreated for paired design. The results were then combined with previously published proteomic results to identify key genes/pathways. RESULTS: Cryopreservation significantly reduced sperm motility and mitochondrial structure. Transcriptomic analysis revealed altered mitochondrial function, including changes in tRNA-methyltransferase activity and adenosine tri-phosphate/adenosine di-phosphate transmembrane transporter activity. Metabolomic analysis showed that the citrate cycle in mitochondria was significantly altered. Combining transcriptomic, proteomic, and metabolomic analyses revealed 346 genes that were altered in at least two omics analyses. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that metabolic pathways were significantly altered and strongly associated with mitochondria. Five genes were altered in all three omics analyses: COL11A1, COL18A1, LPCAT3, NME1, and NNT. CONCLUSIONS: Five genes were identified by multiomics analysis in human cryopreserved sperm. These genes might have specific functions in cryopreservation. Explorations of the functions of these genes will be helpful for sperm cryopreservation and sperm motility improvement or even for reproduction in the future.

Novel biallelic variants in DNAH1 cause multiple morphological abnormalities of sperm flagella with favorable outcomes of fertility after ICSI in Han Chinese males.

BACKGROUND: Multiple morphological abnormalities of sperm flagella is an idiopathic asthenoteratozoospermia characterized by absent, short, coiled, angulation, and irregular-caliber flagella. Genetic variants of DNAH1 gene have been identified as a causative factor of multiple morphological abnormalities of sperm flagella and intracytoplasmic sperm injection is an available strategy for infertile males with dynein axonemal heavy chain 1 defects to conceive. OBJECTIVES: To identify novel variants and candidate mutant hotspots of DNAH1 gene related to multiple morphological abnormalities of sperm flagella and male infertility in humans. MATERIALS AND METHODS: The DNAH1 variants were identified by whole exome sequencing and confirmed with Sanger sequencing. Papanicolaou staining, scanning and transmission electron microscopy, and immunostaining were performed to investigate the morphological and ultrastructural characteristics of spermatozoa. Intracytoplasmic sperm injection was applied for the assisted reproductive therapy of males harboring biallelic DNAH1 variants. RESULTS: We identified 18 different DNAH1 variants in 11 unrelated families, including nine missense variants (p.A2564T, p.T3657R, p.G1862R, p.L2296P, p.T4041I, p.L611P, p.A913D, p.R1932Q, p.R2356W) and nine loss-of-function variants (c.2301-1G>T, p.Q1518*, p.R1702*, p.D2845Mfs*2, p.P3909Rfs*33, p.Q4040Dfs*33, p.Q4058*, p.E4060Pfs*61, p.V4071Cfs*54). A total of 66.7% (12/18) of the identified variants were novel. Morphological analysis based on Papanicolaou staining and scanning electron microscopy demonstrated the typical multiple morphological abnormalities of sperm flagella characteristics of dynein axonemal heavy chain 1-deficient spermatozoa. Immunostaining further revealed the absence of inner dynein arms but not outer dynein arms, which induced a general ultrastructural disorganization, such as the loss of central pair and mis-localization of the microtubule doublets and outer dense fibers. To date, seven affected couples have accepted the intracytoplasmic sperm injection treatment, and three of them have given birth to five healthy babies. DISCUSSION AND CONCLUSION: These findings further expand the variant spectrum of DNAH1 gene related to multiple morphological abnormalities of sperm flagella and male infertility in humans, thus providing new information for the molecular diagnosis of asthenoteratozoospermia. The favorable fertility outcomes of intracytoplasmic sperm injection will facilitate the genetic counseling and clinical treatment of infertile males with multiple morphological abnormalities of sperm flagella in the future.

Effects of Qilin pills on spermatogenesis, reproductive hormones, oxidative stress, and the TSSK2 gene in a rat model of oligoasthenospermia.

