Lipopolysaccharide (LPS)-induced inflammation in RAW264.7 cells is inhibited by microRNA-494-3p via targeting lipoprotein-associated phospholipase A2.

BACKGROUND: Gram-negative bacterial lipopolysaccharide (LPS) is a major component of inflammation and plays a key role in the pathogenesis of sepsis. According to our previous study, the expression of lipoprotein-associated phospholipase A2 (Lp-PLA2) is significantly upregulated in septic patients and is positively correlated with the severity of this disease. Herein, we investigated the potential roles of Lp-PLA2-targeting microRNAs (miRNAs) in LPS-induced inflammation in murine mononuclear macrophages (RAW264.7 cells). METHODS: In LPS-stimulated RAW264.7 cells, Lp-PLA2 was confirmed to be expressed during the inflammatory response. The function of microRNA-494-3p (miR-494-3p) in the LPS-induced inflammatory response of RAW264.7 cells was determined by the transfection of a miR-494-3p mimic or inhibitor in vitro. RESULTS: Compared to the control, LPS induced a significant increase in the Lp-PLA2 level, which was accompanied by the release of inflammatory mediators. The bioinformatics and qRT‒PCR results indicated that the miR-494-3p level was associated with Lp-PLA2 expression in the LPS-induced inflammatory response of RAW264.7 cells. Dual-luciferase reporter assay results confirmed that the 3'-UTR of Lp-PLA2 was a functional target of microRNA-494-3p. During the LPS-induced inflammatory response of RAW264.7 cells, targeting Lp-PLA2 and transfecting miR-494-3p mimics significantly upregulated the expression of miR-494-3p, leading to a reduction in the release of inflammatory factors and conferring a protective effect on LPS-stimulated RAW264.7 cells. CONCLUSION: By targeting Lp-PLA2, miR-494-3p suppresses Lp-PLA2 secretion, thereby alleviating LPS-induced inflammation, which indicates that miR-494-3p may be a potential target for sepsis treatment.

The predictive value of CHA2DS2-VASc score on the prognosis of patients with atrial fibrillation based on a prospective cohort study.

Background: Atrial fibrillation (AF) is the most prevalent cardiac arrhythmia encountered in clinical practice, and it is associated with an increased risk of mortality, stroke, and peripheral embolism. The risk of stroke in AF is heterogeneous and dependent on underlying clinical conditions included in current risk stratification schemes. Recently, the CHA2DS2-VASc score has been incorporated into guidelines to encompass common stroke risk factors observed in routine clinical practice. The aim of this study was to study the predictive value of CHA2DS2-VASc score on the prognosis of patients with AF to determine the correlation of major complications including cerebral infarction and intracranial hemorrhage in patients with AF with oral anticoagulant and antiplatelet aggregation drugs and to identify the risk factors for all-cause mortality. Methods: A prospective study was conducted on 181 patients with AF who underwent physical examinations at Hai'an Qutang Central Hospital from January 2020 to December 2020. The patient's general condition, chronic disease history, CHA2DS2-VASc [congestive heart failure, hypertension, age ≥75 years (doubled), diabetes, stroke (doubled), vascular disease, age 65 to 74 years, and sex category (female)] score, left ventricular ejection fraction (LVEF), lipid metabolism, and oral anticoagulant and antiplatelet aggregation medication during physical examination were recorded. By using telephone meetings to complete the follow-up, we tracked the patient's cerebral infarction, intracranial hemorrhage, and survival status within 2 years of follow-up, statistically analyzed the relationship between AF complications and medication, and grouped patients with AF based on the CHA2DS2-VASc score to evaluate its predictive ability for mortality outcomes in these patients. Results: The patients were divided into four groups according to the medication situation, and the incidence of cerebral infarction in the combination group was significantly lower than that in the non-medication group (0.0% vs. 19.2%; P<0.01). The incidence of intracranial hemorrhage in the combination group was significantly higher than that in the non-drug group (13.8% vs. 0.0%; P<0.01). The logistic regression model indicated that patients with a history of cerebral infarction had an increased risk of death compared to those without a history of cerebral infarction [odds ratio (OR) =7.404; 95% confidence interval (CI): 2.255-24.309]. After grouping according to the CHA2DS2-VASc score, we found that there was a significant difference in the 2-year survival rate between patients with CHA2DS2-VASc score <5 and those with a score ≥5 (P<0.01). The characteristic curve analysis of the participants showed that the CHA2DS2-VASc score had good predictive ability for all-cause mortality in patients with AF (area under the curve =0.754), with a cutoff value of 4, a sensitivity of 62.50%, a specificity of 86.06%, and a 95% CI of 0.684-0.815. Conclusions: The CHA2DS2-VASc score demonstrated high predictive value for all-cause mortality in patients with AF.

