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Background information Diabetes-induced testicular dysfunction is characterised by abnormal apoptosis of spermatogenic cells, but the underlying mechanism is poorly understood. This study aimed to investigate the roles of clusterin (CLU) in testicular damage associated with diabetes pathogenesis, as well as the molecular mechanism. A rat diabetes model was established using streptozocin, and the mouse spermatogenic cell line GC-1 spg was treated with high glucose as a cellular model. CLU was overexpressed in GC-1 spg cells, followed by detection of serum testosterone, cell proliferation, cell apoptosis and autophagy. Results CLU expression was significantly reduced and LC3 expression was elevated in testis tissues in the rat diabetes model and high glucose-treated GC-1 spg cells. High glucose led to suppressed viability, enhanced apoptosis, reduced Bcl-2 expression, elevated Bax expression and cleavage of Caspase-3/-9 in GC-1 spg cells, and these effects were abrogated by CLU overexpression. Additionally, CLU overexpression repressed LC3 and Beclin-1 expression, reduced the LC3II/LC3I ratio and promoted p62 expression in GC-1 spg cells in the presence of high glucose, and these effects were all mitigated by rapamycin treatment. Inhibition of PI3K/AKT/mTOR signalling with LY294002 activated autophagy in CLU-overexpressing GC-1 spg cells under high glucose conditions. CLU overexpression repressed autophagy and alleviated testicular damage in diabetic rats, which was also abrogated by LY294002 treatment. Conclusions CLU expression is suppressed during diabetes-induced testicular damage, whereas CLU overexpression alleviates diabetes-induced testicular damage by activating PI3K/AKT/mTOR signalling to inhibit autophagy and further repress spermatogenic cell apoptosis.

To study the effect of taurine on the reproductive toxicity damage induced by formaldehyde (FM) in adult male rats.Forty-eight SD adult male rats were equally randomized into a normal control, an FM poisoning (FMP), a taurine intervention (TI), and an TI+FMP group. The control rats were given normal diet and gavage of saline, the rats of the FMP group treated intraperitoneally with FM at 10 mg/kg qd alt, those of the TI group intragastrically with taurine at 100 mg/kg qd, and those of the TI+FMP group with both FM and taurine at the above doses. After 30 days of treatment, the blood of the abdominal cardinal vein of the rats was extracted for measurement of the levels of serum hormones, the body weight, testis weight and testicular coefficient obtained, the testis tissue subjected to HE staining, and the expressions of Bcl-2 and Bax determined by Western blot.There were no statistically significant differences among the four groups of rats in the body weight, testis weight or testicular coefficient (P0.05). The rats in the FMP group showed obviously decreased testicular spermatogenic cells and disordered layers and loose structure of seminiferous tubules, which were basically restored to normal after taurine intervention. Compared with the normal controls, the animals of the TI group exhibited no significant abnormality, but those of the FMP group presented markedly reduced levels of serum T, LH and FSH (P0.05), and dramatically increased level of Gonadotropin-releasing hormone (GnRH) (P0.01). The levels of serum hormones were all significantly improved (P0.05) and that of the apoptotic protein Bax basically returned to normal (P0.05) after taurine intervention.Taurine has a certain protective effect against male reproductive toxicity caused by formaldehyde.

Polybrominated biphenyl ether (PBDE) is a kind of brominated aromatic hydrocarbon commonly used as a flame retardant and now considered as one of the persistent organic pollutants, which generally exists in the environment and organisms. Studies have shown that PBDEs have an endocrine disrupting effect, neurotoxicity, developmental toxicity, reproductive toxicity, and the ability of inducing reproductive toxicity in men by reducing semen quality, affecting testis germ cells and interfering with reproductive hormone levels. Although PBDEs have been banned in some countries, they do exist in our environment. Therefore, more studies are needed to reveal their potential toxicity and mechanisms, so as to monitor, predict and prevent their pollution. This review focuses on the male reproductive toxicity of PBDEs.

