Effect of MT2A on apoptosis and proliferation in HL60 cells.

Although accumulating evidence has revealed that metallothioneins (MTs) and its family member MT2A are strongly linked to the risk of various solid tumors, researches on the occurrence and development of acute myeloid leukemia (AML) have rarely been investigated. Here, we constructed a lentiviral vector with MT2A over-expression and the interfering plasmids with MT2A expression inhibition to study the influence of MT2A on the bioactivities of HL60 cells. After cells were infected with a lentiviral vector containing the MT2A gene, both transcription and translation levels of MT2A were significantly increased in the over-expressed group in comparison with control groups. In vitro experiments, all results demonstrated that cell reproductive capacity was inhibited, but cell apoptosis rate was significantly increased. Together, the expression of apoptosis-related protein Bcl2 was remarkably reduced, while a high expression level of Bax protein was detected. Further experiments revealed that up-regulation of MT2A induced cell apoptosis and promoted G2/M phase arrest. The mechanism may be associated with down-regulated p-IκB-α and cyclinD1 expression and up-regulated IκB-α expression in the nuclear factor-kappaB (NF-κB) pathway. On the contrary, MT2A expression was down-regulated by interfering plasmids. We found that cell proliferative potential was notably increased in the interfering group compared with the negative and untreated group. What's more, MT2A may be closely related to AML cell proliferation and function via the NF-κB signal pathway.

Serum pre-albumin is prognostic for all-cause mortality in patients with community-acquired and post-operative acute kidney injury.

BACKGROUND: Acute kidney injury (AKI) is a critical clinical syndrome characterised by a rapid decrease in renal filtration, with the accumulation of products of metabolism such as creatinine and urea. In recent years, the incidence of AKI has increased not only in critically ill hospitalised patients but also in community patients. Also, the prognosis of AKI is poor and treatment is limited in these populations. The increasing incidence and poor prognosis may be the reasons why more investigators are involved in epidemiological and risk factor analysis of AKI. AIMS: To investigate the effects of these risk factors on outcomes in both community-acquired and hospitalised AKI populations to provide certain guidance for clinics and to explore the prognostic value of prealbumin on all-cause mortality in patients with community-acquired and post-operative AKI. METHODS: From 2000 to 2010, 477 patients diagnosed with AKI and treated in the Department of Nephrology, Ruijin Hospital, Shanghai Jiaotong University, were enrolled in the community-acquired AKI (CA-AKI) group and 138 patients diagnosed with AKI after an operation were enrolled in the post-operative AKI (PO-AKI) group. Data were collected at AKI onset and 1 year after discharge and analysed retrospectively. RESULTS: Compared with PO-AKI patients, more patients in CA-AKI group had chronic kidney disease, obesity and hyperlipidaemia, and fewer patients had cerebrovascular disease (CVD), anaemia, shock or arrhythmia. Risks for CA-AKI were atherosclerosis, CVD, arrhythmia, multiple organ dysfunction syndrome and usage of vasoactive agents, and risks for PO-AKI were elderly, arrhythmia and requirement of renal replacement therapy. A higher level of serum PA was associated with a better outcome in the CA-AKI group (hazard ratio 0.92, 95% confidence interval 0.85-0.996) and PO-AKI group (hazard ratio 0.91, 95% confidence interval 0.84-0.99). In the CA-AKI group, the cumulative survival rate of patients with a normal PA level (PA >20 mg/dL) was higher than that among patients with a lower PA (PA ≤20 mg/dL; 95.4% vs 88.3%, P = 0.031). Similarly, in the PO-AKI group, a normal PA level was associated with a higher survival rate (74.1% vs 47.6%, P = 0.019). CONCLUSION: Serum PA may serve as a prognostic marker for CA-AKI and PO-AKI, and further research is warranted to confirm this finding.

