Objective:To study the relationship between alpha seine/threonine-protein kinase (p-Akt)-serine/ threonine-protein kinase (mTOR)-ribosomal protein S6 kinase (p70S6K) signaling pathway and clinicopathological features or chemoresistance of ovarian cancer.Methods:We checked the p-Akt,mTOR and p70S6K protein levels in 18 tissues with chemoresistance or 25 with chemosensitivity of ovarian cancer by immunohistochemistry technique,and analyzed the relationship between those proteins and clinicopathological features or chemoresistance of ovarian cancer.Results:The levels of p-Akt protein in ovarian serous carcinoma,mucinous carcinoma and endometrioid carcinoma were 77.14%,50.00% and 66.67%,respectively,with no significant difference (P>0.05).The levels of these proteins in well-middle differentiated carcinoma and low differentiated carcinoma were 73.33% and 75.00%,respectively,with no significant difference (P>0.05).The levels of these proteins in Ⅰ-Ⅱ stage carcinoma,and Ⅲ-Ⅳ stage carcinoma were 18.18% and 93.75%,respectively,with significant difference (P<0.05).The levels ofmTOR protein in ovarian serous carcinoma,mucinous carcinoma and endometrioid carcinoma were 77.14%,100.00% and 83.33%,respectively,with no significant difference (P>0.05).The levels of this protein in well-middle differentiated carcinoma and low differentiated carcinoma were 80.00% and 78.57%,respectively,with no significant difference (P>0.05).The levels of this protein in Ⅰ-Ⅱ stage carcinoma,and Ⅲ-Ⅳ stage carcinoma were 27.27% and 96.88%,respectively,with significant difference (P<0.05).The levels of p70S6K protein in ovarian serous carcinoma,mucinous carcinoma and endometrioid carcinoma were 80.00%,100.00% and 100.00%,respectively,with no significant difference (P>0.05).The levels of this protein in well-middle differentiated carcinoma and low differentiated carcinoma were 93.33% and 78.57%,respectively,with no significant difference (P>0.05).The levels of this protein in Ⅰ-Ⅱ stage carcinoma,and Ⅲ-Ⅳ stage carcinoma were 45.45% and 96.88%,respectively,with significant difference (P<0.05).The levels of p-Akt protein in tissue of chemoresistance and chemosensitivity of ovarian cancer were 88.89% and 64.00%,respectively,with significant difference (P<0.05).The levels of mTOR protein in tissue of chemoresistance and chemosensitivity of ovarian cancer were 94.44% and 68.00%,respectively,with significant difference (P<0.05).The levels of p70S6K protein in tissue of chemoresistance and chemosensitivity of ovarian cancer were 100.00% and 72.00%,respectively,with significant difference (P<0.05).Conclusion:The p-Akt-mTOR-p70S6K signaling pathway may take part in invasion and metastasis of ovarian cancer.The up-regulation of these proteins may be associated with the chemoresistance of ovarian cancer,and these proteins may have potential to be the prognostic markers for the chemoresistance of ovarian cancer.

OBJECTIVE: To investigate the prognosis-related influence factors of the residual symptoms after the canalith repositioning procedure (CRP) for the benign paroxysmal positional vertigo (BPPV) in the second affiliated hospital of dalian medical university. METHOD: Among patients who were diagnosed with BPPV and treated by CRP, the one that still show residual symptoms were enrolled in our study, then make a follow-up irregularly about the tendency of their residual symptoms' self-healing,and respectively record in their gender, age and chronic diseases and so on. Single-factor analysis and multi-factors analysis was utilized to investigate the residual symptoms' related influencing factors. RESULT: In this study, 149 cases of patients were in record, for the residual symptoms, 71 patients can go to self-healing, 78 patients can not; age is 23-88, 30 cases in the young group, 46 cases in the middle aged group, 47 cases in the young elderly group, 26 cases in the elderly group; patients suffering from high blood pressure are 76 cases, 76 cases had diabetes, 47 cases had hyperlipidemia, 110 cases had heart disease, 43 cases had ischemic encephalopathy. CONCLUSION The residual symptoms in the elderly females patients and patients suffering from the hypertension, diabetes, heart disease patients and ischemic encephalopathy are not easy to heal by itself, in which, the older and the fact suffering from the hypertension and diabetes are the risk factors influencing the prognosis of the residual symptoms.
Adult ; Aged ; Aged, 80 and over ; Benign Paroxysmal Positional Vertigo ; complications ; therapy ; Chronic Disease ; Diabetes Mellitus ; Female ; Heart Diseases ; complications ; Humans ; Hypertension ; complications ; Male ; Middle Aged ; Patient Positioning ; Prognosis ; Risk Factors ; Young Adult