The enhanced motility of cancer cells via the remodeling of the actin cytoskeleton is crucial in the process of cancer cell invasion and metastasis. It was previously demonstrated that gelsolin (GSN) may be involved as a tumor or a metastasis suppressor, depending on the cell lines and model systems used. In the present study, the effect of GSN on the growth and invasion of human colon carcinoma (CC) cells was investigated using reverse transcription quantitative polymerase chain reaction and western blotting. It was observed that upregulation of the expression of GSN in human CC cells significantly reduced the invasiveness of these cells. The expression levels of GSN were observed to be reduced in CC cells, and the reduced expression level of GSN was often associated with a poorer metastasis‑free survival rate in patients with CC (P=0.04). In addition, the overexpression of GSN inhibited the invasion of CC cells in vitro. Furthermore, GSN was observed to inhibit signal transducer and activator of transcription (STAT) 3 signaling in CC cells. Together, these results suggested that GSN is critical in regulating cytoskeletal events and inhibits the invasive and/or metastatic potential of CC cells. The results obtained in the present study may improve understanding of the functional and mechanistic links between GSN as a possible tumor suppressor and the STAT3 signaling pathway, with respect to the aggressive nature of CC. In addition, the present study demonstrated the importance of GSN in regulating the invasion and metastasis of CC cells at the molecular level, suggesting that GSN may be a potential predictor of prognosis and treatment success in CC.

Supplemental Digital Content is available in the text, Backgrounds and Objective: Several clinical trials have shown that grape seed extract can reduce blood pressure, but the results are often irreproducible. We therefore sought to systematically evaluate the impact of grape seed extract treatment on the changes of systolic/diastolic blood pressure (SBP/DBP) by meta-analyzing available randomized controlled trials. Methods: Trial selection and data extraction were completed independently by 2 investigators. Effect-size estimates were expressed as weighted mean difference (WMD) and 95% confidence interval (CI). Results: Twelve articles involving 16 clinical trials and 810 study subjects were analyzed. Overall analyses found significant reductions for SBP (WMD = −6.077; 95% CI: −10.736 to −1.419; P = 0.011) and DBP (WMD = −2.803; 95% CI: −4.417 to −1.189; P = 0.001) after grape seed extract treatment. In subgroup analyses, there were significant reductions in younger subjects (mean age < 50 years) for SBP (WMD = −6.049; 95% CI: −10.223 to −1.875; P = 0.005) and DBP (WMD = −3.116; 95% CI: −4.773 to −1.459; P

Amiodarone is recognized as an effective drug in the treatment of arrhythmias. Previous experiments demonstrated that mono-N-desethylamiodarone (MDEA) was the major circulating metabolite in humans. In addition, dealkylation, hydroxylation, and deamination were minor metabolic pathways. The purpose of this study was to identify the metabolites of amiodarone in the bile obtained from patients with T-tube drainage after oral drug administration. Amiodarone metabolism in vitro was also investigated using human liver microsomes (HLMs) and S9 fraction. Ultraperformance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF MS) revealed 33 metabolites in human bile, including 22 phase I and 11 phase II metabolites. The major metabolites were MDEA (M7) and ω-carboxylate amiodarone (M12). Metabolite M12 was isolated from human bile, and the chemical structure was confirmed using UPLC-Q/TOF MS and ¹H NMR. Moreover, the authentic standards of two hydroxylated metabolites, 2-hydroxylamiodarone and 3'-hydroxylamiodarone, were obtained through microbial transformation. Several novel metabolic pathways of amiodarone in human were proposed, including ω-carboxylation, deiodination, and glucuronidation. The in vitro study demonstrated that incubation of HLMs with amiodarone did not give rise to any carboxyl metabolites. In contrast, M12 and its metabolites were detected in human liver S9 incubation samples, and the production of these metabolites were inhibited almost completely by 4-methylpyrazole, an inhibitor of alcohol dehydrogenase, suggesting the involvement of alcohol dehydrogenase in the ω-carboxylation of amiodarone. Overall, UPLC-Q/TOF MS analysis leads to the discovery of several novel amiodarone metabolites in human bile and underscores the importance of bile as an excretion pathway.