A Novel Method for Estimating Low-Density Lipoprotein (LDL) Levels: Total Cholesterol and Non-High-Density Lipoprotein (HDL) Can Be Used to Predict Abnormal LDL Level in an Apparently Healthy Population.

BACKGROUND We aimed to predict the abnormal LDL level by using TG, TC, HDL, and non-HDL in this study. MATERIAL AND METHODS Triglyceride (TG), total cholesterol (TC), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) data were obtained from the Laboratory Information System (LIS) for 4 years (Oct 1, 2013 to Sept 30, 2017) from among 34 270 healthy Chinese patients at Shuyang People's Hospital. TG, TC, HDL, and LDL (direct clearance method) were measured using a TBA2000FR biochemical analyzer. The non-HDL was calculated as TC minus HDL. Correlations between TG, TC, non-HDL, and LDL were analyzed using Spearman's rank correlation. Receiver operating characteristics (ROC) curve analysis was used to evaluate the predictive utility of TG, TC, and non-HDL for the abnormal LDL level (<130 mg/dL). RESULTS Both TC (r=0.870) and non-HDL (r=0.893) were significantly positively correlated with LDL. The area under curve of TC and non-HDL can be used to predict abnormal LDL levels. Optimal thresholds were 182.5 mg/Dl (4.72 mmol/L) for TC and 135.3 mg/Dl (3.50 mmol/L) for non-HDL. Based on these optimal thresholds, less than 0.5% and 0.4% of tests with elevated LDL were missed using TC and non-HDL, respectively, but the value of these missed LDL levels was not very high (<147.3 mg/dL). CONCLUSIONS If the value of non-HDL is less than 135.3 mg/Dl (3.50 mmol/L) and/or TC is less than 182.5 mg/Dl (4.72 mmol/L) for the apparently healthy populations, the LDL level will be less than 130 mg/Dl (3.36 mmol/L). TC and non-HDL can be used to predict the abnormal LDL level in apparently healthy populations.

Reference intervals of carbohydrate antigen 19-9 in the apparently healthy adult population.

BACKGROUND: To establish reference intervals of carbohydrate antigen 19-9(CA 19-9) according to the CLSI CA28-A3 guideline and to evaluate age- and gender-related variations. METHODS: Serum CA 19-9 values of 10 149 healthy subjects (from 20 years old to 60 years old) were measured from location health checkups. The relationship between CA 19-9 and age was analyzed using Spearman's approach. The reference intervals of CA19-9 were established using Q2.5 and Q97.5 , and the 90% confidence intervals of upper limits were calculated. RESULTS: The reference intervals of CA 19-9 were 1.98-25.12 U/mL for males (1.97-25.06 U/mL for 20-50 years old and 2.31-26.13 U/mL for 50-60 years old) and 2.36-29.29 U/mL for adult (20-60 years old) females. The upper limit of reference intervals for all individuals was 26.45 U/mL; the level of CA 19-9 is higher in females than males. Carbohydrate antigen (CA) 19-9 is significantly associated with aging in adult males(r = .0930, P < .0001), but not in females (P = .4734). CONCLUSIONS: Establishing reference intervals for CA19-9 and giving age-related reference intervals of CA19-9 using a big data of healthy adult, we first discovered that CA19-9 tends to increase with age in adult males but not in females.

Effect of the Nitrogen Incorporation on the Microstructure and Photoelectric Properties of N Type Nanocrystalline Silicon Thin Films.

N type silicon-rich nanocrystalline-SiN(x) ∶ H films were prepared by plasma enhanced chemical vapor deposition technique by changing NH3 flow rate. The effect of nitrogen incorporation on the microstructure and photoelectric properties of the thin films were characterized by Raman, Fourier transform infrared spectroscopy, ultraviolet-visible absorption spectra, and Hall effect measurement. The results indicated that with the increasing NH3, a phase transition from microcrystalline to amorphous silicon occured. Transmission electron microscope observation revealed that the size of silicon quantum dots could be adjusted by varying the flow rate of NH3. The microstructure order of the films reduced with increasing the flow rate of NH3, while the optical band gap increased, and the optical band tail became narrow. Meanwhile, Si­N bonds density increased and P doping was blocked. I-V testing results showed that with increasing NH3, the conductivity of films first decreased compared with nanocrystalline-Si and then increased. These behaviors reveal a competition in the mechanisms controlling the conductivity. However, with further increasing NH3, the conductivity decreased significantly due to rapid carrier recombination on the amorphous net structure.

