[The diagnostic potential of Golgi protein 73 for cirrhosis in patients with chronic hepatitis C].

Objective: To explore the diagnostic value and model of serum Golgi protein 73 (GP73) in patients with hepatitis C cirrhosis. Methods: 271 cases with chronic hepatitis C virus infection who were treated in the Fifth Medical Center of PLA General Hospital from January 2010 to December 2017 were retrospectively collected as the research objects, including 126 cases with hepatitis and 145 cases with liver cirrhosis. Serum GP73 and liver stiffness measurement (LSM) based on transient elastography test were performed in all patients. Simultaneously, blood routine, liver function, coagulation function and other related indicators were collected. GP73 diagnostic efficiency for liver cirrhosis was evaluated by receiver operating characteristic curve (ROC). GP73 diagnostic value was clarified after comparison with aspartate aminotransferase/platelet ratio index (APRI), FIB-4 index (FIB-4) and LSM. Compensated hepatitis C virus-related cirrhosis diagnostic model based on serological index was established by logistic regression analysis. Results: The area under the receiver operating characteristic curve (AUC) of GP73, LSM, FIB-4 and APRI in the diagnosis of compensated hepatitis C virus-related cirrhosis were 0.923, 0.839, 0.836 and 0.800 respectively, and GP73 had the best diagnostic efficiency (P <0.001). LSM and GP73 combined use had improved the diagnostic sensitivity of cirrhosis to 97.24%. Multivariate logistic regression analysis revealed that GP73, age, and platelets were independent predictors of cirrhosis.Compensated hepatitis C virus-related cirrhosis diagnostic model (GAP) was established based on the result: LogitP=1/[1+exp(6.145+0.013×platelet-0.059×age-0.059×GP73)].AUC model for diagnosing compensated liver cirrhosis was 0.944, and the optimal cut-off value was 0.56, with sensitivity and specificity of 84.03% and 92.06%, respectively, and the diagnostic efficiency of this model was better than that of APRI, FIB-4, LSM and GP73 alone (P<0.05). Conclusion: GP73 is a reliable serum biomarker for the diagnosis of compensated hepatitis C virus-related cirrhosis. The GAP diagnostic model based on GP73, platelet count, and age can further improve the diagnostic efficiency and help to diagnose patients with compensated hepatitis C virus-related cirrhosis.

[Study on the prognostic value of preoperative anti hepatitis B virus therapy in patients with hepatocellular carcinoma complicated with microvascular tumor thrombus and establishment of survival prediction model].

Objective: To explored the effect of preoperative antiviral therapy on the prognosis of microvascular tumor thrombi patients, and to established a prognostic prediction model for these patients after radical resection of liver cancer. Methods: The clinicopathological and survival data of hepatocellular carcinoma patients with microvascular tumor thrombus who underwent radical resection in the Third Affiliated Hospital of Sun Yat-sen University from January 1, 2013 to December 31, 2015 were retrospectively collected. Kaplan-Meier method was used to calculate the survival curve, and log-rank test was used to compare the prognosis of patients with and without antiviral treatment before operation. Univariate and multivariate Cox proportional hazard regression model was used to screen predictive factors. R software was used to make predictive nomogram, and discrimination and calibration degree were used to evaluate the prediction model. Results: Among all 153 patients, 22 were female and 131 were male, aged (51.3±11.7) years. The preoperative antiviral therapy significantly improved overall survival and recurrence-free survival (χ2=41.423, 54.389; both P<0.001). According to the results of multivariate and regression analysis, preoperative antiviral therapy (HR=0.301,95%CI:0.171-0.532,P<0.001), alpha fetoprotein (HR=1.226,95%CI:1.157-1.776,P=0.032) and tumor size (HR=1.008,95%CI:1.001-1.016,P=0.02) were important prognostic factors for overall survival. The area under curve value of 3-year survival prediction model was 0.749(95%CI: 0.712-0.782), and that of 5-year survival prediction model was 0.755(95%CI: 0.724-0.793), with good calibration. Conclusions: Preoperative anti hepatitis B virus(HBV) therapy can significantly improve the prognosis of patients with hepatocellular carcinoma complicated with microvascular tumor thrombus, we develope the prediction models of 3-year and 5-year survival rate that can improve the reference for clinical work and benefit patients.