The aim of the present study was to investigate the effect of anti-estrogen treatment (fulvestrant) on the biological activity of hepatocellular carcinoma (HCC), involving the estrogen receptor α (ERα) and Wnt pathways, and to evaluate whether ERα and Wnt inhibitory factor-1 (WIF1) could be biomarkers for anti-estrogen clinical therapy. H22 and HepG2 cells were treated with 0.04 to 625 nM fulvestrant and the WST-8 method was used to assess the inhibition rate after 72 h. Furthermore, prolactin (PRL) secretion by HepG2 cells was assessed at 24 h using an enzyme immunoassay. Quantitative polymerase chain reaction and western blot analysis were used to analyze the mRNA and protein expression levels of ERα, β-catenin and WIF1, respectively, in HepG2 cells. For clinical patient analysis, the tumor volume was analyzed by magnetic resonance imaging methods, and PRL in the blood was detected by an enzyme immunoassay. In HepG2 cells, the mRNA and protein expression levels of ERα were downregulated (P

Paravertebral block (PVB) has been shown to be an ideal aid for analgesia after thoracic or abdominal surgery. The authors studied the safety and efficacy of the single-dose and bilateral ultrasound-guided (USG)-PVB before combined thoracoscopic-laparoscopic esophagectomy (TLE) along with intravenous sufentanil analgesia as a method of pain relief in comparison with intravenous sufentanil as a sole analgesic agent.Prospective, randomized study.Single university hospital.Fifty-two patients undergoing TLE.A USG-PVB was performed before surgery using a solution of 30 mL of 0.5% ropivacaine by 3 injections of 10 mL each at the right T5 and bilateral T8 (PVB group, n=26) or the saline injection of 10 mL at every site (control group, n=26).Successful PVBs were achieved in all patients of the PVB group. Intraoperative mean remifentanil usage and end-tidal sevoflurane concentration were lower in the PVB group (p0.001). Hemodynamic parameters were stable in both groups. Postoperative pain scores both at rest and on coughing were lower during the first 8 hours in the PVB group than those in the control group (p0.05). Cumulative sufentanil consumption delivered by patient-controlled analgesia (PCA) was significantly lower in the PVB group at all time points (p0.05). Postoperative pulmonary function was better at the third postoperative day in the PVB group (p0.05), with quicker hospital discharge and lower hospital costs (p0.05).The single-dose and bilateral PVB given before TLE combined with sufentanil may provide better postoperative analgesia and early discharge in patients undergoing TLE.

To investigate the role of endothelial progenitor cells (EPC) in the injury of rat optic nerve.An experimental study. The rat model of optic nerve injury was created by fluid percussion brain injury device (FPI). On hundred and eight rats (108 eyes) were divided into 2 groups randomly. Each group was further divided into 9 subgroups by the time of injury (24 h before and 3, 12, 24, 48, 72 h, 1, 2 and 3 weeks after the injury). The number of circulating EPCs was measured, HE staining of the optic nerve, immunohistochemistry study of CD31 (markers of vascular endothelial cells) and flash-visual evoked potential (F-VEP) were observed at every time point. Two independent sample t-test was used for the comparison between the control group and the optic nerve injury groups at the same time point. The correlation between different items was analyzed by Pearson test. P value less than 0.05 was considered significant.The number of EPCs in normal rats was 46-52/200 000 monocytes. After traumatic optic nerve injury, the number of EPCs was (34 ± 4, 34 ± 5, 69 ± 9, 76 ± 6, 107 ± 9, 69 ± 7, 58 ± 6 and 56 ± 4)/200 000 monocytes at 3, 12, 24, 48, 72 h, and 1, 2 and 3 weeks. The difference of number of EPCs between the experiment and control groups was significant at 3, 12, 24, 48, 72 h and 1 and 2 weeks after the injury (t = 5.29, 2.90, -4.30, -7.61, -14.17, -5.74 and -2.79; P0.05). The number of CD31(+) cell in the optic nerve and surround tissues in normal rats was (7-9)/5 high magnification field. After the injury, the number of CD31(+) cell was 8.36 ± 1.52, 7.17 ± 1.10, 10.41 ± 1.92, 11.43 ± 1.58, 14.29 ± 2.03, 17.33 ± 1.47, 17.86 ± 1.22 and 18.13 ± 1.40 at different time points. The difference of number of CD31(+) cell between the experiment and control groups was significant at 48, 72 h, and 1, 2, and 3 weeks after the injury (t = 4.31, -7.61, -8.17, -10.08, and -10.79; P0.05). The number of microvessels in the optic nerve and surround tissues in normal rats was 6-9/5 high magnification field. After traumatic optic nerve injury, the number of microvessels was 7.54 ± 2.01, 8.52 ± 2.21, 11.02 ± 1.62, 15.40 ± 2.04, 18.39 ± 1.96, 23.21 ± 1.50, 22.78 ± 2.40 and 24.13 ± 2.51 at different time points. The difference of number of microvessels between the experiment and control groups was significant at 48, 72 h, 1, 2, and 3 weeks after the injury (t = 4.25, -7.74, -8.26, -10.28 and -11.49; P0.05). The latency period of P waves was decreased at 3 h and increased to above basic level at 24 h, and then tend to be stable. The difference of latency period of P waves between the experiment and control groups was significant at 3, 12, 24, 48, 72 h, 1, 2 and 3 weeks after the injury (t = 4.15, 3.74, 5.84, 6.08, 6.40, 6.52, 6.53 and 6.61; P0.05). The amplitude of F-VEP was decreased at 3 h and increased to the basic level at 12 h, then decreased to below the basic level gradually. The difference of the amplitude of F-VEP between the experiment and control groups was significant at 3, 24, 48, 72 h, 1, 2 and 3 weeks after the injury (t = 3.95, 4.14, 5.26, 5.78, 6.49, 6.72 and 6.23; P0.05). The number of EPCs was correlated with the number of CD31(+) cell, microvessels and F-VEP (r = 0.43, 0.41 and 0.43; P0.01).The present study showed that the number of EPCs in the blood increases significantly after traumatic optic nerve injury, and the cells can arrive the traumatic area to repair injured tissue and enhance angiogenesis.