Reference Intervals of Alpha-Fetoprotein and Carcinoembryonic Antigen in the Apparently Healthy Population.

BACKGROUND The aim of this study was to calculate 95% reference intervals and double-sided limits of serum alpha-fetoprotein (AFP) and carcinoembryonic antigen (CEA) according to the CLSI EP28-A3 guideline. MATERIAL AND METHODS Serum AFP and CEA values were measured in samples from 26 000 healthy subjects in the Shuyang area receiving general health checkups. The 95% reference intervals and upper limits were calculated by using MedCalc. RESULTS We provided continuous reference intervals from 20 years old to 90 years old for AFP and CEA. The reference intervals were: AFP, 1.31-7.89 ng/ml (males) and 1.01-7.10 ng/ml (females); CEA, 0.51-4.86 ng/ml (males) and 0.35-3.45ng/ml (females). AFP and CEA were significantly positively correlated with age in both males (r=0.196 and r=0.198) and females (r=0.121 and r=0.197). CONCLUSIONS Different races or populations and different detection systems may result in different reference intervals for AFP and CEA. Continuous reference intervals of age changes are more accurate than age groups.

ClpP affects biofilm formation of Streptococcus mutans differently in the presence of cariogenic carbohydrates through regulating gtfBC and ftf.

The abilities to form biofilms on teeth surface and to metabolize a wide range of carbohydrates are key virulence attributes of Streptococcus mutans. ClpP has been proved to play an important role in biofilm development in streptococci. Here we demonstrated that ClpP was involved in biofilm formation of S. mutans. ClpP inactivation resulted in enhanced biofilm formation or initial cell adherence in broth supplemented with sucrose, while reduced in broth supplemented with glucose or fructose. Our results also indicated that the enhanced capacities of biofilm formation and initial cell adherence were achieved through regulating the expression of a number of extracellular sucrose-metabolizing enzymes, such as glucosyltransferases (GTFB and GTFC) at early-exponential growth phase and fructosyltransferase at late-exponential growth phase in the presence of sucrose.

Laboratory reference intervals of complete blood count for apparently healthy elderly people in Shuyang, China.

BACKGROUND: Currently, there are no appropriate RIs of blood cells available for the elderly in most clinical laboratories in China. The aim of this study is to establish the RIs of complete blood cell count for apparently healthy elderly people. METHODS: Blood specimens were collected from elderly residents by standard procedures. Complete blood counts were determined by Sysmex XE-2100 analyzer. The RIs and 95% confidence intervals were calculated by the robust method recommended by CLSI C28-A3 guideline. RESULTS: RIs established for healthy elderly include: total WBC 3.63 - 10.3 x 10(9)/L for males and 3.64 - 10.3 x 10(9)/L for females; RBC 3.74 - 5.49 x 10(12)/L for males and 3.74 - 5.53 x 10(12)/L for females; Hb 109 - 167 g/L for males and 109 - 168 g/L for females; HCT 36.0 - 51.8% for males and 35.7 - 51.8% for females; MCV 86.0 - 105 fL for males and 86.2 - 106 fL for females; MCH 26.4 - 33.6 pg for males and 26.4 - 33.8 pg for females; MCHC 293 - 333 g/L for males and 291 - 335 g/L for females; RDW-SD 39.3 - 53.7 fL for males and 39.6 - 54.5 fL for females; RDW-CV 11.7 - 15.1% for males and 11.7 - 15.2% for females; PLT 122 - 355 x 10(9)/L for males and 122 - 350 x 10(9)/L for females; PCT 14.1 - 37.6 x 10(-1) mL/L for males and 13.9 - 37.9 x 10(-1) mL/L for females; MPV 11.3 - 15.5 fL for males and 11.3 - 15.5 fL for females; PDW 9.74 - 17.0% for males and 9.72 - 17.0% for females; platelet-LCR (P-LCR) 21.3 - 51.2% for males and 21.1 - 51.4% for females. CONCLUSIONS: We established scientific and reasonable RIs of blood cell analysis for the healthy elderly in our region.