BACKGROUND: Qilin pills (QLPs), a classic Traditional Chinese Medicine (TCM) formula for treating male infertility, effectively improve semen quality in clinical trials. This study was designed to evaluate the effects of QLPs on spermatogenesis, reproductive hormones, oxidative stress, and the testis-specific serinekinase-2 (TSSK2) gene in a rat model of oligoasthenospermia. METHODS: Forty adult male Sprague-Dawley (SD) rats were randomly divided into four groups. The rat model with oligoasthenospermia was generated by intragastric administration of tripterygium glycosides (TGs) once daily for 4 weeks. Then, two treatment groups were given different doses (1.62 g/kg and 3.24 g/kg) of QLPs once daily for 60 days. Sperm parameters, testicular histology and reproductive hormone measurements, oxidative stress tests, and TSSK2 expression tests were carried out. RESULTS: QLPs effectively improved semen parameters and testicular histology; restored the levels of FSH, LH, PRL, fT, and SHBG; reduced the levels of oxidative stress products (ROS and MDA); increased testicular SOD activity; and restored the expression of spermatogenesis-related gene TSSK2. CONCLUSION: QLPs have a therapeutic effect on a rat model of oligoasthenospermia, and this effect is manifested as improvement of semen quality and testis histology, gonadal axis stability, decreased oxidative stress, and the regulation of testis-specific spermatogenesis-related gene TSSK2.

[Impact of the freezing-thawing process on human sperm mitochondria].

OBJECTIVE: To study the structure and function of human sperm mitochondria before and after the freezing-thawing process. METHODS: Human sperm from healthy donors were subjected to the slow freezing-thawing process, and the sperm mitochondrion-related indexes compared before and after cryopreservation. The ultrastructural changes of the mitochondria were observed under the projection electron microscope, the mitochondrial membrane potential (MMP) and seminal adenosine triphosphate (ATP) content measured by immunofluorescence labeling and ELISA, respectively, and the sperm oxidative stress related indexes detected before and after sperm cryopreservation. RESULTS: Electron microscopy showed loose structures and widened crests of the sperm mitochondria, some with vacuole-like changes after the freezing-thawing process. The sperm after cryopreservation, compared with those before it, exhibited significantly increased contents of oxygen free radicals (ï¼»11.6 ± 3.8ï¼½% vs ï¼»9.6 ± 4.1ï¼½%, P < 0.05) and malondialdehyde (ï¼»3.2 ± 1.4ï¼½ vs ï¼»2.3 ± 1.2ï¼½ nmol/108, P < 0.05), but decreased antioxidant capacity (ï¼»0.6 ± 0.4ï¼½ vs ï¼»0.9 ± 0.4ï¼½ nmol/108, P < 0.05), superoxide dismutase activity (ï¼»0.9 ± 0.4ï¼½ vs ï¼»9.1 ± 3.9ï¼½ nmol/108, P < 0.05), MMP (ï¼»52.2 ± 6.2ï¼½% vs ï¼»55.7 ± 4.9ï¼½%, P = 0.026) and ATP production (ï¼»56.5 ± 9.0ï¼½ vs ï¼»61.3 ± 10.4ï¼½ pmol/106, P = 0.014). CONCLUSIONS: The freezing-thawing process can cause ultrastructural disorder of human sperm mitochondria, reduce their membrane potential and decrease their ATP production.

Sperm Cryopreservation for Male Cancer Patients: More than 10 Years of Experience, in Beijing China.