April 1, 2011, The 1000 Genomes Project aims to provide a deep characterization of human genome sequence variation as a foundation for investigating the relationship between genotype and phenotype. Here we present results of the pilot phase of the project, designed to develop and compare different strategies for genome-wide sequencing with high-throughput platforms. We undertook three projects: low-coverage whole-genome sequencing of 179 individuals from four populations; high-coverage sequencing of two mother–father–child trios; and exon-targeted sequencing of 697 individuals from seven populations. We describe the location, allele frequency and local haplotype structure of approximately 15 million single nucleotide polymorphisms, 1 million short insertions and deletions, and 20,000 structural variants, most of which were previously undescribed. We show that, because we have catalogued the vast majority of common variation, over 95% of the currently accessible variants found in any individual are present in this data set. On average, each person is found to carry approximately 250 to 300 loss-of-function variants in annotated genes and 50 to 100 variants previously implicated in inherited disorders. We demonstrate how these results can be used to inform association and functional studies. From the two trios, we directly estimate the rate of de novo germline base substitution mutations to be approximately 10[superscript −8] per base pair per generation. We explore the data with regard to signatures of natural selection, and identify a marked reduction of genetic variation in the neighbourhood of genes, due to selection at linked sites. These methods and public data will support the next phase of human genetic research., Ministry of Science and Technology of the People's Republic of China (973 program no. 2011CB809200), National Natural Science Foundation (China) (30725008), National Natural Science Foundation (China) (30890032), National Natural Science Foundation (China) (30811130531), National Natural Science Foundation (China) (30221004), China (Chinese 863 program 2006AA02Z177), China (Chinese 863 program 2006AA02Z334), China (Chinese 863 program 2006AA02A302), China (Chinese 863 program 2009AA022707), National Institutes of Health (U.S.) (Grant U54HG2750), National Institutes of Health (U.S.) (Grant U01HG5208), National Institutes of Health (U.S.) (Grant U54HG3067)

To explore the morbidity, burdens caused and risk factors of injuries among children and to provide information for policy making on prevention.By PPS sampling method, we selected 28 000 households in Beijing and using a questionnaire developed by UNICEF, TASC and China CDC.The morbidity of child injury in Beijing was 2.25%, higher than that of adults (1.56%) with 6-9 year group the highest (2.83% ). The morbidity of boys was 3.04% higher than that of girls (1.37%). Among the 7 death cases, 3 were caused by injury(2 RTA, 1 drowning). The three leading causes of injuries were fall, animal bite and road traffic accident in children. Among children aged 0-5 years, fall, animal and burn/machine injuries were the leading causes. Among children aged 0-5 years, the leading causes were hurt by animals, fall and RTA with the leading causes as fall, sprain and RTA. The average medical cost of injury cases was 1617.57 Yuan. There were 3500 children (injured orphan) whose parents were injured leading to death or disability in Beijing in 2003.Injury was the leading cause of child deaths in Beijing with children of 6-9 years old having the highest morbidity with RTA the most severe type of injuries in Beijing which affects both children and their parents.