Reference intervals for total bilirubin, ALT, AST and creatinine in healthy Chinese elderly.

BACKGROUND: The aim of this study was to establish the reference intervals (RIs) of total bilirubin (TBIL), alanine aminotransferase (ALT), aspartate transaminase (AST), and creatinine (CREA) for apparently healthy elderly (Han ethnicity) in Shuyang, China. MATERIAL AND METHODS: A total of 54 912 blood specimens from elderly residents age 65-104 years were collected by standard procedures in Shuyang county of Jiangsu province. TBIL, ALT, AST, and CREA for each participant were determined by automatic biochemical analyzer. Distribution and differences of TBIL, ALT, AST, and CREA were analyzed and compared between the elderly of the same age of different sexes and different ages of the same sex. RIs of TBIL, ALT, AST, and CREA were compared with the current RIs. The RIs and 95% confidence intervals were calculated using nonparametric method (2.5th-97.5th percentiles) according to the guideline of the Clinical and Laboratory Standards Institute. RESULTS: RIs established for the healthy elderly include: TBIL 7.8~30.6 µmol/L for males and 7.3~26.1 µmol/L for females; ALT 8.7~47.3 U/L for males and 8.4~45.2 U/L for females; AST 15.7~46.9 U/L for males and 15.1~46.2 U/L for females; and CREA 45.1~100.9 µmol/L for males and 38.7~85.0 µmol/L for females. Reference intervals of TBIL, ALT, AST, and CREA for male elderly were higher than those of females, and values of CREA increased with increasing age. CONCLUSIONS: We have established a panel of locally relevant RIs. It is necessary to establish scientific and reasonable RIs of TBIL, ALT, AST, and CREA for the healthy elderly in our region, which will provide a reference for clinicians and inspection officers.

Identification of differentially expressed proteins in the ovaries of menopausal women.

PURPOSE: This study investigated proteins differentially expressed in the ovaries of menopausal women in comparison to childbearing women. METHODS: Differential protein expression was screened by difference gel electrophoresis and 2-D SDS-PAGE. Four differentially expressed proteins were excised manually, identified by mass spectrometry and confirmed by immunoblot and immunohistochemistry. RESULTS: The four proteins were identified as serum amyloid P, heat shock protein 27, Glyoxalase I and Ubiquitin carboxy-terminal hydrolase. Serum amyloid P expression was significantly up-regulated in the ovaries of menopausal women by immunoblot analysis (p < 0.05), Glyoxalase I and Ubiquitin carboxy-terminal hydrolase displayed an altered expression pattern, with higher expression in the atretic follicles of menopausal women. Weak Glyoxalase I and Ubiquitin carboxy-terminal hydrolase were observed in the granulosa and theca cells of the follicles of childbearing women. Heat shock protein 27 and serum amyloid P were clearly observed in the atretic follicles of menopausal women, while their expression was restricted to the theca cells and cytoplasm of primordial follicles in the ovaries of childbearing women. All four proteins were predominantly expressed in the atretic follicles of menopausal women. CONCLUSIONS: These data suggest that the identified proteins may play a role in the regulation of follicle atresia in menopausal women, although their functions and mechanism warrant further investigation.