[Level and trend of respiratory diseases mortality in China from 2002 to 2016].

Objective: To analyze the level and trend of respiratory disease mortality in China from 2002 to 2016. Methods: The standardized mortality rates were calculated based on the China health statistics yearbook (2003-2012) and China statistical yearbook of health and family planning (2013-2017) data released by the statistical information center of National health Commission of the People's Republic of China. Joinpoint model was used to calculate the standardized mortality rates (SMR), Annual percentage change (APC) and the average annual percentage change (AAPC) for standardized mortality rates. Results: The SMR of respiratory diseases and chronic lower respiratory diseases were decreased significantly in 2002 to 2016 (AAPC=-3.6%, AAPC=-6.4%, P<0.001, respectively). The SMR of lung cancer showed a significant increase trend (AAPC=1.6%, P=0.001). There were no significant differences in the SMR of pneumonia and pneumonoconiosis (APCC=1.0%, P=0.242; APCC=-0.2%, P=0.905). Both urban and rural SMR of respiratory diseases were declining significantly (AAPC=-2.9%, P=0.001; AAPC=-4.2%, P<0.001). Both urban and rural SMR of lung cancer showed an increasing trend (AAPC=0.6%, P=0.022; AAPC=2.1%, P=0.003, respectively). The SMR of pneumonia in urban areas showed an upward trend (AAPC=2.7%, P=0.017). The SMR of respiratory disease of all age groups (<35 years old, 35-65 years old and ≥65 years old) showed a downward trend (AAPC=-3.8%, P=0.001; AAPC=-2.6%, P<0.001; AAPC=-3.9%, P<0.001). The SMR of pneumonia between 35 and 65 years old and SMR of lung cancer over 65 years old showed an increasing trend (AAPC=2.8%, P=0.001; AAPC=2.4%, P<0.001). The SMR of respiratory diseases among males and females showed a downtrend (AAPC=-3.1%, P<0.001; AAPC=-4.3%, P<0.001). However, the SMR of lung cancer in males and females increased significantly (AAPC=1.2%, P<0.001; AAPC=2.5%, P<0.001, respectively). There were no significant trends in the SMR of pneumonia and pneumoconiosis in males (AAPC=1.5%, P=0.096; AAPC=-1.6%, P=0.218). There was no obvious trend in the SMR of pneumonia in females (AAPC=-0.1%, P=0.872). Conclusions: The SMR of respiratory diseases in China generally shows a downward trend. The overall SMR and SMR of major respiratory diseases varies among different regions, genders and age groups.

[Establishment and preliminary application of serum Golgi protein 73 based noninvasive diagnostic model for compensated stage hepatitis B cirrhosis].

Objective: To establish and evaluate diagnostic efficacy and applicability of serum Golgi protein (GP) 73 based non-invasive diagnostic model with other conventional serological indicators for compensated stage hepatitis B cirrhosis. Methods: 666 cases with chronic hepatitis B (CHB) who had visited to the Fifth Medical Center of People's Liberation Army General Hospital from January 2010 to December 2017 were selected as the study subjects, and were classified according to compensated stage cirrhosis into clinical and pathological diagnosis group based on whether or not the liver histological examination was performed. A diagnostic model of compensated stage hepatitis B cirrhosis in the clinical diagnosis group was established. The current clinically used diagnostic model of liver cirrhosis, aspartate aminotransferase/platelet ratio index (APRI), fibrosis index (FIB)-4 and liver stiffness measurement (LSM) were compared. Eventually, the diagnostic model was verified step by step by pathological diagnosis group. Results: The area under the receiver operating characteristic curve (AUC) of GP73 and APRI, FIB-4, and LSM for cirrhosis patients in the clinical diagnosis group were 0.842, 0.857, 0.864, and 0.832, respectively. The diagnostic efficiency of the four indicators were of similar (P value > 0.05). A diagnostic model of compensated stage hepatitis B cirrhosis (GAPA) using logistic regression analysis was established: LogitP = 1/ [1 + exp (1.614-0.054 × GP73-0.045 × Age + 0.030 × PLT-0.015 × ALP)]. The AUC of the model was as high as 0.940 and the optimal cut-off value were 0.41. The corresponding diagnostic sensitivity and specificity were 0.92 and 0.82, respectively. The diagnostic efficiency was better than that of APRI, FIB-4, LSM and GP73 alone (P < 0.05). The AUC of GAPA was 0.877 in the pathological diagnosis group, which was similar to the diagnostic efficacy of LSM (0.891) and FIB-4 (0.847) (P > 0.1), but still superior to that of APRI (0.811) and GP73 alone (0.780) (P < 0.001). Conclusion: GAPA, a diagnostic model for compensated stage hepatitis B cirrhosis established in this study, has a good diagnostic efficacy in both the clinical and pathological diagnosis group, and has certain auxiliary diagnostic value in the areas where resources are relatively scarce or where LSM has not been developed.