To explore the efficacy and safety of the ultrathin flap LASIK and LASEK for the treatment of high myopia with central corneal thickness below 500 microm.Ultrathin flap LASIK and LASEK were randomly selected to treat high myopic patients with corneal thickness between 450 and 500 microm. 39 cases of 23 patients with average spherical equivalent of -7.51D (range from -6.00 to -9.50D) were treated with ultra thin flap LASIK, and 37 cases of 19 patients with average SE of -7.50D (range from -6.00 to -10.75D) were treated with LASEK. The uncorrected visual acuity, best corrected visual acuity, spherical equivalent, intraocular pressure, haze were examined and recorded 1, 3, 6 and 12 months postoperatively.Postoperative reaction was mild in LASIK patients, who showed a quick recovery in UCVA, whereas LASEK patients needed average 4 days to re-epithelization. At 1, 3, 6, 12 months postoperatively, the percentage of UCVA above 1.0 was 64.1%, 87.2%, 87.2% and 79.3% in LASIK patients and 37.8%, 75.7%, 67.6% and 71.4% in LASEK patients respectively. The percentage of spherical equivalent between +/- 0.50D was 48.7%, 51.3%, 61.5%, 82.8% in LASIK patients and 51.4%, 45.9%, 45.9%, 57.1% in LASEK respectively. There was no severe complications that implicate the BSCVA occurred in-operation and postoperatively. The main complications in LASIK group was corneal flap striae and overcorrection, regression and mild haze in LASEK group.Ultrathin flap LASIK had the similar efficacy and safety with LASEK in the treatment of high myopia with thin CCT, but the stability of results and satisfaction from the patients was superior to LASEK.

To compare the effectiveness and difference between laser subepithelial keratomileusis (LASEK) and conventional photorefractive keratectomy (PRK) for the treatment of myopia up to -8.00 diopter.In this prospective study, 46 patients with a manifest refactiion of -1.75 to -8.00 diopters were treated and followed up for 6 months. In each case, PRK was performed in one eye and LASEK in the other eye. The first eye treated and surgical method used in the first eye was randomized. Epithelial healing time, postoperative pain, uncorrected visual acuity (UCVA), manifest refraction, corneal HAZE were followed and compared in PRK and LASEK treated eyes.LASEK eyes took a mean time of 3.49 days to heal the epithelium, whereas PRK eyes took 2.45 days, which is statistically significant (P0.05). Postoperative pain index was 2.04 and 2.45 in LASEK eyes and PRK eyes respectively (P0.05). There were no significant differences between eyes in UCVA and manifest refraction during the follow time (P0.05). However, LASEK-treated eyes showed less corneal haze than PRK eyes.LASEK can effectively and safely treat myopia up to -8.00 diopters as PRK did, and can diminish early pain after surgery and prevent corneal Haze from happening.