Contribution of ClpP to stress tolerance and virulence properties of Streptococcus mutans.

Abilities to tolerate environmental stresses and to form biofilms on teeth surface are key virulence attributes of Streptococcus mutans, the primary causative agent of human dental caries. ClpP, the chief intracellular protease of S. mutans, along with ATPases degrades altered proteins that might be toxic for bacteria, and thus plays important roles in stress response. To further understand the roles of ClpP in stress response of S. mutans, a ClpP deficient strain was constructed and used for general stress tolerance, autolysis, mutacins production, and virulence assays. Here, we demonstrated that inactivation of ClpP in S. mutans resulted in a sensitive phenotype to several environmental stresses, including acid, cold, thermal, and oxidative stresses. The ClpP deficient strain displayed slow growth rates, poor growth yields, formation of long chains, increased clumping in broth, and reduced capacity to form biofilms in presence of glucose. Mutacins production and autolysis of S. mutans were also impaired by mutation of clpP. Animals study showed that clpP mutation increased virulence of S. mutans but not significant. However, enhanced abilities to survive lethal acid and to form biofilm in sucrose were observed in ClpP deficient strain. Our findings revealed a broad impact of ClpP on several virulence properties of S. mutans and highlighted the relevance of ClpP proteolysis with progression of diseases caused by S. mutans.

A shared mechanism for TNP-ATP recognition by members of the P2X receptor family.

P2X receptors (P2X1-7) are non-selective cation channels involved in many physiological activities such as synaptic transmission, immunological modulation, and cardiovascular function. These receptors share a conserved mechanism to sense extracellular ATP. TNP-ATP is an ATP derivative acting as a nonselective competitive P2X antagonist. Understanding how it occupies the orthosteric site in the absence of agonism may help reveal the key allostery during P2X gating. However, TNP-ATP/P2X complexes (TNP-ATP/human P2X3 (hP2X3) and TNP-ATP/chicken P2X7 (ckP2X7)) with distinct conformations and different mechanisms of action have been proposed. Whether these represent species and subtype variations or experimental differences remains unclear. Here, we show that a common mechanism of TNP-ATP recognition exists for the P2X family members by combining enhanced conformation sampling, engineered disulfide bond analysis, and covalent occupancy. In this model, the polar triphosphate moiety of TNP-ATP interacts with the orthosteric site, while its TNP-moiety is deeply embedded in the head and dorsal fin (DF) interface, creating a restrictive allostery in these two domains that results in a partly enlarged yet ion-impermeable pore. Similar results were obtained from multiple P2X subtypes of different species, including ckP2X7, hP2X3, rat P2X2 (rP2X2), and human P2X1 (hP2X1). Thus, TNP-ATP uses a common mechanism for P2X recognition and modulation by restricting the movements of the head and DF domains which are essential for P2X activation. This knowledge is applicable to the development of new P2X inhibitors.

Numerical simulation of heat transfer properties of large-sized biomass particles during pyrolysis process.

During the pyrolysis process of large particles, the conduction between particles cannot be ignored. In the present work, a numerical simulation model for the pyrolysis of biomass particles was established, which takes into account the conduction within the particles. Based on this model, the temperature distribution inside the particle during the pyrolysis process was determined and the effects of particle size, moisture content, and gas velocity on heat transfer characteristics were analyzed. The results showed that the temperatures at different positions of the particles along the inflow direction were quite different, and the maximum temperature difference inside the particles was about 146.7 K for a particle diameter of 10 mm and a velocity of 0.2 m/s. During the pyrolysis process of biomass particles, there were two peaks of Nusselt number. The increase of moisture content prolonged the pyrolysis time. The pyrolysis. time of particles with moisture content of 15 % was about 1.5 times longer than that of dry particles when the particle diameter was 10 mm. Increasing the particle size decreased the difference between the two peaks and increased the time interval between the two peaks. Increasing the gas velocity can improve the heat transfer, but the effect of too high gas velocity on improving the heat transfer is limited. The present study is of great importance for a detailed understanding of the pyrolysis process of biomass particles.

