Overexpression of chaperonin containing TCP1, subunit 3 predicts poor prognosis in hepatocellular carcinoma

AIM:To investigate the value of chaperonin containing TCP1,subunit 3(CCT3) to predict the prognosis of patients with hepatocellular carcinoma(HCC) and determine its function in HCC progression.METHODS:CCT3 expression levels were examined in human non-cancerous liver tissues and a variety of HCC cell lines by quantitative real-time PCR and immunoblotting.CCT3 expression was suppressed by small interfering RNA.The effects of reducing CCT3 expression in HCC cells were tested.The3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide(MTT) assay,cell counting experiment,cell cycle assay,apoptosis assay and invasion assay were employed to evaluate cell functions in vitro.Immunohistochemistry was performed on HCC specimens.In addition,CCT3 expression in HCC specimens was also assessed at the protein and mRNA level.Associations between clinicopathological characteristics and prognosis were analyzed,along with the possible mechanisms involved in CCT3's function in HCC progression.RESULTS:The expression levels of CCT3 mRNA and protein were upregulated in HCC cell lines in contrast to adjacent non-cancerous tissues.Reducing CCT3 expression not only suppressed cell proliferation in cell counts,MTT assay,cell cycle assay and induced cell apoptosis(P < 0.05 vs negative control),but also inhibited the tumor cell invasion capacity in vitro {P< 0.01 vs negative control).Overexpression of CCT3 in the nuclei of cancer cells in HCC specimens(58of 104 patients,55.8%) was associated with poor prognosis in HCC patients(3-year survival rate,55.5%vs 84.2%,P = 0.020) after hepatectomy.Mechanistic analyses showed that signal transducer and activator of transcription 3(STAT3) activation was decreased even when stimulated by interleukin-6 after knocking down CCT3 in the HepG2 cell line.CONCLUSION:Overexpression of CCT3 in the nuclei of cancerous cells is associated with HCC progression.CCT3 may be a target that affects the activation of STAT3 in HCC.

Neuroprotective effects of atorvastatin against cerebral ischemia/reperfusion injury through the inhibition of endoplasmic reticulum stress

Cerebral ischemia triggers secondary ischemia/reperfusion injury and endoplasmic reticulum stress initiates cell apoptosis. However, the regulatory mechanism of the signaling pathway remains unclear. We hypothesize that the regulatory mechanisms are mediated by the protein kinase-like endoplasmic reticulum kinase/eukaryotic initiation factor 2α in the endoplasmic reticulum stress signaling pathway. To verify this hypothesis, we occluded the middle cerebral artery in rats to establish focal cerebral ischemia/reperfusion model. Results showed that the expression levels of protein kinase-like endoplasmic reticulum kinase and caspase-3, as well as the phosphorylation of eukaryotic initiation factor 2α, were increased after ischemia/reperfusion. Administration of atorvastatin decreased the expression of protein kinase-like endoplasmic reticulum kinase, caspase-3 and phosphorylated eukaryotic initiation factor 2α, reduced the infarct volume and improved ultrastructure in the rat brain. After salubrinal, the specific inhibitor of phosphorylated eukaryotic initiation factor 2α was given into the rats intragastrically, the expression levels of caspase-3 and phosphorylated eukaryotic initiation factor 2α in the were decreased, a reduction of the infarct volume and less ultrastructural damage were observed than the untreated, ischemic brain. However, salubrinal had no impact on the expression of protein kinase-like endoplasmic reticulum kinase. Experimental findings indicate that atorvastatin inhibits endoplasmic reticulum stress and exerts neuroprotective effects. The underlying mechanisms of attenuating ischemia/reperfusion injury are associated with the protein kinase-like endoplasmic reticulum kinase/eukaryotic initiation factor 2α/caspase-3 pathway.