[Prognostic significance of albumin/globulin ratio on postoperative survival of patients with hepatocellular carcinoma].

Objective: To investigate the prognostic value of albumin/globulin ratio on postoperative survival outcomes in patients with hepatocellular carcinoma. Methods: Data of 630 patients with HCC, who underwent surgical resection from February 2009 to July 2013, were retrospectively analyzed. Patients were divided into low-value group (A/G < 1.5, defined as L group) and high-value group (A/G≥1.5, defined as H group), and their distribution characteristics were observed with the normal A/G threshold value. Independent risk factors' affecting survival and prognosis was analyzed with univariate and multivariate Cox's regression model. Survival trend of all patients with low-value and high-value groups in A, B and C of Barcelona stage (BCLC stage) were analyzed using the Kaplan-Meier method. Results: Multivariate analysis showed that preoperative A/G ratio (P = 0.007), alpha-fetoprotein (P < 0.001), gamma-glutamyltransferase (P = 0.006), RBC (P = 0.014), international normalized ratio (P = 0.009), preoperative BCLC staging (P < 0.001) and number of tumors (P = 0.003), and intraoperative blood transfusion (P < 0.001) were independent prognostic factors affecting long-term survival in HCC patients. The median overall survival time in-group L was 15 months, significantly lower than that in group H of 42 months (P < 0.001). Stratified analysis showed that the short-term survival advantage of patients with high A / G value was limited to those with Barcelona stage A (P < 0.001), and disappeared in patients with Barcelona stage B and C (P > 0.05). The long-term survival advantage existed in patients with Barcelona stage A (P < 0.001), B (P < 0.05), and disappeared in C (P > 0.05). Conclusion: Preoperative albumin/globulin ratio can predict postoperative prognosis and survival, and direct towards the treatment for early stage of HCC and thus representing as an indicator of high clinical value.

α-SMA-Cre-mediated excision of PDK1 reveals an essential role of PDK1 in regulating morphology of cardiomyocyte and tumor progression in tissue microenvironment.

The phosphoinositide-3 kinase (PI3K) - phosphoinositide-dependent protein kinase 1 (PDK1)-Akt/protein kinase B (PKB) cascade plays a critical role in cardiovascular development and tumor genesis. But the role of PDK1 in the microenvironment of heart and tumor remains unknown. To clarify the effects of PDK1 on tissue microenvironment in vivo, here, we created α-SMA-Cre-mediated excision of PDK1 mice. And the mice were injected subcutaneously with Lewis lung carcinoma (LLC) cells. We found PDK1-deficient mice had post-natal praecox dilated cardiomyopathy, decelerated tumor growth and severe tumor metastasis. Histopathological analysis revealed abnormality of vascular microenvironment in heart and primary tumor. In conclusion, PDK1 plays a pivotal role in regulating cardiac function and tumor metastasis by interfering with microenvironment.

Effect of miR-124 on neuronal apoptosis in rats with cerebral infarction through Wnt/ß-catenin signaling pathway.