BACKGROUND Fertility preservation is very important for male cancer patients, especially adolescents. Unfortunately, the use of fertility preservation is very low among Chinese male cancer patients. Additionally, the cumulative rate of frozen sperm use is also low. MATERIAL AND METHODS We performed a retrospective study by collecting available information at the Human Sperm Bank, National Research Institute for Family Planning from July 2006 to December 2017 to examine the data in China. RESULTS A total 145 male cancer patients underwent sperm cryopreservation. The patients were 29.3±6.9 years old, and 6.2% (9 out of 145) of the patients were adolescents under the age of 18 years old. As of June 2018, only 9.7% (14 out of 145) of patients returned to use their cryopreserved sperm for assisted reproduction technology (ART). Of the 33 ART cycles, conceptions were achieved in 51.5% (17 out of 33), and the rate of patients who had a baby was 71.4% (10 out of 14). The data indicate men with testicular cancer or leukemia had lower total sperm counts and recovery rate of progressive sperm than did men with other types of cancer, while men with sarcoma had the lowest progressive sperm. CONCLUSIONS The physician should make an effort to promote fertility preservation for male cancer patients in China. And patients with testicular cancer and leukemia require additional attention.

[Status quo of autologous sperm preservation in the human sperm bank in Beijing].

OBJECTIVE: To comprehensively analyze the status quo of autologous sperm preservation in the human sperm bank in Beijing and better utilize the existing resources for the preservation of male fertility. METHODS: We retrospectively analyzed the geographical data and semen quality of 251 males with autologous sperm preservation in the Human Sperm Bank, Science and Technology Research Institute, National Health and Family Planning Commission of China from July 2006 to December 2016. RESULTS: The rate of autologous sperm preservation in the Human Sperm Bank was as low as 8.76% between July 2006 and December 2010 but increased annually by 119% on average from 2011 to 2013. Of the 251 males involved, 204 (81.27%) were aged 20－39 years, 175 (69.72%) had bachelor's or master's degree, 223 (88.84%) had no child, 69 (27.49%) got less than 10 tubes of semen samples frozen, and 26 (10.36%) had their semen samples cryopreserved only once. The utilization rate of the cryopreserved sperm was only 5.58 % (n = 14). The main reason for autologous sperm preservation was carcinoma (55.78% ï¼»n = 140ï¼½), including blood cancer (22.31% ï¼»n = 66ï¼½), testicular cancer (13.15% ï¼»n = 33ï¼½) and other cancers (16.33% ï¼»n = 41ï¼½). Compared with the non-cancer males, the cancer patients had a significantly reduced mean sperm concentration (90.45 vs 60.53 ×106/ml, P < 0.05), total sperm count (311.3 vs 175.8 ×106, P < 0.05), percentage of progressively motile sperm (PMS) (49.21% vs 43.55%, P < 0.05) and recovery rate of PMS (68.13% vs 52.17%, P < 0.05). In the subgroups of testicular, blood and other cancers, the sperm concentration averaged 37.68, 57.98 and 90.69 ×106/ml, the semen volume 2.73, 2.82 and 3.41 ml, the total sperm count 93.29, 158.41 and 349.49 ×106, the percentage of PMS 45.32%, 43.47% and 44.49%, and the recovery rate of PMS 48.32%, 50.07% and 61.09%, respectively, the sperm concentration and total sperm count significantly lower in the testicular cancer patients than in the other two groups (P < 0.05). CONCLUSIONS: The number of the cases of autologous sperm preservation in Beijing is increasing year by year, and the majority of them are cancer patients. As most of the cancer patients have missed the best period for sperm preservation, sperm bank workers should endeavor to increase the public awareness of autologous sperm preservation.

Qilin pills alleviate oligoasthenospermia by inhibiting Bax-caspase-9 apoptosis pathway in the testes of model rats.