The key target of neuroprotection after the onset of ischemic stroke: secretory pathway Ca^(2+)-ATPase 1

The regulatory mechanisms of cytoplasmic Ca2＋ after myocardial infarction-induced Ca2＋ overload involve secretory pathway Ca2＋-ATPase 1 and the Golgi apparatus and are well understood. However, the effect of Golgi apparatus on Ca2＋ overload after cerebral ischemia and reperfusion remains unclear. Four-vessel occlusion rats were used as animal models of cerebral ischemia. The expression of secretory pathway Ca2＋-ATPase 1 in the cortex and hippocampus was detected by immunoblotting, and Ca2＋ concentrations in the cytoplasm and Golgi vesicles were determined. Results showed an overload of cytoplasmic Ca2＋ during ischemia and reperfusion that reached a peak after reperfusion. Levels of Golgi Ca2＋ showed an opposite effect. The expression of Golgi-specific secretory pathway Ca2＋-ATPase 1 in the cortex and hippocampus decreased before ischemia and reperfusion, and increased after reperfusion for 6 hours. This variation was similar to the alteration of calcium in separated Golgi vesicles. These results indicate that the Golgi apparatus participates in the formation and alleviation of calcium overload, and that secretory pathway Ca2＋-ATPase 1 tightly responds to ischemia and reperfusion in nerve cells. Thus, we concluded that secretory pathway Ca2＋-ATPase 1 plays an essential role in cytosolic calcium regulation and its expression can be used as a marker of Golgi stress, responding to cerebral ischemia and reperfusion. The secretory pathway Ca2＋-ATPase 1 can be an important neuroprotective target of ischemic stroke.

Effects of Growth Hormone Supplementation in Patients Undergoing IVF/ICSI-ET with Poor Ovarian Response to Gonadotropin

Objective To analyze the effects of growth hormone （GH） supplementation during IVF/ ICSI-ET in Chinese patients who had prior IVF cycle with poor response to gonadot- ropin （Gn）. Methods Ovulation was stimulated in 389 consecutive patients who all had poor ovarian response, among them, 102 patients （GH cycle） received 4 IU GH and the other 287 patients （non-GH cycle） underwent IVF without GH. Fisher＇s exact test, Chi square test and Student＇s t-test were used to analyze IVF/ICSI-ET outcomes. Results After GH treatment, 102 patients had significantly more large- and medium- sized follicles, oocytes retrieved, 2 pronucleus oocytes, metaphase H stage （M^I） oocytes, and high-quality embryos than in previous cycles without GH. However, the number of embryos transferred, clinical pregnancy rate, transfer rate and biochemical pregnancy rate were not significantly different. Furthermore, the 102 patients given GH had significantly lower luteinizing hormone levels and biochemical pregnancy rates; thicker endometrium and more Gn administration days; and more large- and medium-sized follicles and M^I oocytes than 287 other patients undergoing IVF/ICSI-ET without GH. However, these groups did not differ significantly in clinical pregnancies, high-quality embryos, Mn oocytes, and embryo implantation rates. Conclusion GH may improve some IVF/ICSI-ET outcomes for women with poor ovarian response.

Study on the Differences of Endocrine and Metabolic Characteristics in Clinical Subtypes of PCOS

Objective To investigate the differences of endocrine and metabolic characteristics in PCOS women among different subtypes. Methods A total of 249 PCOS women were classified into 4 subtypes by Rotterdam criteria： hyperandrogenism （HA） and oligo-ovulation/anovulation （0） and PCO ultrasonography （P） （HA＋O＋P, group A, 111 women）; HA＋O （group B, 9 women）; HA＋P （group C, 22 women） and O＋P （group D, 107 women）. Another 110 infertii＇e women with tubal defects constituted a control group. Endocrine and metabolic characteristics were compared among the 5 groups. PCOS women were then reclassified into 2 groups, according to whether they have hyperandrogenism or not. Endocrine ond metabolic characteristics were then compared again. Results The levels of androstenedione （A）, testosterone （T） and LH/FSH were the highest in group A and group C, secondly in group D, the lowest was in control group. A, T and LH/FSH were the highest in hyperandrogenism group, secondly in non- hyperandrogenism group, whereas the control was the lowest. Menstrual cycle and BMI correlated with glucose and lipid metabolism but showed no correlateion with T and A. Hyperandrogenism group had higher fasting glucose （FG）, glucose at 60 min （G60） and glucose levels under the curve （GLUAUC） and lower disposition index （DI） than non-hyperandrogenism group, however, menstrual cycle, BMI and the lipid indicators had no difference. Conclusion Hyperandrogenism is an important characteristic in F＇COS women. Thus it might be used to classify PCOS into 2 subgroups. Hyperandrogenism and lipid disorders for the formation of PCOS often coexist but each has individual pathogenesis.