OBJECTIVE: The aim of this study was to observe the influence of micro ribonucleic acid (miR)-124 on neuronal apoptosis in rats with cerebral infarction (CI), and to further investigate the underlying mechanism of miR-124 in CI occurrence and development. MATERIALS AND METHODS: A total of 60 adult male Wistar rats were randomly divided into the Sham group, the CI group and the CI + miR-124 mimics group using a random number table. The focal CI model was established using the suture-occluded method. After successful modeling, miR-124 mimics were stereotactically injected into the lateral ventricle of rats. 24 h after operation, the neurological function of rats in each group was scored using modified neurological severity score (mNSS). Meanwhile, the infarction area of brain tissues was evaluated by the triphenyl tetrazolium chloride (TTC) method. The protein expression levels of apoptosis-related genes, including B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), C-Caspase and T-Caspase, were detected via Western blotting. The expression and location of caspase-3 in brain tissues were detected via immunofluorescence staining. Moreover, the level of apoptosis in each group was detected via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. In addition, the expression levels of the Wnt/ß-catenin signaling pathway-related proteins were detected via Western blotting. RESULTS: Polymerase Chain Reaction (PCR) results revealed that the expression level of miR-124 in the CI group was significantly decreased when compared with that of the Sham group (p<0.05). The mNSS and TTC staining results manifested that injection of miR-124 mimics could significantly reduce the CI-induced neurological deficits and CI area (p<0.05). At the same time, the levels of Bax and C-caspase/T-Caspase were significantly decreased, whereas the expression of Bcl-2 was remarkably increased after the injection of miR-124 mimics (p<0.05). Besides, the number of apoptotic cells in the CI + miR-124 mimic group was remarkably decreased (p<0.05). In addition, miR-124 mimics significantly activated the expression levels of Wnt and ß-catenin (p<0.05). CONCLUSIONS: The inhibitory effect of miR-124 on neuronal apoptosis in CI rats is probably related to the activation of the Wnt/ß-catenin signaling pathway. Furthermore, miR-124 is expected to be a target drug for the clinical treatment of CI.

[Acute effect of air pollutants (carbon monoxide and ozone) on myocardial infarction mortality in Ningbo].

Objective: To explore the effect of carbon monoxide (CO) and ozone (O(3)) in the air on the myocardial infarction mortality in Ningbo, Zhejiang province, from 2011 to 2015. Methods: The data of daily air quality surveillance and the causes of deaths in Ningbo from January 1, 2011 to December 31, 2015 were collected and the time series study using a generalized additive model was conducted to evaluate the relationship between the mortality of myocardial infarction and the air pollutants after adjustment for the long-term trend of death, weather conditions," days of the week" and other confounding factors. Results: The daily average concentrations of CO and O(3) in Ningbo during 2011-2015 were 0.90 (0.02-3.31) mg/m(3) and 82.78 (4-236) µg/m(3), respectively. A total of 5 388 myocardial infarction deaths occurred, with a daily average of 3 deaths. In single-pollutant model, an increase of 0.1 mg/m(3) in average concentration of CO could increase the risk of myocardial infarction mortality by 1.06% (95% CI: 0.29%-1.93%) in general population, and by 1.26% (95% CI:0.28%-2.24%) in aged people aged ≥65 years in lagged 6 days, but the influence was not significant in people aged <65 years. The influence had no significant difference in males, but it increased the risk of myocardial infarction mortality by 1.77% in females (95% CI: 0.44%-3.13%). In multi-pollutant model, CO did remain robust after adjusting for other co-pollutants. Whereas the effect of O(3) had no significant influence. Conclusion: These findings suggested that the increased risk of daily myocardial infarction mortality was associated with the increase of CO concentration, but no such association was found for O(3) in Ningbo.

[Relationship between particulate matters and cardio-cerebrovascular mortality in Ningbo: a time-series study].

OBJECTIVE: To explore the short-term effect of particulate matters with an aerodynamic diameter of less than or equal to 10 µg (PM10) and aerodynamic diameter of less than or equal to 2.5 µg (PM2.5) on cardio-cerebrovascular mortality in Ningbo city. METHODS: Daily cardio-cerebrovascular mortality data from 2011 to 2014 in Ningbo city were collected and the time series study using a semi-parametric generalized additive model were used to evaluate the relationship between the mortality of cardio-cerebrovascular disease and particulate matters after adjustment for the long-term trend of death,weather conditions, "days of the week" and other confounding factors. RESULTS: In single-pollutant model, the short-term effects of particulate matter on cardio-cerebrovascular mortality was strongest in lagged 2 days in Ningbo city, and an increase of 10 µg/m(3) in moving average concentrations (lagged 2-3 days and lagged 2-4 days) of PM2.5 and PM10 could increase the cardio-cerebrovascular mortality by 0.55% (0.23%-0.87%) and 0.53% (0.28%-0.78%), respectively. In multi-pollutant models, PM10 did remain robust after being adjusted for PM2.5 with 0.58% (0.09%-1.07%) increase in cardio-cerebrovascular mortality. The effect of PM2.5 had no statistical significantce after being adjusted for other co-pollutants. CONCLUSION: These findings suggested that the concentrations of ambient particulate matters were associated with an increased risk of daily cardio-cerebrovascular mortality in Ningbo city.

