[Research progress on pathogenic mechanism of sepsis-induced liver injury and treatment with traditional Chinese medicine and its active components].

Sepsis is a systemic inflammatory response syndrome caused by infection, with high morbidity and mortality. Sepsis-induced liver injury(SILI) is one of the manifestations of sepsis-induced multiple organ syndrome. At present, there is no recommended pharmacological intervention for the treatment of SILI. traditional Chinese medicine(TCM), based on the holism and dialectical treatment concept, shows the therapeutic characteristics of multi-target and multi-pathway and can comprehensively prevent and treat SILI by interfering with inflammatory factors, inflammatory signaling pathways, and anti-oxidative stress and inhibiting apoptosis. This article reviewed the experimental studies on the treatment of SILI with TCM to clarify its pathogenic mechanism and therapeutic characteristics, so as to provide more ideas and directions for the development or preparation of new drugs.

Hypermethylation of thymosin ß4 predicts a poor prognosis for patients with acute-on-chronic hepatitis B liver failure.

BACKGROUND: It has been demonstrated that thymosin ß4 (Tß4) could inflect the severity of acute-on-chronic hepatitis B liver failure (ACHBLF), but the relationship between its methylation status and the prognosis of liver failure is not clear. This study aimed to determine Tß4 promoter methylation status in patients with ACHBLF and to evaluate its prognostic value. METHODS: The study recruited 115 patients with ACHBLF, 80 with acute-on-chronic hepatitis B pre-liver failure (pre-ACHBLF), and 86 with chronic hepatitis B (CHB). In addition, there were 36 healthy controls (HCs) from the Department of Hepatology, Qilu Hospital of Shandong University. The 115 patients with ACHBLF were divided into three subgroups: 33 with early stage ACHBLF (E-ACHBLF), 42 with mid-stage ACHBLF (M-ACHBLF), and 40 with advanced stage ACHBLF (A-ACHBLF). Tß4 promoter methylation status in peripheral blood mononuclear cells (PBMCs) was measured by methylation-specific polymerase chain reaction, and mRNA was detected by quantitative real-time polymerase chain reaction. RESULTS: Methylation frequency of Tß4 was significantly higher in patients with ACHBLF than in those with pre-ACHBLF, CHB or HCs. However, expression of Tß4 mRNA showed the opposite trend. In patients with ACHBLF, Tß4 promoter methylation status correlated negatively with mRNA levels. The 3-month mortality of ACHBLF in the methylated group was significantly higher than that in the unmethylated group. Also, Tß4 promoter methylation frequency was lower in survivors than in non-survivors. When used to predict the 1-, 2-, and 3-month incidence of ACHBLF, Tß4 methylation status was better than the model for end-stage liver disease (MELD) score. The predictive value of Tß4 methylation was higher than that of MELD score for the mortality of patients with E-ACHBLF and M-ACHBLF, but not for A-ACHBLF. CONCLUSIONS: Tß4 methylation might be an important early marker for predicting disease incidence and prognosis in patients with ACHBLF.

FK228 potentiates topotecan activity against small cell lung cancer cells via induction of SLFN11.

The response rate of topotecan, as a second-line chemotherapeutic drug for small cell lung cancer, is ~20%. DNA/RNA helicase SLFN11 (schlafen family member 11), a member of the Schlafen (SLFN) family, is a crucial determinant of response to many DNA damaging agents, expression of SLFN11 tends to augment the antitumor effects of the commonly used DNA-targeting agents. In the present study we investigated how SLFN11 expression regulated the sensitivity of small cell lung cancer to topotecan. We showed that SLFN11 expression levels were positively associated with the sensitivity to topotecan in a panel of seven SCLC cell lines. Topotecan treatment induced different patterns of the DNA response network in SCLC cells: DNA damage response (DDR) was more prominently activated in SLFN11-deficient SCLC cell line H82 than in SLFN11-plentiful SCLC cell line DMS273, whereas topotecan induced significant accumulation of p-Chk1, p-RPA2 and Rad51 in H82 cells, but not in DMS273 cells. We unraveled that SLFN11 expression was highly negatively correlated to the methylation of the SLFN11 promoter. HDAC inhibitors FK228 and SAHA dose-dependently increased SLFN11 expression through suppressing DNA methylation at the SLFN11 promoter, thereby sensitizing SCLC cells to topotecan. Finally, we assessed the methylation status of the SLFN11 promoter in 27 SCLC clinical specimens, and found that most of the clinical samples (24/27) showed DNA methylation at the SLFN11 promoter. In conclusion, it is feasible to combine topotecan with FK228 to improve the response rate of topotecan in SCLC patients.