Study on the Clinical and Endocrine Characteristics of Polycystic Ovary Syndrome with Different Ovarian Morphology

Objective To evaluate the differences of the clinical manifestation and endocrine situation in patients with different ovarian morphology of polycystic ovary syndrome （PCOS）. Methods A total of 234 PCOS patients were enrolled according to the ovary morphology and divided into three groups： 112 patients with B-polycystic ovary mor- phology （both two ovaries were PCOM, B-PCOM）, 50 with U-PCOM （only one ovary was PCOM） and 72 with N-PCOM （none was PCOM）. There were 39 infertile women without PCOS as control group. Data were analyzed by using SPSS 15.0 software. ResuIts There was no statistical difference in body mass index （BM1） among the three groups of PCOS. The endometrial thickness increased in patients with B-PCOM and decreased with N-PCOM. The levels of testosterone, androstenedione and luteinizing hormone increased in PCOS groups, especially in N-PCOM patients. HOMA-IR increased, HOMA-fl, disposition index （DI） and △160/AG60 decreased in patients with N- PCOM compared with in B-PCOM and U-PCOM groups. Higher level of total choles- terol （TC） and lower level of high-density lipoprotein （HDL）-C existed in PCOS patients, especially in N-PCOM. There were positive correlations between oligo-anovulation, endometrial thickness, LH/FSH ratio, fasting insulin （FINS）, the area under curve of glucose（A UCcLu） and PCOM, while there was a negative correlation between HOMA- IR and PCOM. Conclusion There are relationships among hyperandrogenism, hyperinsulinemia, insulin resistance （IR） and ovary morphology in PCOS patients. PCOS patients with- out PCOM have more serious IR and hyperandrogenism.

Posterior reversible encephalopathy syndrome in a woman with pancreatitis

To the Editor:Posterior reversible encephalopathy syndrome (PRES) is a rare condition of the central nervous system (CNS).Clinically,it can present with headache,blurred vision,seizures,or coma.The diagnosis of PRES is based on the classic radiologic imaging studies at occipital and parietal lobes in the subcortical white matter,and sometimes in the cortex.Hypertensive encephalopathy,eclampsia,immunosuppressive agents,and cytotoxic drugs can cause PRES.A patient with PRES may recover without sequela after removal of the causative factors.Notably,uncertainty in diagnosis and delay in treatment would have probably aggravated CNS injuries or death.However,the underlying mechanism of the disease remains unclear and controversial.

Single Nucleotide Polymorphisms (SNPs) and Variable Number Tandem Repeats (VNTRs) in mtDNA D-loop and CO Ⅱ-tRNA^(Lys) Intergenic Region with PCOS

Objective To explore the relationships intergenic region in mtDNA with PCOS. of variations in D-loop and COH-tRNA^Lys Methods A total of 77 PCOS and 45 non-PCOS patients were enrolled, whose D-loop and COH-tRNA^Lys intergenic region in mtDNA were amplified and sequenced; sexual hormone assay, oral glucose tolerance test （OGTT） and insulin releasing test were carried out. Then variations found in mtDNA were compared between the two groups, the correlations between variations and clinical indexes were analyzed in all subjects. Results Nucleotide variations found in mtDNA were not different between the two groups, but the mutation of 16 094T/C was found associated with the serum levels ofT and fasting insulin; （303-317）CnTC, associated with the serum levels of A and LH; 195C/T with A level and 491T/C with LH level; （8 272-8 289）（ACCCCCTCT）, was associated with the serum level of 1 h glucose. Conclusion Noncoding region mutations in mtDNA perhaps associate with PCOS clinical symptoms and involve in PCOS development.