At present, the treatment of oligoasthenospermia with western medicine is ineffective. Qilin pill (QLP) is a Chinese traditional medicine for treating male infertility. Recent multicenter clinical studies in China reported that QLPs markedly improved sperm quality. However, the mechanism of action of QLPs on oligoasthenospermia remains unknown. In this study, we investigated the mechanistic basis for improvement of semen parameters and reversal of testis damage by QLPs in a rat model of oligoasthenospermia induced by treatment with tripterygium glycosides (TGs) (40 mg/kg) once daily for 4 weeks. Rats were administered QLPs (1.62 g/kg or 3.24 g/kg) each day for 60 days, with untreated animals serving as controls. The concentration and motility of sperm extracted from rat epididymis were determined, whereas histopathological examination and immunohistochemical apoptosis analysis of rat testes was performed. Expression profiles of apoptosis-related genes were determined by microarray analysis; the results were validated by quantitative real-time PCR, western blotting, and immunohistochemistry. Sperm concentration and motility in the QLP treatment group were increased relative to those in control rats. Testis tissue and DNA damage were reversed by QLP treatment. The improvement function of QLPs on sperm and testis works mainly by suppressing mitochondrial apoptosis in the testis via modulation of B cell lymphoma (Bcl)-2, Bcl-2-associated X protein (Bax), cytochrome C, caspase-9 and caspase-3 expression. QLPs could improve sperm quality and testis damage in a rat model of oligoasthenospermia by inhibiting the Bax-Caspase-9 apoptosis pathway and exerting therapeutic effects.

[Levels of serum reproductive hormones and metabolic syndrome in adult males].

OBJECTIVE: To investigate the prevalence of metabolic syndrome (MS) and its relationship with reproductive hormones in adult males in China. METHODS: Using the cluster and stratified sampling methods, we conducted an investigation among 3 600 adult males aged over 20 years in Hebei, Shaanxi and Jiangsu provinces. We obtained their reproductive hormone levels, biochemical indicators and basic body indexes and compared them between the two groups of subjects. RESULTS: A total of 3 332 valid serum samples were collected, which revealed a prevalence rate of MS of 38.5% in the adult males. Compared with the non-MS subjects, the MS males showed a significantly higher free testosterone index (FTI, 0.39 ± 0.15 vs 0.45 ± 0.19, P < 0.01) but lower levels of total testosterone (TT, ï¼»16.35 ± 4.78ï¼½ vs ï¼»13.37 ± 4.23ï¼½ nmo/L, P < 0.01) and sex hormone-binding globulin (SHBG, ï¼»47.13 ± 20.50ï¼½ vs ï¼»33.32 ± 14.91ï¼½ nmol/L, P < 0.01) and testosterone secretion index (TSI, 3.64 ± 1.92 vs 3.14 ± 1.80, P < 0.01). There were no statistically significant differences between the two groups in the levels of calculated free testosterone (cFT) and LH (P > 0.05). Spearman rank correlation analysis showed that the levels of serum TT and SHBG were correlated with all the indicators of MS (P < 0.01) except systolic blood pressure (SBP), while that of cFT only with the levels of high-density lipoprotein (HDL), fasting blood glucose (FBG) and SBP (P < 0.01). After adjusted for age, smoking, drinking, and body mass index (BMI), all the MS indicators were significantly associated with the SHBG level (P < 0.01), but not high blood pressure with the serum TT level (P > 0.05). After adjusted for the age, smoking, drinking, BMI and TT, the serum SHBG level was the main independent risk factor for MS (OR: 0.965, 95% CI: 0.958－0.973, P < 0.01). CONCLUSIONS: The level of serum cFT is not correlated with while that of SHBG is the main independent risk factor for metabolic syndrome in adult males in China.

Tris (2-chloroethyl) phosphate induces senescence-like phenotype of hepatocytes via the p21Waf1/Cip1-Rb pathway in a p53-independent manner.