[CD5 expression is an adverse prognostic factor in diffuse large B-cell lymphoma].

OBJECTIVE: To analyze CD5 expression in diffuse large B cell lymphoma (DLBCL) and to explore its relationship with the clinicopathological characteristics. METHODS: The clinical data from 160 DLBCL patients who were treated in First Bethune Hospital of Jilin University from January 2001 to December 2010 were retrospectively analyzed. Immunohistochemical staining (SP method) for CD5, CD10, bcl-6 and MUM-1 was performed on the paraffin-embedded tissue. The relationship between CD5 expression and the clinicopathological characteristics was evaluated by Chi-square test. Survival analysis adopted Kaplan-Meier analysis and Log-rank test. RESULTS: In the patients aged 60 years or older, the incidence of CD5(+) lymphoma (12/17) was significantly higher than that of CD5(-) ones (39.9%, 57/143); two or more extranodal involvements in CD5(+) patients (11/17) were more commonly found than that of CD5(-)patients (31.5%, 45/143); DLBCL-related death in CD5(+) patients (13/17) was higher than that of CD5(-) patients (37.1%, 53/143). Survival analysis showed that the overall survival (OS) and the event-free survival (EFS) of CD5(+) patients were significantly lower than those of CD5(-) patients. In the condition of different GCB type, different therapy and low IPI (0 ∼ 2), the OS of CD5(+) DLBCL patients was significantly lower than that of CD5(-) patients, while in the condition of high IPI (3 ∼ 5), the OS of CD5(+) and CD5(-) DLBCL patient had no obvious difference. CONCLUSIONS: CD5 expression is an adverse prognostic factor in DLBCL and it has more prognostic value in the condition of low IPI (0 ∼ 2).

[Study of detection and homology analysis of the 16S rRNA methylase in 374 enterobacteriaceae clinical isolates].

OBJECTIVE: To investigate the prevalence of genes encoding 16S rRNA methylase and the spreading path of these genes in 374 clinical isolate of enterobacteriaceae. METHODS: The genes encoding 16S rRNA methylase in 374 clinical isolate of enterobacteriaceae was detected with PCR. The sequence homogeny analyses were carried out with the NCBL BLAST program and enterbacterial repetitive intergenic consensus PCR (ERIC-PCR). The conjugation experiments were used to determine the transference of 16S rRNA methylase in vitro. RESULTS: In 374 clinical isolates, methylase genes were detected in 24 strains (6.41%), including 10 armA gene positive strains and 10 rmtB gene positive strains, and 4 strains with both genes positive. Highly homogenous strains were confirmed by ERIC-PCR. 45.8% (11/24) of the conjunction experiments for these strains were positive. CONCLUSION: The aminoglycoside resistance in enterobacteriaceae was concerned related to 16S rRNA methylase. The 16S rRNA methylase transferred either by clone or by plasmids horizontal spreading.

A Scale for Predicting the Outcomes of Patients with Epilepsy: A Study of 141 Cases.

OBJECTIVE: This study aimed to identify the factors relevant for developing a scale to estimate the prognosis of patients with epilepsy. METHODS: This study followed 141 patients with newly or previously diagnosed epilepsy for between four and nine years. The patients were divided into three groups on the basis of their outcomes during the follow-up period: patients that were seizure-free without anti-epileptic drugs (AEDs) (group A, n = 48), patients with pharmacoresponsive epilepsy (group B, n = 52), and patients with pharmacoresistant epilepsy (group C, n = 41). The predictors of the prognosis of epilepsy were determined using logistic regression models and optimum subsets regression, and a scale for estimating the prognosis of epilepsy (SEPE) was developed. RESULTS: The SEPE was able to distinguish between better and worse outcomes for the three groups. A score ≤3 on the SEPE predicted that a patient would become seizure-free without the use of AEDs, with a specificity of 67% and a sensitivity of 50%. A score ≤4 on the SEPE predicted that a patient may have a positive outcome; scores in this range were assigned to 97.9% of patients that were seizure-free without the use of AEDs and 65% of patients with pharmacoresponsive epilepsy, with a specificity of 80%, a sensitivity of 81%. Scores ≥6 on the SEPE predicted a poor outcome. CONCLUSION: Of the patients with a SEPE score ≤3, some were able to become seizure-free without the use of AEDs, while for other patients, it may be possible that AED use can be discontinued. Patients with a SEPE score ≤4 have the potential to achieve long-term remission. Patients with a SEPE score ≥6 are more likely to have pharmacoresistant epilepsy.