Combinations of proteasome inhibitors with obatoclax are effective for small cell lung cancer.

Proteasome inhibitors, bortezomib (BTZ), and carfilzomib (CFZ) are approved drugs for hematological malignancies, but lack anticancer activities against most solid tumors. Small cell lung cancer (SCLC) is a very aggressive neuroendocrine carcinoma of the lungs demanding effective therapy. In this study we investigated whether BTZ or CFZ combined with obatoclax (OBX), an antagonist for MCL-1 and a pan-BCL family inhibitor, could cause synergistic growth inhibition of SCLC cells. We showed that combined application of BTZ or CFZ with OBX caused synergistic growth inhibition of human SCLC cell lines (H82, H526, DMS79, H196, H1963, and H69) than single agent alone. Both BTZ-OBX and CFZ-OBX combinations displayed marked synergism on inducing apoptosis (~50% increase vs BTZ or CFZ alone). A comprehensive proteomics analysis revealed that BTZ preferentially induced the expression of MCL-1, an antiapoptotic protein, in SCLC cells. Thus, proteasome inhibitor-OBX combinations could specifically induce massive growth inhibition and apoptosis in SCLC cells. Subsequent proteome-wide profiling analysis of activated transcription factors suggested that BTZ- or CFZ-induced MCL-1 upregulation was transcriptionally driven by FOXM1. In nude mice bearing in SCLC H82 xenografts, both BTZ-OBX, and CFZ-OBX combinations exhibited remarkable antitumor activities against SCLC tumors evidenced by significant reduction of tumor size and the proliferation marker Ki-67 signals in tumor tissues as compared with single agent alone. Thus, proteasome inhibitor-OBX combinations are worth immediate assessments for SCLC in clinical settings.

[Changes of wheat seed vigor at different development stages and their response to environmental temperature]. / ä¸åŒå‘è‚²æ—¶æœŸå°éº¦ç§å­æ´»åŠ›çš„å˜åŒ–åŠå ¶å¯¹çŽ¯å¢ƒæ¸©åº¦çš„å“åº”.

Two wheat cultivars (Jimai 22 and Shannong 23) were selected to investigate the changes of seed vigor at different development stages in different years by standard germination test, and the effect of environmental temperature on it was also analyzed, which would provide a theoretical basis for the utilization of early wheat seeds and the production of high vigor seeds. The results showed that the germination ability of fresh seeds appeared around 26 d after anthesis, and then the germination rate of fresh seeds displayed an overall upward trend with the development of wheat seeds. The germination energy, germination rate and germination index of dry seeds rapidly increased from 5 d to 8 d after anthesis, and then remained relatively stable, while the vigor index increased continually and reached the peak 4-6 days before the end of ripening due to the increase of seedling dry mass. The dry seeds at different developmental stages were also studied by field-plan-ting and the seed vigor of their progenies was also determined. For Jimai 22, the results indicated that the field emergence of dry seeds at 17 d after anthesis was higher and the plant could form grains, there were no significant differences in germination rate and vigor index among different samples of progeny seeds. In addition, the changes of seed vigor at different development stages were significantly affected by the environmental temperature. In the year with the high mean daily average temperature, high mean daily maximum temperature, high mean daily minimum temperature after anthesis, and large mean daily temperature difference after anthesis, the seed development time was shorter, but the 100-grain mass and seed vigor reached the peak earlier; Otherwise, the seed development time was longer, but the accumulated temperature of ripe stage was higher, and the seed vigor was higher.