AIM:In podocytes , autophagy occurs at a high basal level and dysregulated autophagy is associa -ted with a variety of podocytopathies .This paper is to investigate the role of autophagy in sublytic C 5b-9-induced podocyte injury.METHODS: Sublytic complement C5b-9 stimulation was used as an in vitro model.Autophagosomes were con-firmed using monodansylcadaverine (MDC) staining.Immunoblotting was used to measure the change of autophagy-related markers.Cellular morphological changes were observed by Wright-Giemsa staining.Immunofluorescence staining and con-focal microscopy were used to detect the expression and distribution of nephrin .The cell viability was assessed by methylth-iazol tetrazolium (MTT) assay.The cell apoptosis was assessed by Annexin V-fluorescein isothiocyanate/PI staining.RE-SULTS:For ensuring sublytic complement injury , the maximal amounts of anti-podocyte antiserum and 160 ×-diluted nor-mal human serum were used without inducing cell lysis (defined as >5%LDH release).Sublytic C5b-9 promoted autoph-agy of podocytes in vitro.The proautophagic effect of sublytic C 5b-9 manifested in the form of accumulated MDC-labeled vesicles and enhanced the expression of LC 3-Ⅱ.Autophagy inhibitor 3-methyladenosine (3-MA) promoted sublytic C5b-9-induced podocyte morphological abnormalities .Compared with the sublytic C5b-9-injured podocytes, 3-MA exposure further decreased the expression of nephrin .3-MA enhanced sublytic C5b-9-induced podocyte apoptosis .CONCLUSION: Sub-lytic C5b-9 attack induces autophagy , which may play a protective role against complement-mediated podocyte injury .

Objective To analyze the risk factors associated with acute renal failure (ARF) in critically ill obstetric patients in intensive care unit (ICU).Methods A retrospective analysis of 192 critically ill obstetric patients admitted to the ICU for postpartum complications was conducted. There were 10 patients excluded because 3 died of amniotic embolism with massive hemorrhage in ICU and 7 had incomplete records, therefore, only 182 critically ill obstetric patients were enrolled in this study. According to the occurrence of ARF or not, the patients were divided into ARF group (68 cases) and non-ARF group (114 cases). The indicators of preeclampsia, HELLP syndrome (hemolysis, elevated liver enzymes, thrombocytopenia), acute fatty pregnancy liver, postpartum hemorrhage, sepsis were collected in the two groups. The main drugs used in the 4 days after delivery were as follows: ① plasma volume expanders: fluids (crystalloids, artificial colloids, hypertonic albumin and 4% serum albumin) and blood products (packed red cells, concentrated platelets, fibrinogen, fresh frozen plasma and activated factor Ⅶ); ② antifibrinolytic drug: tranexamic acid; ③ anti-hypertensive drugs: angiotensin-converting enzyme inhibitors (ACEI), diuretics; ④ nephrotoxic drugs: aminoglycosides, contrast agent. The above indexes were firstly analyzed by the univariate method, and the risk factors with statistical significance were further analyzed by multivariate logistic regression method to screen out the independent risk factors of developing ARF in critically ill obstetric patients in ICU.Results Univariate analysis showed that HELLP syndrome, tranexamic acid and hypertonic albumin infusion used in 4 days after delivery were the risk factors of ARF (χ2 value was 4.92, 4.29, 5.53, respectively, allP < 0.05). The multivariate logistic regression analysis showed: HELLP syndrome [odds ratio (OR) = 10.478, 95% confidence interval (95%CI) was 1.248 - 17.953,P = 0.030] and hypertonic albumin infusion used in 4 days after delivery [OR = 6.632, 95%CI was 1.211 - 16.328,P = 0.029] were the independent risk factors to develop ARF.Conclusions The occurrence of ARF in a critically ill obstetric patient in ICU is a process involving multiple factors, therefore, it is necessary to fully recognize the risk factors influencing the development of this disease; the presence of HELLP syndrome and hypertonic albumin infusion used in 4 days after delivery are the independent risk factors of developing ARF. Thus, particular attention should be paid to those patients with such high risk factors to decrease the incidence of ARF.