In vivo flow cytometry reveals an anti-metastatic effect of Rujifang in triple-negative breast cancer.

Breast cancer is the most common cancer, and triple-negative breast cancer (TNBC) has the highest metastasis and mortality rate among all breast cancer subtypes. Rujifang is a traditional Chinese medicine formula with many years of clinical application in breast cancer treatment. Here, we aim to investigate the effects of Rujifang on circulating tumor cell (CTC) dynamics and the tumor microenvironment in a ZsGreen/luciferase double-labeled TNBC orthotopic model. We report that the number of CTCs monitored by in vivo flow cytometry (IVFC) strongly correlates with disease progression. Rujifang treatment decreased the number of CTCs and suppressed the distant metastasis of TNBC. Moreover, immunofluorescence analysis revealed that Rujifang treatment could affect the tumor microenvironment by downregulating Kindlin-1, which has been reported to promote metastasis of TNBC. Our study provides evidence of the anti-metastatic effect of Rujifang against TNBC in an animal model using fluorescent cell lines. The results suggest the potential therapeutic value of Rujifang as an anti-metastatic drug, however, further clinical trials are needed to validate these findings in humans.

Development and validation of a predictive model for in-hospital mortality in patients with sepsis-associated liver injury.

Background: Sepsis is often accompanied by organ dysfunction and acute organ failure, among which the liver is commonly involved. Sepsis patients suffering from liver injury have a greater risk of mortality than patients suffering from general sepsis. As of now, there are no tools that are specifically designed for assessing the prognosis of such patients. This study aimed to develop and validate a model to predict the risk of in-hospital mortality in patients with sepsis-associated liver injury (SALI). Methods: Data were obtained from the Medical Information Mart for Intensive Care (MIMIC)-IV database. In the analysis, all patients with SALI who met the inclusion and exclusion criteria were included. A primary outcome was in-hospital mortality, and clinical data were extracted for these patients. In a ratio of 8:2, the data were divided into training and validation groups at random. Least absolute shrinkage and selection operator (LASSO) regression was used for data dimension reduction and feature selection, and independent factors related to prognosis were identified through multi-factor logistics analysis. A nomogram was developed to visualize the model, and the performance of the model was evaluated by the area under the curve (AUC) as well as calibration and decision curve analysis (DCA) through internal verification. Results: A total of 616 and 154 patients with SALI were included in the training and validation cohorts, respectively. The LASSO regression and logistic multivariate analysis showed that nine factors were associated with in-hospital mortality in patients with SALI. Both the training and validation cohorts had higher AUCs than sequential organ failure assessment (SOFA) and simplified acute physiology score 2 (SAPS2): 0.753 (95% CI: 0.715-0.791) and 0.783 (95% CI: 0.749-0.817), respectively. Both the training and validation cohorts showed good calibration results for the prediction model. In terms of clinical practicability, DCA of the predictive model demonstrated greater net benefits than the SOFA and SAPS2 scores. Conclusions: We developed a predictive model that can effectively predict the in-hospital mortality of SALI patients, with satisfactory performance and clinical practicability. This model can assist clinicians in the early identification of high-risk patients and provide a reference for clinical treatment strategies.