To study the inhibitory effect of retroviral packaging cells injected intrasplenically encoding mouse interleukin-12 (mIL-12) and human interleukin-2 (hIL-2) fusion gene on the growth of hepatocellular carcinoma.The retroviral vectors encoding mIL-12 gene, hIL-2 gene, and mIL-12 and hIL-2 genes, GCIL12EXPN, GCXEIL2PN, and GCIL12EIL2PN were constructed and then transfected into the retroviral packaging cells PA317 to construct cells PA317-GCIL12EXPN, PA317-GCXEIL2PN, and PA317-GCIL12EIL2PN. Rat hepatocellular carcinoma cells CBRH3 were implanted into the livers of Wistar rats to establish hepatoma animal model. Then the rats were divided into 5 groups to be injected intrasplenically with normal saline one day after the implantation (0.8 ml/rat, group I, n = 10), blank vector PA317-GCXEXPN one day after the implantation (10(7) cells/rat, group II, n = 10), PA317-GCIL12EXPN containing IL-2 gene (1 x 10(7) cells/rat 1, 3, 5, or 7 days after the implantation, group III, n = 40), PA317-GCXEIL2PN containing mIL-12 gene (1 x 10(7) cells/rat 1, 3, 5, or 7 days after the implantation, group IV, n = 40), and PA317-GCIL12EIL2PN containing IL-12-IL-2 fusion gene (1 x 10(7) cells/rat 1, 3, 5, or 7 days after the implantation, group V, n = 40) respectively. The rats surviving longer than 2 months were re-injected with hepatocellular carcinoma cells. The therapeutic effect, immune function and toxic effect were evaluated. CT was conducted on the liver before and after the experiment. Laparotomy was performed 3 and 7 days after treatment to resect some of the carcinoma tissues to undergo pathological examination and OX8 immunohistostaining. Serum mIL-12 and hIL-2 were detected one day before and 3, 7, 30, and 60 days after treatment.The average survival times of the rats treated with IL-12-IL-2 fusion gene at the first, third, fifth and seventh day after tumor implantation were 53.3 +/- 3.7 days, 49.3 +/- 4.2 days, 31.0 +/- 2.1 days, and 24.3 +/- 1.4 days respectively, longer than those treated with IL-2 gene (25.0 +/- 2.5 days, 23.5 +/- 2.0 days, 18.3 +/- 2.4 days, and 12.0 +/- 1.8 days respectively, P0.001), and those treated with IL-12 gene (39.0 +/- 4.8 days, 32.0 +/- 3.9 days, 23.0 +/- 2.5 days, and 19.4 +/- 2.1 days respectively, P0.001). Long survival (or= 60 days) rate in the rats treated with IL-12-IL-2 fusion gene on the first and third day was 30%. The serum mIL-12 and hIL-2 levels in these rats remained high on the 60th day after treatment. The pathological study showed that the number of infiltrating lymphocytes in liver tumor tissues was increased in the IL-12-IL-2 fusion gene treatment group.The retroviral packaging cell line injected intrasplenically encoding mIL-12 and hIL-2 fusion gene inhibits the growth of hepatocellular carcinoma significantly in rats. The therapeutical efficacy of early administration is superior to that of late one.

AIM: To test the efficacy of gene therapy in rat liver tumor. METHODS: A retroviral vector GCIL12EIL2PN encoding human IL-2 (hIL-2) and mouse IL-12 (mIL-12) fused gene and its packaging cell were constructed. The packaging cell lines contained of IL-2 and/or IL-12 genes were injected intrasplenically to transfect splenocyte at different time. The therapeutic effect, immune function and toxic effect were evaluated. RESULTS: The average survival times of the 4 groups using IL genes at days 1, 3, 5 and 7 after tumor implantation were 53.3±3.7, 49.3±4.2, 31.0±2.1 and 24.3±1.4 d respectively in IL-2/IL-12 fused gene group, 25.0±2.5, 23.5±2.0, 18.3±2.4 and 12.0±1.8 d respectively in IL-2 gene treatment group, and 39.0±4.8, 32.0±3.9, 23.0±2.5 and 19.4±2.1 d respectively in IL-12 gene treatment group (P

AIM: The imaging features of MRI and DSA, using the models of implanted and induced hepatoma, were investigated in rats. METHODS: CBRH3 cancer cells were implanted for different liver site of rat liver and the diethylnitrosoamine was given orally to rats in order to induce liver cancer. Both experimental groups were detected by magnetic resonance imaging (MRI), digital subtraction angiography (DSA) and morphologic assay. RESULTS: Hypointensity on T1WI and homogenous high signal intensity on T2WI in MRI, and ring-like abnormal stain on DSA were found in implanted cancer. Induced cancers appeared as homogeneous or heterogeneous hypointensity on T1WI (10 cases), and equal or slight high intensity on T2WI (8 cases), but some as hypointensity on T2WI (2 cases). CONCLUSION: The imaging features of implanted cancers were similar to that of human liver metastases. Therefore, it could serve as an experimental model of human liver metastatic tumor. The imaging feature of induced cancers, whereas, were similar to that of human primary liver cancer. It could be use as an experimental model of human primary liver cancer.