[Relationships of wheat seed vigor with enzyme activities and gene expression related to seed germination under stress conditions.] / é€†å¢ƒæ¡ä»¶ä¸‹å°éº¦ç§å­æ´»åŠ›ä¸Žç§å­èŒå‘ç›¸å ³é ¶æ´»æ€§åŠå ¶åŸºå› è¡¨è¾¾çš„å ³ç³».

This study aimed to analyze the enzyme activities and gene expression differences related to seed germination in different wheat genotypes, and to clarify the relationship between seed vigor and the related enzyme activities as well as gene expression under stress germination conditions. We measured seed vigor, total soluble sugar content, soluble protein content, α-amylase activity, cysteine protease activity and gene expression of the related enzymes of four wheat cultivars under drought, artificial aging and cold soaking stress. Results showed that drought, artificial aging and cold soaking stress affected seed vigor to some extent. Under different germination conditions, total soluble sugar content showed an increasing trend with small amplitude at first and then decreased with small amplitude and after that increased rapidly again, while soluble protein content in the four cultivars gradually decreased with germination time. The α-amylase activity of the four cultivars showed a rising trend on the whole, but that of Yunong 949 and Lunxuan 061 declined after germinating for 60 hours after cold soaking stress. Cysteine protease activity decreased at first and then increased as a whole. However, under drought stress condition, cysteine protease activity of Yunong 949, Yumai 49-198 and Lunxuan 061 increased at first and then decreased and finally increased again. The α-AMY (α-amylase gene) expression levels increased at first and then decreased as a whole under different germination conditions. The expression level of α-AMY in Lunxuan 061 after cold soaking stress was higher than that of CK, while the α-AMY expression levels in four cultivars under other stress germination conditions were lower than that of CK. The expression level of CP (cysteine protease gene) showed an increasing trend. No significant difference of CP expression level was found in Chang 4738 between artificial aging treatment and CK. The CP expression levels in the four cultivars under other stress conditions were higher than that of CK. The results demonstrated that there was no direct relationship between the enzyme activities and gene expression levels of α-amylase and cysteine protease under different germination conditions. The α-amylase activity and total soluble sugar content had an extremely significant positive correlation, but the correlation between cysteine protease activity and soluble protein content was not significant. The α-amylase activity significantly positively correlated with vigor index under standard germination condition. However, the α-amylase activity had no significant correlation with vigor index under stress conditions. After cold soaking stress, cysteine protease activity significantly positively correlated with vi-gor index during seed germination, but the correlation was not significant under standard germination, drought stress and artificial aging.

[Seed vigor evaluation based on adversity resistance index of wheat seed germination under stress conditions.] / 基于小麦种子发芽逆境抗逆指数的种子活力评价.

A total of 16 wheat cultivars were selected to detect seed vigor of different genotypes using standard germination test, seed germination test under stress conditions and field emergence test. The adversity resistance indices of seed vigor indices and field emergence percentage under different germination conditions were used as the indices to evaluate adversity resistance. Principal component analysis and cluster analysis were used for the comprehensive evaluation of seed vigor. Results showed that drought stress, artificial aging and cold soaking treatments affected seed vigor to some extent. The adversity resistance indices of the artificial aging and cold soaking tests were significantly positively correlated with the field emergence percentage, while the adversity resistance index of drought stress test had no significant correlation with the field emergence percentage. 16 wheat cultivars were classified as three groups based on the principal component analysis and cluster analysis. Yunong 949, Yumai 49-198, Luyuan 502, Zhengyumai 9987, Shimai 21, Shannong 23, and Shixin 828 belonged to high vigor seeds. Xunong 5, Yunong 982, Tangmai 8, Jimai 20, Jimai 22, Jinan 17, and Shannong 20 belonged to medium vigor seeds. The other two cultivars, Chang 4738 and Lunxuan 061, belonged to low vigor seeds.