Tris (2-chloroethyl) phosphate (TCEP) has been widely used as a plasticizer and flame retardant. TCEP as a potential carcinogen is often detected in the occupational and nature environments. To investigate effects of TCEP on human hepatocytes, we assessed cell growth rate, cellular membrane integrity, senescence-associated ß-galactosidase (SA-ß-Gal) activity and analyzed expression of regulators involved in the p53-p21Waf1/Cip1-Rb pathway in TCEP-treated L02 cells. The results showed TCEP increased the percentage of SA-ß-Gal positive cells, decreased IL-6 levels, down-regulated the regulators of p38MAPK-NF-κB pathways, but up-regulated the regulators of p21Waf1/Cip1-Rb pathway in L02 cells. Furthermore, we measured the SA-ß-Gal activity and expression of regulators involved in the p53-p21Waf1/Cip1-Rb pathway in L02-p53 cells and p53-null Hep3B cells. Similar results were found in L02-p53 cells and Hep3B cells. The findings demonstrated that TCEP induced senescence-like growth arrest via the p21Waf1/Cip1-Rb pathway in a p53-independent manner, without activation of the IL-6/IL6R, p38MAPK-NF-κB pathways in hepatocytes.

Tris(2-chloroethyl)phosphate-induced cell growth arrest via attenuation of SIRT1-independent PI3K/Akt/mTOR pathway.

Tris(2-chloroethyl)phosphate (TCEP) as an organophosphorus flame retardant and plasticizer has been widely used in industrial and household products. It not only was detected in residential indoor air and dust, surface and drinking water, but also in human plasma and breast milk, and tissue samples of liver, kidneys and brain from rodents. TCEP is classified as carcinogenic category 2 and toxic for reproduction category 1B. Sufficient evidence from experimental animals indicated carcinogenicity of TCEP in the liver, and kidneys as well as cell loss in the brain. However, the underlying mechanisms of TCEP-induced hepatotoxicity are mostly unknown. We investigated the in vitro effects of TCEP as well as TCEP-induced cell growth in the L02 and HepG2 cells through the PI3K/Akt/mTOR pathway. We found that TCEP reduced cell viability of these cell lines, induced the cell growth arrest, upregulated mRNA and protein levels of SIRT1, and attenuated the PI3K/Akt/mTOR pathway. However, growth arrest of the L02 and HepG2 cells were aggravated after inhibiting the SIRT1 expression with EX-527. The findings above suggested that TCEP induced the cell growth arrest of L02 and HepG2 cells via attenuation of the SIRT1-independent PI3K/Akt/mTOR pathway. Copyright © 2015 John Wiley & Sons, Ltd.

Is serum sex hormone-binding globulin a dominant risk factor for metabolic syndrome?

This multi-center, cross-sectional study investigated the association between serum testosterone (T) levels, serum sex hormone-binding globulin (SHBG) levels, and the risk of metabolic syndrome (MS) in 3332 adult Chinese men. The prevalence of MS was 34.7%, and men with MS had lower serum levels of total T (TT) and SHBG than those without MS (P < 0.001). There was no significant difference in serum free T (FT) levels between subjects with and without MS (P = 0.627). In logistic regression analysis, the association between MS and serum SHBG levels persisted after adjusting for age, body mass index (BMI), smoking and drinking status, and serum TT (odds ratio [OR] 0.962, 95% confidence interval [95% CI] 0.954-0.969, P< 0.01). However, the association between serum TT level and the risk of MS was weak after adjusting for age, BMI, SHBG level, and smoking and drinking status (OR 0.981, 95% CI 0.960-1.007). Our study reveals that both serum TT and SHBG levels, but not serum FT, are inversely associated with the prevalence of MS and that serum SHBG is an independent and dominant risk factor for MS.

[Effect of peer education on postoperative rehabilitation of patients with laryngeal cancer].