MiR-702-3p inhibits the inflammatory injury in septic H9c2 cells by regulating NOD1.

BACKGROUND: Cardiac insufficiency is a common complication of sepsis and septic shock and is the most common cause of death in critically ill patients. Recent studies have found that microRNAs (miRNAs) play a potential role in sepsis as markers, but little is known about their functional effects on sepsis-induced cardiomyopathy (SIC). OBJECTIVE: This study is designed to explore the possible role and underlying mechanisms of miR-702-3p in septic cardiomyopathy. METHODS: As expected, H9c2 cells were induced with lipopolysaccharide (LPS) to construct the model of septic cardiomyopathy. The expression of miR-702-3p was detected by qRT-PCR assay and those of IL-1ß, IL-6 and TNF-α by ELISA assay. The viability, proliferation and apoptosis of LPS-treated H9c2 cells were determined by CCK-8, EdU, flow cytometry and western blot assays. Moreover, Nucleotide-binding oligomerization domain-containing protein 1 (NOD1) was predicted and confirmed as a direct target of miR-702-3p by TargetScan, miRwalk and miRDB prediction and dual-luciferase reporter gene assays. RESULTS: While LPS can weaken the viability of H9c2 cells, miR-702-3p enhances that of LPS-treated H9c2 cells by inhibit the expressions of TNF-α, IL-6, IL-1ß. We found NOD1 is a target gene of miR-702-3p, and over-expression of NOD1 restores the inhibitory effects of miR-702-3p on the LPS-treated H9c2 cells. CONCLUSION: MiR-702-3p played an important role in the pathogenesis of sepsis cardiomyopathy via targeting NOD1, suggesting that miR-702-3p may be a potential new target for the treatment of SIC.

The expression of STEAP4 in peripheral blood predicts the outcome of septic patients.

BACKGROUND: Sepsis is a systemic disease characterized by extensive inflammatory responses and impaired organ function, which are characteristics that make it easily missed and complex to treat. A large number of laboratory and clinical studies on the diagnosis and treatment of sepsis have been continuously carried out, confirming the importance of mitochondrial function during the development of sepsis. STEAP4 is an important metalloreductase in mitochondria, which is involved in the biogenesis and respiratory chain of mitochondria. The role of STEAP4 in inflammation remains controversial. Research in this field may contribute to the development of new diagnostic and treatment options for sepsis. METHODS: The expression of STEAP4 was measured in the peripheral blood of patients with severe sepsis and compared with healthy controls. Cell and mouse inflammatory models were established to detect the expression of STEAP4 and other inflammatory cytokines. RESULTS: (I) The expression of STEAP4 in the peripheral blood of patients with severe sepsis is higher than that of healthy volunteers (P<0.01), which is related to the SOFA score and transaminase. (II) STEAP4 has a certain predictive effect on the outcome of patients [area under curve (AUC) =0.696, P<0.05, 95% CI: 0.528 to 0.833]. (III) Inflammation led to increased expression of STEAP4 gene in RAW264.7 cells and mouse liver tissue. CONCLUSIONS: The expression of STEAP4 is elevated in the early stage of sepsis and the degree of its elevation can be used to predict the clinical outcome of sepsis patients.

Analysis of the epidemiological and clinical characteristics of 65 patients with scrub typhus on the east coast of China.