[Effects of Rapamycin and Rapamycin-loaded Poly(lactic-co-glycolic)Acid Nanoparticles on Apoptosis and Expression of bcl-2 and p27(kip1) Proteins of Human Umbilical Arterial Vascular Smooth Muscle Cell].

OBJECTIVE: To investgate the effects of rapamycin(RPM)and RPM-loaded poly(lactic-co-glycolic)acid(PLGA)nanoparticles(NPs)on the apoptosis of human umbilical arterial vascular smooth muscle cells(HUASMCs)in vitro and expression of bcl-2 and p27(kip1) protein. METHODS: HUASMCs were cultured in vitro and divided to RPM and RPM-PLGA-NPs groups treated at 3 different concentration by 12 and 24 hours,with M231-smooth muscle growth supplements medium and null-PLGA-NPs treated groups as controlled. The apoptosis of HUASMCs was determined by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling staining and flow cytometry. The expressions of bcl-2 and p27(kip1) were detected by streptacidin/peroxidase immunohistochemical method. The effect on cellular proliferation was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromidecolorimetry. RESULTS: The proliferation of HUASMCs was inhibited by RPM and RPM-PLGA-NPs in a dose-dependent manner. DNA electrophoresis showed DNA ladder in RPM and RPM-PLGA-NPs groups and classical scalar strips in control groups. The apoptotic indexes of RPM 100 ng/ml group and RPM-PLGA-NPs 500 ng/ml group detected by flow cytometry were(45.45<2.36)% and(35.04<5.64)%,respectively,which were significantly higher than that of M231-smooth muscle growth supplements control group [(2.60<0.95)%,all P<0.01]. The apoptotic indexes of groups incubated with RPM and RPM-PLGA-NPs for 24 hours were significantly higher than those of groups which incubated for 12 hours(P<0.05,P<0.01). The positive expression indexes(PEI)of p27(kip1) and bcl-2 protein were higher in RPM and RPM-PLGA-NPs groups than that of control groups. The Spearman's rank correlation coefficient test showed that there was no significant correlation between the PEI of p27(kip1) and the apoptotic indexes in the RPM group and RPM-PLGA-NPs group(P>0.05). CONCLUSIONS: Rapamycin-loaded PLGA nanoparticles and rapamycin have similar effects in inhibiting proliferation and inducing apoptosis;meanwhile,they upregulate the expression of p27(kip1) protein without downregulating the expression of bcl-2 protein in HUASMCs in vitro. RPM-PLGA-NPs has more potent pro-apoptotic effect than equivalent dose of RPM but is not linearly correlated with the p27(kip1) expression level.

Peripheral type I interferon receptor correlated with oxidative stress in chronic hepatitis B virus infection.

Type I interferon receptor (IFNAR) has been involved in the progression of chronic hepatitis B (CHB). Oxidative stress is also associated with hepatitis B virus (HBV) infection and might contribute to the structure and function of protein synthesis including the IFNAR family. This study was aimed to determine the possible associations between oxidative stress and peripheral IFNAR expression in chronic HBV infection. Fifty-four CHB patients and 31 liver cirrhosis (LC) patients were consecutively collected, as well as 11 healthy subjects as controls. Expression levels of IFNAR1 and IFNAR2 in peripheral blood lymphocytes and monocytes were measured by flow cytometry. IFNAR1 and IFNAR2c mRNA were detected by real-time reverse transcription-polymerase chain reaction. Levels of plasma-soluble IFNAR and oxidative stress parameters, including xanthine oxidase (XOD), malondialdehyde (MDA), glutathione (GSH), glutathione S-transferase (GST), and glutathione peroxidase (GSH-Px) were detected by enzyme linked immunosorbent assay (ELISA). The frequencies of IFNAR1 and IFNAR2 in lymphocytes and monocytes were significantly increased in CHB and LC patients than in healthy controls. Expression levels of IFNAR1 and IFNAR2c mRNA and plasma-soluble IFNAR level in CHB and LC patients were upregulated compared with healthy controls. Mean fluorescence intensity (MFI) of IFNAR2 in monocytes of CHB patients was higher than that in LC patients. Levels of plasma XOD, MDA, and GST were significantly increased in CHB and LC patients compared with healthy controls. Meanwhile, GSH and GSH-Px in CHB and LC patients were decreased than that in healthy controls. Furthermore, plasma MDA, GSH, and GST levels in CHB patients were higher than that in LC patients. In CHB patients, plasma GST level was negatively correlated with MFI of IFNAR2 in lymphocytes. Our results suggested that oxidative stress play an important role in the regulation of IFNAR in chronic HBV infection.