OBJECTIVE: To evaluate the effect of peer education on postoperative rehabilitation of patients with laryngeal cancer. METHODS: From January 2010 to December 2012, 240 cases of primary laryngeal cancer were treated with partial or total laryngectomy as well as tracheotomy in the Department of Head and Neck Surgery in Shandong Provincial Hospital. The patients were randomly divided into experimental group or control group by Stochastic tables law, 120 patients in control group accepted only conventional nursing, 120 patients in experimental group accepted both conventional nursing and peer education. The difference in the treatment nursing cooperation attitude, postoperative recovery, postoperative symptoms and the incidence of complications between two groups were evaluated by postoperative quality of life (UW-QOL), Eysenck personality questionnaire (EPQ) and coping style questionnaire (CSQ) score. The independent sample T-test and χ² test were used for statistical analysis. RESULTS: The patients of experimental group showed more positive attitude to treatment and care compared to the patients of control group (χ² = 10.7, P < 0.01). Experimental group had less time in postoperative wound healing time, gastric tube indwelling time and postoperative hospital stay than control group, with statistically significant differences (t = 6.89, t = 6.36, t = 6.42, respectively, P < 0.01); Incidence of postoperative discomfort symptoms in experimental group was less than that in control group (P < 0.05). Total score for quality of life (UW-QOL) in experimental group was higher than that in control group (t = 2.40, P < 0.05). The P and N scores of EPQ in experimental group were lower than those in control group (t = 2.28, t = 2.60, P < 0.05), while the E score of EPQ in experimental group was significantly higher than that in control group (t = 4.50, P < 0.01), with no significant difference in the L score of EPQ between two groups (P > 0.05). scores of factor to solve problem and help factor of CSQ in experimental group were higher than those in control group (t = 7.05, t = 7.73, P < 0.01), and the era factor score of experimental group was lower than that of control group (t = 2.05, P < 0.05). CONCLUSIONS: Peer education can help the patients with laryngectomy to maintain good psychological and mental status, to cooperate actively with treatment, to reduce symptoms and complications, and to shorten the length of hospital stay, improving their postoperative rehabilitation and quality of life.

MEHP-induced oxidative DNA damage and apoptosis in HepG2 cells correlates with p53-mediated mitochondria-dependent signaling pathway.

In the present study, the effects of MEHP on human hepatocellular liver carcinoma HepG2 cells were investigated. The results showed that MEHP-induced oxidative DNA damage in the treatment groups (≥ 25.00 µM) at 24h after treatment and in the 100.00 µM treatment group at 36 h after treatment (p < 0.05 or p < 0.01). At 36 h after treatment, MEHP at higher concentrations (≥ 25.00 µM) resulted in a decrease in ATP level, and an increase in the protein levels of cytochrome c and Smac/DIABLO in the cytosol as well as the percentage of apoptotic cells. The activation of caspase-9 and -3 and the expression of the selected apoptosis-related proteins, p53, PUMA, NOXA, Bax and Bcl-2 were also induced. Furthermore, vitamin C, a scavenger of reactive oxygen species, attenuated MEHP-induced apoptosis. These findings indicated that MEHP induced oxidative DNA damage and apoptosis in HepG2 cells, and p53 and its downstream proteins were involved in mitochondria- and caspase-mediated apoptosis induced by MEHP.

[Clinical validation of screening scales for late onset of hypogonadism in Chinese males].

OBJECTIVE: To investigate the incidence of late onset of hypogonadism (LOH) in males as well as the sensitivity, specificity and applicability of the androgen deficiency in aging males (ADAM) questionnaire and the aging males' symptoms (AMS) scale in Chinese males based on the community population data. METHODS: A total of 1 498 old and middle-aged healthy males attended the ADAM and AMS investigations, of whom 434 received the measurement of the concentration of serum reproductive hormones, the positive rate of LOH screening, the rate of androgen deficiency, the clinical incidence of LOH, and the sensitivity and specificity of ADAM and AMS by tT and fT cut-off. The symptom evaluation cut-off value of the AMS score was optimized using the ROC curve. RESULTS: Although 5 of the subjects had previously received irregular testosterone supplement, none of them was given or actively sought for androgen therapy at the time of the investigation. Among the 40 - 69 years old males, the mean positive rates of LOH screening by ADAM and AMS were 80.77% and 32.34%, and the mean androgen deficiency rates obtained by tT and fT cut-off were 14.02% and 43.69%. The mean clinical incidences of LOH in the ADMA- and AMS-positive subjects were 37.85% and 15.42%. According to the fT cut-off, the sensitivities of ADAM and AMS were 86.63% and 35.29%, and their specificities were 24.48% and 63.49%, respectively. The symptom evaluation cut-off value of the AMS score optimized by the ROC curve was 19.5. CONCLUSION: The sensitivity and specificity of ADAM and AMS in the Chinese population are basically consistent with the results of most studies abroad, while the positive rate of LOH screening, the rate of androgen deficiency and the clinical incidence of LOH obviously higher in the former than those reported in other studies. Both ADAM and AMS are applicable to the Chinese population. The former is advantageous for its high sensitivity, time saving and easy operation, and therefore suitable for screening LOH, while the latter can be used for monitoring therapeutic efficacy.