BACKGROUND: The present study set out to investigate the epidemiological and clinical characteristics of 65 patients with tsutsugamushi disease. METHODS: The clinical data of 65 patients with tsutsugamushi disease, who were admitted to the Affiliated Hospital of Nantong University were retrospectively analyzed. The clinical symptoms, laboratory examination results, clinical treatment plans, treatments, and outcomes of the patients were analyzed. RESULTS: The 65 patients with tsutsugamushi disease, included 40 males (61.54%) and 25 females (38.46%). The patients were aged from 1 year and 7 months to 88 years old, and the peak age was 60-70 years old. Geographically, the patients were concentrated in Rugao and Tongzhou District. Infections were most common between October and December (categorized as "autumn type"), and peaked in November. Farmers had the highest infection rate of any occupation (66%). All patients had the symptom of fever, with the body temperature of 60 (92.31%) patients exceeding 38.5 °C, while 58 (89%) and 51 (78%) patients had characteristic eschar and skin rash, respectively. There were 56 (86.15%) patients with varying degrees of liver damage, 8 (12.31%) cases of elevated D-dimer, 3 (4.62%) cases of myocardial injury, 38 (58.46%) cases of superficial lymph node enlargement, 8 (12.31%) cases of splenomegaly, and 2 cases (3.08%) of bulbar conjunctival congestion. Of the 65 patients enrolled, the overall misdiagnosis rate of first medical contact was 64.62. CONCLUSIONS: Tsutsugamushi disease, infection has obvious seasonality and a susceptible population, especially among farmers and the elderly.

In vitro bacteriostatic effects of Polymyxin B combined with Propofol medium and long chain fat emulsion injection against Escherichia coli.

BACKGROUND: Polymyxins is a class of cyclic polypeptide antibiotics with strong antibacterial activity against Gram-negative bacteria. However, bacteria become resistant to Polymyxins. Thus, Polymyxin B (PMB) in combination with other antimicrobials may be a better choice in clinic. This study aimed to evaluate the synergistic bacteriostatic effect of PMB combined with Propofol medium and long chain fat emulsion injection against Escherichia coli in vitro. METHODS: The Minimal Inhibitory Concentration of Polymyxin B combined with Propofol medium and long chain fat emulsion injection and two drugs used alone against Escherichia coli were detected with the Kirby-Bauer disk diffusion (K-B) method, and the diameter of the inhibition zone was calculated to evaluate bacteriostatic effects. RESULTS: Different concentrations of PMB all had obvious bacteriostatic effects on Escherichia coli, while different concentrations of Propofol medium and long chain fat emulsion injection had no bacteriostatic effects on Escherichia coli. The bacteriostatic effect of the combination of PMB with Propofol medium and long chain fat emulsion injection against Escherichia coli was synergistic, and no effects of uncorrelated and antagonism were observed in this combination. CONCLUSIONS: PMB combined with Propofol medium and long chain fat emulsion injection can improve the bacteriostatic effect for Escherichia coli in vitro.

G-rich sequence factor 1 serves as a prognostic biomarker in septic patients.

BACKGROUND: Sepsis is a condition of organ dysfunction caused by infection, and is unavoidably related to costs and mortality; however, no biomarker has yet been identified to clearly predict the prognosis of septic patients. In this study, we aimed to explore the role of guanine-rich sequence factor 1 (GRSF1) in evaluating the severity and prognosis of sepsis. METHODS: The expression of GRSF1 in peripheral blood was measured and analyzed in 42 septic participants and 32 healthy controls respectively by using quantitative reverse transcription polymerase chain reaction (RT-qPCR). Clinical data were assessed by correlation analysis. In addition, GRSF1 expression was investigated in cecal ligation and puncture (CLP) induced mice septic models by RT-qPCR and western blot (WB). RESULTS: The expression of GRSF1 expression in septic patients in the first day of electronic intensive care unit (eICU) administration was significantly lower in comparison with HC. Further analysis showed GRSF1 expression was strongly related to the Acute Physiologic Assessment and Chronic Health Evaluation II (APACHE II) score and Sequential Organ Failure Assessment (SOFA) score. Low expression of GRSF1 predicted high mortality within 24 hours in septic patients and in CLP-induced mice. CONCLUSIONS: Decreased expression of GRSF1 was significantly correlated with high mortality in septic patients, and also in experimental septic mice. The GRSF1 protein may be a potential prognostic biomarker in sepsis.

Research progress of circRNA as a biomarker of sepsis: a narrative review.