Objective To investigate the role of autophagy in podocyte damage,and the intracellular mechanism of autophagy activation through passive Heymann nephritis (PHN) animal model.Methods Male Sprague-Dawley rats (n=40) were studied on day 0,2,4,7,and 21 after induction of PHN by injection of anti-Fx1A.Podocyte morphology and autophagosomes were observed by transmission electron microscopy.Podocyte numerical density was estimated by Weibel-Gomez =method.Cell apoptosis was detected by TUNEL assay and caspase-3 immunohistochemical staining.Expressions of autophagy markers and endoplasmic reticulum stress (ERS)-associated proteins were analyzed by Western blotting.Results (1) In PHN rats,immunohistochemical staining showed that C5b-9 deposited along glomerular basement membrane on day 4 to day 21.Small subepithelial electron -dense deposits and a part of foot process fusion were detected in the glomerulus of PHN rats on day 4 by transmission electron microscope,and podocyte damage was aggravated on day 21.Furthermore,compared with control,the urinary protein levels of PHN rats began to increase on day 3,and reached the top on day 21 [(50.6±6.0) mg/24 h].(2) The number of podocytes significantly decreased in PHN rats compared with control group on day 21(P ＜ 0.05).(3) In PHN rats,apoptotic podocytes were found by caspase-3 immunohistochemical staining and TUNEL assay on day 21.(4) The expression of autophagy marker LC3 Ⅱ was markedly increased on day 7 and 21,but down-regulated on day 21 compared with day 7.Moreover,accumulated autophagosomes in podocytes were detected on day 7 and 21 by transmission electron microscope.(5) The level of GRP78 was significantly increased on day 2 and 7 but reduced to baseline on day 21.At the same time,the downstream pathways (ATF6α,p-PERK and p-JNK) of unfolded protein response were also up-regulated in the early process of PHN and down-regulated later.Conclusions Autophagy is an important way to protect against immunemediated podocyte injury in membranous nephropathy.Autophagy activation is mainly related to endoplasmic reticulum stress induced by complement attack.This provides an important basis for a thorough understanding of the role of autophagy in the process of podocyte damage and the pathogenesis of membranous nephropathy.

AIM: To determine the effect of rapamycin on the progression of passive Heymann nephritis (PHN), and whether autophagy is involved in this process .METHODS:Male Sprague-Dawley rats (n=24) were ran-domly divided into 3 groups:control group , PHN group and rapamycin treatment group .The rat PHN model was induced by injection of anti-Fx1A serum through penile vein , and all rats were sacrificed on day 21.Automatic biochemical analyzer was used to detect 24 h urine protein , blood urea nitrogen and serum creatinine .Renal damage was observed through per-iodic acid-silver methenamine staining .The number of podocyte was estimated by Weibel-Gomez method .The glomerular deposition of C5b-9, the expression of caspase-3 and expression of autophagy marker LC 3 in glomeruli were examined by immunofluorescence staining , immunohistochemical staining and Western blotting , respectively.RESULTS: Rapamycin significantly reduced proteinuria in the PHN rats (P<0.05), while the renal functions in 3 groups were normal, without significant difference .Although rapamycin limited weight gain in the rats , the health of the rats during drug treatment was not affected .Rapamycin retarded glomerular basement membrane thickening in the PHN rats .Rapamycin significantly re-duced the podocyte deletion by preventing podocyte apoptosis .Rapamycin enhanced the level of autophagy of glomerular in-herent cells .CONCLUSION:In the disease process of PHN , appropriate strength of autophagy plays a protective role . Rapamycin appropriately enhances autophagy and prevents podocyte apoptosis , thus reducing nephropathy and proteinuria . This may be one of the important mechanisms of rapamycin to slow down the progress of PHN .