Identification of Biomarkers for Predicting Allograft Rejection following Kidney Transplantation Based on the Weighted Gene Coexpression Network Analysis.

Kidney transplantation is the promising treatment of choice for chronic kidney disease and end-stage kidney disease and can effectively improve the quality of life and survival rates of patients. However, the allograft rejection following kidney transplantation has a negative impact on transplant success. Therefore, the present study is aimed at screening novel biomarkers for the diagnosis and treatment of allograft rejection following kidney transplantation for improving long-term transplant outcome. In the study, a total of 8 modules and 3065 genes were identified by WGCNA based on the GSE46474 and GSE15296 dataset from the Gene Expression Omnibus (GEO) database. Moreover, the results of Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed that these genes were mainly involved in the immune-related biological processes and pathways. Thus, 317 immune-related genes were selected for further analysis. Finally, 5 genes (including CD200R1, VAV2, FASLG, SH2D1B, and RAP2B) were identified as the candidate biomarkers based on the ROC and difference analysis. Furthermore, we also found that in the 5 biomarkers an interaction might exist among each other in the protein and transcription level. Taken together, our study identified CD200R1, VAV2, FASLG, SH2D1B, and RAP2B as the candidate diagnostic biomarkers, which might contribute to the prevention and treatment of allograft rejection following kidney transplantation.

The protective effect of capsaicin receptor-mediated genistein postconditioning on gastric ischemia-reperfusion injury in rats.

BACKGROUND: No published study has addressed the effect of genistein postconditioning on gastric ischemia-reperfusion (GI-R) injury in rats. AIM: To examine whether capsaicin receptor-mediated genistein postconditioning protects against gastric ischemia-reperfusion injury via the PI3K/Akt signal pathway. METHODS AND RESULTS: Chloraldurat-anesthetized rats underwent occlusion of the celiac artery for 30 min, followed by 60 min of reperfusion. Based on this animal model of gastric ischemia-reperfusion injury, genistein at doses of 100, 500 or 1,000 µg/kg was administered via peripheral vein 5 min before reperfusion. The dose of 500 µg/kg was optimal for postconditioning, at which the severity of I-R-induced gastric injury significantly decreased. Immunohistochemistry also showed that gastric mucosal cell apoptosis decreased. Capsazepine (CPZ), a specific antagonist for the capsaicin receptor, was administered (1,000 µg/kg, i.v.) just before ischemia. Capsaicin (50 mg/kg, s.c.) once a day for 4 days reversed the protective effects of genistein. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting showed increased calcitonin gene-related peptide (CGRP) expression in genistein group but not in capsazepine or capsaicin group. CGRP inhibitor CGRP8-37 also prevented the effects of genistein in decreasing gastric mucosal injury index. In addition, PI3K inhibitor LY294002 (1.5 mg/kg) reversed the protective effect of genistein. Compared with genistein group, Western blots also demonstrated decreased Akt phosphorylation in LY294002 group. CONCLUSION: Our data suggest that capsaicin receptors mediated the protective effects of genistein postconditioning. CGRP secreted by activated capsaicin-sensitive neurons played an important role in the protective effects of genistein. PI3K/Akt pathway was also involved in the protective effects of genistein.