[Responses of winter wheat tillers at different positions to low temperature stress at stem elongation stage and their freezing resistance evaluation].

Taking two winter wheat cultivars Ji' nan 17 and Shannong 8355 as test materials, this paper measured the superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) activities and the malondialdehyde (MDA) and soluble protein contents in the functional leaves and sheaths of the tillers at different positions at stem elongation stage under low temperature stress, and then, the freezing resistance of the tillers was comprehensively evaluated by the methods of principal component analysis and cluster analysis. The results showed that under low temperature stress, the SOD, POD, and CAT activities in the functional leaves and sheaths of each tiller at stem elongation stage increased, but the MDA and soluble protein contents increased or decreased to some extent. By using principal component analysis and cluster analysis, the tillers of each cultivar were grouped into three kinds of freezing resistance type. For Ji' nan 17, the main stem, tiller I, and tiller II belonged to high freezing resistance type, the tiller III, tiller IV, and tiller I p belonged to medium freezing resistance type, and the tiller II p belonged to low freezing resistance type. For Shannong 8355, the main stem, tiller I, tiller II, and tiller III belonged to high freezing resistance type, the tiller IV and tiller I p belonged to medium freezing resistance type, and the tiller II p belonged to low freezing resistance type. It was concluded that the freezing resistance of the winter wheat tillers at different positions at stem elongation stage differed, with the lower position tillers being more resistant than the higher position tillers.

[Freeze resistance analysis of different wheat cultivars based on the relationships between physiological indices and grain yield].

A pot experiment with twenty wheat cultivars was conducted to investigate the superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities and the MDA, soluble protein and soluble sugar contents of functional leaves as well as the grain yield, 1000-grain weight, and grain morphological characters under low temperature stress. Low temperature (-4 degrees C) stress at stem elongation stage resulted in the changes of grain morphology and yield characters. For most of test cultivars, their grain length-width ratio, grain roundness, and sterile spikelets increased, and their grain equivalent diameter, grain area, 1000-grain weight, and grain yield decreased. Path analysis indicated that after treated with low temperature at stem elongation stage, the SOD activity and soluble sugar content of functional leaves, especially the SOD activity (direct path coefficient -0. 578) , were the dominant factors affecting grain yield. Taking the percentage of decreased grain yield due to low temperature stress as the assessment criterion, the test twenty winter wheat cultivars were divided into three groups. Cultivars Jimai 19, Jimai 20, Liangxing 99, Shannong 1135, Shannong 8355, Taishan 23, Taishan 9818, Wennong 6, and Yannong 21 belonged to high freeze resistance group, cultivars Linmai 2, Weimai 8, Yannong 19, and Zimai 12 were of low freeze resistance group, and the other seven cultivars belonged to medium freeze resistance group. The seedling stage comprehensive assessment index (D value) had a significant negative correlation with the percentage of decreased grain yield (r = -0. 512*), suggesting that the stronger freeze resistance of wheat at seedling stage was beneficial to the higher wheat grain yield, and seedling stage was the critical period to be selected to identify the freeze resistance of wheat.