OBJECTIVE: Explore the possibility of circRNAs as markers of sepsis. BACKGROUND: Sepsis is an abnormal immune response of our body to infection that can lead to organ failure and death. Although the research on sepsis has been extensive in the past few years, sepsis-associated morbidity and mortality are still increasing. Early diagnosis and early treatment are important for patients with sepsis. Although many markers, including procalcitonin and C-reactive protein, have been proposed as diagnostic indicators of sepsis, there are still challenges in the early diagnosis and treatment of sepsis due to the lack of sensitivity and specificity of these substances. Recently, a large number of studies have found that circular RNAs (circRNAs) participate in a variety of biological functions, such as immune response, regulating the expression of miRNAs, and they are closely related to the occurrence and development of many diseases, including sepsis. However, the clear mechanism of the role of circRNAs has not been fully elucidated. An increasing number of studies have confirmed that circRNAs have potential in the diagnosis and treatment of sepsis. By studying the regulatory mechanism of circRNAs in sepsis, we can search for new molecular intervention targets for the treatment of sepsis, which is conducive to the development of new molecular therapeutic drugs for sepsis. METHODS: In the present study, we summarize and analyze the role of circRNAs in the pathogenesis of sepsis and discuss the possibility of circRNA as a biomarker for the diagnosis of sepsis. CONCLUSIONS: The biological characteristics of circRNAs and their role in the occurrence and development of sepsis make them possible markers of sepsis.

The effect of HES130/0.4 sodium chloride solution on kidney function following early fluid resuscitation in shock patients.

BACKGROUND: Doctors often use a small dose of hydroxyethyl starch (HES) 130/0.4 sodium chloride solution in the emergency room; however, its effect on kidney function remains controversial. This study aimed to evaluate the effect of a small dose of HES130/0.4 sodium chloride solution on kidney function in shock patients during early fluid resuscitation. METHODS: This cohort study retrospectively analyzed the data of 129 shock patients requiring fluid resuscitation who had been admitted to the Emergency Department of the Affiliated Hospital of Nantong University from January 2019 to December 2020. Patients were divided into the observation group (n=40) and control group (n=89) according to the type of fluid resuscitation. In relation to the fluid resuscitation treatment, the observation group was treated with crystalloid solution, while the control group was treated with crystalloid and HES130/0.4 sodium chloride solution. To further explore the effect of a small dose of HES130/0.4 sodium chloride solution, the patients were further divided into the following 4 groups based on the specific fluid administered: (I) the HES(+), lactated Ringer's (LR)(+) group (n=85); (II) the HES(+), LR(-) group (n=4); (III) the HES(-), LR(+) group (n=31); and (IV) the HES(-), LR(-) group (n=9). The outcomes were in-hospital mortality and changes in creatinine (CR) level after fluid resuscitation. RESULTS: There were no significant differences in the in-hospital mortality rates between the observation and control groups (P=0.343). The CR levels of patients in the control and HES(+), LR(+) groups were reduced after fluid resuscitation (P=0.034; P=0.028). There was no significant change in patients' CR levels in the HES(+), LR(-) group after fluid resuscitation (P=0.999). CONCLUSIONS: Administering a small dose of HES 130/0.4 sodium chloride in patients with shock does not appear to affect kidney function and in-hospital mortality; however, these findings should be considered exploratory, and further studies should be conducted to confirm these results.

Diabetes health management strategy based on internet plus graded diagnosis and treatment strategy.