Objectives Construct a subtractive library of Caski cell line induced by exposing to the space environment by suppression subtractive hybridization and pave the way to explain the molecular mechanisms of the changes at the gene level. Methods Super SMART cDNA synthesis and suppression subtractive hybridization (SSH) were performed to isolate differentially expressed cDNA fragments from strains subclonal 48A9 cell line. cDNA from the 48A9 cell line were used as " tester" , and the other from the control Caski cell line as "driver". Subtractive products were directly inserted into T/A cloning vector, and then transformed into host bacteria to set up a subtractive cDNA library of specially or highly expressed genes in strains subclonal 48A9 cell line. Results mRNA were directly extracted and purified with good quality. Double strand cDNA were reverse transcripted integratedly, and then cut by Rsa I into even length short segments. Liga-tion was identified as high effective. After two hybridizations, a subtractive library of differentially expressed genes in strains subclonal 48A9 cell line was successfully constructed by SSH. Conclusion SSH is an effective approach to isolate differentially expressed genes.

Objective To determine the effect of rapamycin on sublytic C5b-9-induced podocyte adhesion damage,and whether autophagy is involved in this progression.Methods Sublytic complement C5b-9 stimulation was used in vitro.Autophagosomes were viewed using electron microscopy.Western blotting was used to measure the change of autophagy-related markers.Attachment assay was used to assess the adhesion of podocyte.Confocal microscopy was used to explore the expression patterns of cytoskeletal protein F-actin.Flow cytometry was used to measure the level of adhesion-associated protein integrin α3.Results (1) For ensuring sublytic complement injury,the maximal amounts of anti-podocyte antiserum and 160×-diluted normal human serum were used without inducing cell lysis (defined as ＞ 5％ LDH release).(2) Sublytic C5b-9 promoted autophagy in podocyte in vitro.The proautophagic effect of sublytic C5b-9 manifested in the form of accumulated autophagosomes and enhanced expression of LC3-lⅡ.(3) Inhibition of autophagy by 3-methyadenine enhanced the effect of sublytic C5b-9-induced podocyte injury,including serious cytoskeleton damage and markedly reduced adhesion of podocyte.(4) Rapamycin treatment significantly improved the above lesions.(5) Rapamycin enhanced autophagy induced by sublytic C5b-9 in podocyte.Conclusions In summary,rapamycin can improve sublytic CSb-9-induced podocyte adhesion damage by appropriate autophagy activation.These findings provide important information for the development of appropriate protocols for the application of mTOR (mammalian target of rapamycin) inhibitors in podocytopathy.

Objective To investigate the relationship between baseline serum uric acid and the severity of coronary artery disease ( CAD ) in the first-degree relatives or non-first-degree relatives of men with type 2 diabetes. Methods Three hundred and eighty-one men with negative coronary angiography for the first time were divided into diabetes and non-diabetes groups and followed-up for 5 years. The primary outcome was acute coronary syndrome suspected during subsequent 5 years, and the coronary angiography was conducted simultaneously. The severity of CAD was assessed by the coronary stenosis index ( CSI) and the number of coronary lesion vessels. Results In normal blood glucose group, serum uric acid was higher in the first-degree relatives of diabetics compared with non-first-degree relatives(P<0. 01), along with higher morbidity of CAD, CSI, and coronary lesion vessels (all P<0.01). Correlation analysis showed that CSI(r=0. 250, P=0. 041) and coronary lesion vessels(r=0. 252, P=0. 040) in non-diabetics group were associated with baseline levels of serum uric acid. Conclusion The elevation of serum uric acid was closely related to subsequent CAD, especially in first-degree relatives of male with type 2 diabetes, which could be used as an early indicator for CAD prediction.

Aim Todiscusstheapplicationoftissue spontaneous fluorescence in myocardial ischemia injury of rats based on specific fluorescence detection technol-ogy.Methods Thechangeofspontaneousfluores-cence was compared between the myocardial tissue of normal rats and those of rats with myocardial ischemia injury and an quantitative analysis was then made.Re-sult Theresultsshowedthatspontaneousfluorescence of myocardial tissue for myocardial ischemia injury changed significantly.Spontaneous fluorescence signal of injury considerably was enhanced.The fluorescence signal which was quantified by FX Pro had statistical significance compared with normal myocardial tissue,P<0.01orP<0.05.Conclusion Ourresearchshows that spontaneous fluorescence of myocardial tissue can be enhanced obviously after myocardial ischemia inju-ry.Our research provides a method for the research and evaluation of myocardial ischemia injury model in rats which can be used in positioning,qualitative and quan-titative comparative study and in pathological,physio-logical,pharmacological and mechanism study.