[The protective mechanism of N-acetylcysteine against ischemia/reperfusion induced gastric injury in rats].

The present study aimed to investigate the protective mechanism of N-acetylcysteine (NAC) against gastric ischemia /reperfusion (GI/R) injury in rats. After intravenous injection (IV) of NAC (150 mg/kg) into femoral vein, the rats were subjected to 30 min of ischemia induced by clamping the celiac artery followed by 60 min of reperfusion. After the gastric mucosal damage index (GMDI) had been calculated, gastric mucosal cell in situ apoptosis was detected by TUNEL method. The protein expression of p-ERK, p-JNK and NF-kappaB, and mRNA expression of TNF-alpha and Caspase-3 in gastric mucosa were evaluated by using Western-blot or RT-PCR, respectively. The results showed that NAC not only attenuated the GI-R injury, but also decreased gastric mucosal cellular apoptosis. Furthermore, NAC increased the protein expression of p-ERK, while inhibited protein expression of p-JNK, NF-kappaB in gastric mucosa. NAC also decreased the expression of TNF-alpha mRNA and Caspase-3 mRNA in gastric mucosa. Capsazepine (CPZ) (400 mg/kg, IV) reversed the protective effect of NAC against GI/R injury in rats. These results suggest that NAC can protect rats against GI/R injury. This protective effect is possibly mediated by the up-regulation of p-ERK and down-regulation of p-JNK and NF-kappaB. In addition, vanilloid receptor subtype 1 may be involved in the protective mechanism of NAC against GI/R injury.

Prognostic Signatures Based on Thirteen Immune-Related Genes in Colorectal Cancer.

BACKGROUND: The immunosuppressive microenvironment is closely related to tumorigenesis and cancer development, including colorectal cancer (CRC). The aim of the current study was to identify new immune biomarkers for the diagnosis and treatment of CRC. MATERIALS AND METHODS: CRC data were downloaded from the Gene Expression Omnibus and The Cancer Genome Atlas databases. Sequences of immune-related genes (IRGs) were obtained from the ImmPort and InnateDB databases. Gene set enrichment analysis (GSEA) and transcription factor regulation analysis were used to explore potential mechanisms. An immune-related classifier for CRC prognosis was conducted using weighted gene co-expression network analysis (WGCNA), Cox regression analysis, and least absolute shrinkage and selection operator (LASSO) analysis. ESTIMATE and CIBERSORT algorithms were used to explore the tumor microenvironment and immune infiltration in the high-risk CRC group and the low-risk CRC group. RESULTS: By analyzing the IRGs that were significantly associated with CRC in the module, a set of 13 genes (CXCL1, F2RL1, LTB4R, GPR44, ANGPTL5, BMP5, RETNLB, MC1R, PPARGC1A, PRKDC, CEBPB, SYP, and GAB1) related to the prognosis of CRC were identified. An IRG-based prognostic signature that can be used as an independent potentially prognostic indicator was generated. The ROC curve analysis showed acceptable discrimination with AUCs of 0.68, 0.68, and 0.74 at 1-, 3-, and 5- year follow-up respectively. The predictive performance was validated in the train set. The potential mechanisms and functions of prognostic IRGs were analyzed, i.e., NOD-like receptor signaling, and transforming growth factor beta (TGFß) signaling. Besides, the stromal score and immune score were significantly different in high-risk group and low-risk group (p=4.6982e-07, p=0.0107). Besides, the proportions of resting memory CD4+ T cells was significantly higher in the high-risk groups. CONCLUSIONS: The IRG-based classifier exhibited strong predictive capacity with regard to CRC. The survival difference between the high-risk and low-risk groups was associated with tumor microenvironment and immune infiltration of CRC. Innovative biomarkers for the prediction of CRC prognosis and response to immunological therapy were identified in the present study.