BACKGROUND: Type 2 diabetes (T2D) is a widespread chronic disease with high rates of morbidity and mortality worldwide. Managing risk factors can effectively prevent acute and chronic complications, improve quality of life, and reduce mortality. Therefore, implementation of a diabetes health management strategy is urgently needed. Emerging medical technologies have strengthened communication between patients and clinicians. The establishment and improvement of the graded diagnosis and treatment system has promoted the prevention, treatment, and management of diabetes. METHODS: A total of 300 patients diagnosed with T2D in the Health Management Center at the Affiliated Hospital of Nantong University were randomly divided into two groups: an internet plus graded diagnosis and treatment strategy group, and a control group. After 6 months, the physiological parameters, management indices, and complications were compared between the groups. RESULTS: Physiological indicators, such as body mass index (BMI), waist circumference, triglycerides, low-density lipoprotein, systolic and diastolic blood pressure, and blood glucose were significantly alleviated in the Internet plus graded diagnosis and treatment strategy group. Management indicators (such as blood glucose monitoring compliance rate, diet control compliance rate, and exercise compliance rate) also improved substantially. The incidence of hypoglycemia was notably increased compared to the control group. CONCLUSIONS: The new health management strategy for diabetes can improve lifestyle, ameliorate physiological indicators, reduce the complication rate, and form a virtuous cycle. This provides a positive impact on the entire life-cycle health management of diabetes, and is worthy of further promotion.

The role of lipoprotein-associated phospholipase A2 in acute kidney injury of septic mice.

BACKGROUND: This experiment aimed to investigate the role and mechanism of lipoprotein-associated phospholipase A2 (Lp-PLA2) in kidney injury in septic mice induced by cecal ligation and perforation (CLP). METHODS: Male BALB/c mice were randomly divided into two groups: sham-operation group (Sham group) and septic group (CLP group). The septic model was simulated by cecal ligation and puncture method, but only cecal ligation was used for the sham operation group. The whole serum and renal tissue samples of the mice were collected 24 hours after modeling in both groups. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of renal tissue, the renal injury score was recorded, and the creatinine (Cr) and blood urea nitrogen (BUN) levels were detected by automatic biochemical analyzer, while the serum Lp-PLA2 level was detected by enzyme-linked immunosorbent assay (ELISA). The 7-day survival rate and the survival curve of the two groups were statistically analyzed. RESULTS: Compared with the Sham group, the pathological score of renal injury in the CLP Group was higher, the level of Lp-PLA2 in serum was significantly increased (all P<0.01), and the expression of Lp-PLA2 in renal tissue was significantly elevated (all P<0.01). Furthermore, the 7-day survival rate of the Sham group was 90%, while that of CLP group was 25%. CONCLUSIONS: The expression level of Lp-PLA2 in blood and kidney tissue of septic mice was increased and correlated with prognosis. However, the predictive value of Lp-PLA2 for prognosis in septic mice needs further study.

A clinical study of preoperative carbohydrate administration to improve insulin resistance in patients with multiple injuries.

BACKGROUND: The purpose of this study was to investigate the tolerance and safety of carbohydrate administration to patients with multiple injuries prior to surgery, and to analyze the effects of carbohydrate intake on their immediate insulin resistance (IR), postoperative complications, and length of hospital stay. METHODS: A total of 125 patients with mild multiple injuries who were admitted to the Emergency Surgery Department of Affiliated Hospital of Nantong University for elective surgery were randomized to administration of either placebo or carbohydrate. Finally, 82 patients (male: 39, female: 43) successfully completed the experiment and collected data. Preoperative general condition, subjective comfort, blood glucose concentration, serum insulin and insulin resistance index (IR) were studied. RESULTS: The two groups of patients matched in gender, age, body mass index (BMI) (P>0.05). Patients in CHO group treated with carbohydrates three hours before surgery compared with patients treated with preoperative specification. The thirst, hunger and anxiety of the patients in the CHO group were significantly relieved (P<0.05). Blood glucose concentration, serum insulin, and IR were much lower in the CHO group (P<0.05). CONCLUSIONS: It is a relatively safe approach that patients took carbohydrates 3 hours before surgery, and there was no statistically significant difference in the incidence of postoperative aspiration. Taking carbohydrates before surgery can not only relieve preoperative discomfort, but also reduce postoperative insulin resistance, which is helpful to avoid postoperative metabolic disorder and speed up recovery.

The correlations between the serum expression of miR-206 and the severity and prognosis of sepsis.