Elevated levels of urinary 8-hydroxy-2 -deoxyguanosine, lymphocytic micronuclei, and serum glutathione S-transferase in workers exposed to coke oven emissions.

To investigate associations among occupational exposure to coke oven emissions (COEs), oxidative stress, cytogenotoxic effects, change in the metabolizing enzyme glutathione S-transferase (GST), and internal levels of polycyclic aromatic hydrocarbons (PAHs) in coke oven workers, we recruited 47 male coke oven workers and 31 male control subjects from a coke oven plant in northern China. We measured the levels of 1-hydroxypyrene (1-OHP) and 8-hydroxy-2 -deoxyguanosine (8-OHdG) in urine, micronucleated binucleated cells (BNMNs) in peripheral blood lymphocyte, and GST in serum. Our results showed that the group exposed to COEs had significantly increased levels of 1-OHP [median 5.7; interquartile range (IQR), 1.4-12.0 micromol/mol creatinine] compared with the control group (3; 0.5-6.4 micromol/mol creatinine). In addition, the median levels (IQR) of 8-OHdG, BNMNs, and GST were markedly increased in the exposed [1.9 (1.4-15.4) micromol/mol creatinine; 6 (2-8) per thousand ; 22.1 (14.9-31.2) U/L, respectively] compared with controls [1.3 (1.0-4.0) micromol/mol creatinine, 2 (0-4) per thousand; and 13.1 (9.5-16.7) U/L, respectively]. These results appeared to be modified by smoking. However, multivariate logistic regression analysis revealed that exposure to COEs had the highest odds ratio among variables analyzed and that smoking was not a significant confounder of the levels of studied biomarkers. Overall, the present findings suggest that COE exposure led to increased internal PAH burden, genetic damage, oxidative stress, and GST activity. The consequences of the changes in these biomarkers, such as risk of cancer, warrant further investigations.

[Analysis of serum glutathione S-transferase and urinary 8-hydroxy-2-deoxyguanosine in coke oven workers].