[The antifertility effect of gossypol plus testosterone and estrogen].

In order to reduce the side effect of gossypol, gossypol was used in combination with steroid hormones so that the dose of both drugs can be reduced. Silastic capsules containing testosterone (T) + estradiol (E) were implanted under the skin male wister rats for 8 weeks. After removing the implants, testosterone was given orally at the dose of 15 mg.kg-1 which is only 50% of the usual antifertility dose. Mating tests showed that the male rats became infertile. Microscopic examination of the heart, liver and kidneys showed no pathologic changes. The treated rats gained body weight as well as the controls. The fertility of the treated rats recovered four to five weeks after treatment. Thus, gossypol in combination with testosterone and estrogen exhibited a low degree of side effect and high antifertility activity.

Feedback loops blockade potentiates apoptosis induction and antitumor activity of a novel AKT inhibitor DC120 in human liver cancer.

The serine/threonine kinase AKT is generally accepted as a promising anticancer therapeutic target. However, the relief of feedback inhibition and enhancement of other survival pathways often attenuate the anticancer effects of AKT inhibitors. These compensatory mechanisms are very complicated and remain poorly understood. In the present study, we found a novel 2-pyrimidyl-5-amidothiazole compound, DC120, as an ATP competitive AKT kinase inhibitor that suppressed proliferation and induced apoptosis in liver cancer cells both in vitro and in vivo. DC120 blocked the phosphorylation of downstream molecules in the AKT signal pathway in dose- and time-dependent manners both in vitro and in vivo. However, unexpectedly, DC120 activated mammalian target of rapamycin complex 1 (mTORC1) pathway that was suggested by increased phosphorylation of 70KD ribosomal protein S6 kinase (P70S6K) and eukaryotic translation initiation factor 4E binding protein 1 (4E-BP1). The activated mTORC1 signal was because of increase of intracellular Ca(2+) via Ca(2+)/calmodulin (CaM)/ signaling to human vacuolar protein sorting 34 (hVps34) upon AKT inhibition. Meanwhile, DC120 attenuated the inhibitory effect of AKT on CRAF by decreasing phosphorylation of CRAF at Ser259 and thus activated the mitogen-activated protein kinase (MAPK) pathway. The activation of the mTORC1 and MAPK pathways by DC120 was not mutually dependent, and the combination of DC120 with mTORC1 inhibitor and/or MEK inhibitor induced significant apoptosis and growth inhibition both in vitro and in vivo. Taken together, the combination of AKT, mTORC1 and/or MEK inhibitors would be a promising therapeutic strategy for liver cancer treatment.

Phytochemicals potentiate interleukin-2 generated lymphokine-activated killer cell cytotoxicity against murine renal cell carcinoma.

The success of adoptive immunotherapy using recombinant interleukin-2 (rIL-2) and lymphokine-activated killer (LAK) cells in several cancers has been hampered by severe toxicity associated with high doses of rIL-2. Methods that reduce the dosage of rIL-2 without loss of clinical efficacy are needed. In this study we determined the in vitro effect of a phytochemical immune modulator, Astragalus membranaceus (AM), and two fractions isolated by high-performance liquid chromatography on the cytotoxicity of rIL-2-generated LAK cells against a murine renal cell carcinoma. Our results indicated a 10-fold potentiation of rIL-2-generated LAK cell cytotoxicity manifested by tumor cell lysis of 88% in the group with 100 U/ml of rIL-2 plus AM versus 86% in the group with 1,000 U/ml of rIL-2 alone. Potentiation was obtained with the purified fractions as well. A significantly reduced number of LAK cells was required to achieve the tumor cytotoxicity after LAK cell generation with rIL-2 plus the phytochemicals as compared with rIL-2 alone. Our data indicate that AM is an effective immune modulator, capable of potentiating in vitro the antitumor activity of rIL-2-generated LAK cells.