OBJECTIVE: To investigate the application of serum glutathione S-transferase (GST) and urinary 8-hydroxy-2-deoxyguanosine (8-OHdG) as the monitoring biomarkers for coke oven workers exposed to polycyclic aromatic hydrocarbons (PAHs). METHODS: 47 male coke oven workers and 31 male control workers were investigated. Urinary 8-OHdG and serum GST were analyzed using high performance liquid chromatography (HPLC) with electrochemical detection and test kit. Urinary 1-hydroxypyrene (1-OHP) as internal exposure of PAHs was also determined simultaneously by alkaline hydrolysis and HPLC. RESULTS: The values of urinary 1-OHP, serum GST and urinary 8-OHdG were reported as median with interquartile range (P(25)-P(75)). Urinary 1-OHP [5.7 (1.4-12.0) micromol/mol Cr], serum GST [22.1 (14.9-31.2) U/ml], and urinary 8-OHdG [1.9 (1.4-15.4) micromol/mol Cr] in coke oven workers were significantly higher than in control workers [3.0 (0.5-6.4) micromol/mol Cr (P < 0.05), 13.1 (9.5-16.7) U/ml (P < 0.01), and 1.3 (1.0-4.0) micromol/mol Cr (P < 0.05) respectively]. Categorizing by smoking status, significant differences in urinary 1-OHP and serum GST were found only in smokers among coke oven workers compared to control workers (P < 0.01), and 8-OHdG levels only in non-smokers (P < 0.01). Additionally, there was significant correlation between urinary 1-OHP and serum GST activity (r(s) = 0.31, P < 0.01, n = 78). The multiple logistic regression analysis showed that coke oven workers were at the higher risk of having GST activities above 16.7 U/ml (OR = 13.2) and 8-OHdG levels above 1.8 micromol/mol creatinine (OR = 4.4). High body mass index was an independent factor to affect urinary 8-OHdG levels. CONCLUSIONS: The elevated serum GST activities and increased oxidative DNA damage were found in the coke oven workers. Occupational exposure and smoking interact on each other. Serum GST may be used as a biomarker for assessing the exposure of PAHs. Assay of urinary 8-OHdG may be useful for evaluating the risk of lung cancer in coke oven workers.

DNA repair gene XRCC1 polymorphisms, smoking, and esophageal cancer risk.

To investigate the effect of X-ray repair cross complementing 1 (XRCC1) genetic polymorphisms on esophageal cancer risk, we determined XRCC1 polymorphisms at codon 194 (Arg --> Trp) and codon 399 (Arg --> Gln) in 135 patients with esophageal squamous cell carcinoma (ESCC) and 152 normal controls from hospitals. Although polymorphism at codon 194 was not associated with risk for ESCC, we found that the frequency of XRCC1 399 Gln/Gln genotype in ESCC patients (14.1%) was significantly higher than that in normal controls (3.3%), and that XRCC1 399 Gln/Gln genotype was associated with an increased risk of ESCC (odds ratio (OR) = 5.15, 95% confidence interval (CI): 2.42-0.93). In addition, we found that the risk for smoker increased 4.2-fold than non-smokers in the 399 Gln/Gln genotype (OR = 4.20, 95% CI: 2.37-7.44). These results suggest that XRCC1 399 Gln/Gln genotype may contribute to the risk of ESCC and modify risk associated with smoking.

Expression of cyclooxygenase-2 (COX-2) in human esophageal cancer and in vitro inhibition by a specific COX-2 inhibitor, NS-398.

BACKGROUND: The purpose of the present paper was to study the expression of cyclooxygenase-2 (COX-2) in normal squamous epithelium, squamous dysplasia and squamous cell carcinoma (SCC) of the esophagus, to elucidate the role of COX-2 in esophageal carcinogenesis, and to evaluate the in vitro effect and mechanism of a COX-2 inhibitor, NS-398, in inducing growth inhibition and apoptosis of human esophageal cancer cells. METHODS: Biopsy specimens of esophageal dysplasia (n = 21), and surgical resections of SCC (n = 37) were compared with normal esophagus (n = 37) and analyzed by RT-PCR. Human esophageal cells were used for the study. Anti-proliferative effect was measured by MTT, apoptosis was determined by DNA fragmentation assay. RESULTS: Marked COX-2 expression was shown in SCC and esophageal squamous dysplasia, and no marked COX-2 expression was observed in the normal squamous epithelium, respectively. NS-398 could inhibit esophageal cells growth in a dose-dependent manner, induce apoptosis, and elevate caspase-3 activity in vitro. CONCLUSIONS: This study provides evidence that COX-2 is upregulated in the majority of cases of squamous dysplasia and SCC of esophagus, and that NS-398 can inhibit growth and induce apoptosis via activating caspase-3 activity in vitro. These results suggest that selective inhibitors of COX-2 may be an effective preventive and therapeutic option for esophageal carcinoma.