BACKGROUND: An accurate assessment of the severity and prognosis of sepsis, especially septic shock, is vital for the tailored treatment of this condition. miRNA participates in the inflammatory response and cell apoptosis and regulates inflammation-related signaling pathways. Immune disorders often accompany sepsis. Since serum miRNA expression is superior to traditional biological markers in terms of sensitivity and specificity, its role in the assessment of sepsis has increasingly been recognized. METHODS: Serum miRNAs were extracted from septic patients and healthy individuals by using the ultracentrifugation method. The differential expressions of miRNAs in the serum samples were detected by high-throughput sequencing technology. The differentially expressed miRNAs between the two groups were analyzed by bioinformatics. The quantitative polymerase chain reaction real-time polymerase chain reaction (qRT-PCR) was used to amplify the sample size to verify the results and to screen the highly-expressed miR206 in septic patients. Subsequently, serum samples were collected from 63 septic patients, and 30 patients with septic shock and qRT-PCR were performed to analyze the expression of miR-206. These 93 patients were divided into the miR-206 low-expression group and miR-206 high-expression group according to miR206 expression level. The potential correlations between the miR-206 expression and the clinical data were analyzed by using SPSS 25.0. RESULTS: Serum miRNA expression significantly differed between septic patients and healthy individuals. High-throughput sequencing results showed that, compared with those in healthy individuals, 29 miRNA molecules were down-regulated, and 25 molecules were up-regulated in the serum samples of septic patients. qRT-PCR identified the significantly up-regulated miR-206 in septic patients. qRT-PCR also showed significantly higher miR-206 expression levels in patients with septic shock than in septic patients. Furthermore, we observed a significantly longer prothrombin time and activated partial thromboplastin time, and significantly higher SOFA score, APACHE-II score, and in-hospital mortality rate. miR-206 was positively correlated with SOFA sore and APACHE-II score. CONCLUSIONS: Serum miR-206 expression is positively correlated with the severity and prognosis of sepsis. Thus, it may be a potential biomarker for assessing the severity and prognosis of sepsis, although the specific mechanism warrants further investigations.

Preparation of doxorubicin-loaded collagen-PAPBA nanoparticles and their anticancer efficacy in ovarian cancer.

BACKGROUND: The aims of this study were to prepare the collagen-poly (3-acrylamidophenylboronic acid) nanoparticles (collagen-PAPBA NPs) encapsulating doxorubicin (DOX) and research their anticancer efficacy in ovarian cancer. METHODS: Collagen-PAPBA NPs were prepared, and their morphology and stability morphology were observed by transmission electron microscopy (TEM) and dynamic light scattering system (DLS). Preparation of doxorubicin-loaded Collagen-PAPBA NPs (DOX-loaded NPs) were then prepared, and the drug-loading content, encapsulation efficiency, and in vitro drug-release profiles were calculated. The morphology of DOX-loaded NPs was also observed by DLS, in vitro cytotoxicity to A2780 cells was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, in vitro antitumor activity on A2780 cells was observed by immunofluorescence, and in vivo antitumor activity was assessed using an experimental BALB/c mice tumor model. RESULTS: DOX-encapsulating collagen-PAPBA NPs were successfully prepared with mediation by biomolecule. The average hydrodynamic diameter of collagen-PAPBA NPs as measured by DLS was about 79 nm, with a homogeneous distribution of size. TEM revealed that nanoparticles were well-dispersed, spherical, and a roughly uniform 75 nm in size. Collagen-PAPBA NPs were quite stable in a wide range of pH and temperature conditions and associated with the concentration of glucose. DLS revealed that the average hydrodynamic diameter of DOX-loaded NPs was about 81.3 nm, with homogeneous distribution of size. TEM revealed that drug-loaded nanoparticles were spherical, well-dispersed, and gad a roughly uniform size of 79 nm. The proportion of DOX loaded into the nanoparticles was 10%, while the encapsulating efficiency was 97%. The result of the releasing test showed that the drug-loaded nanoparticles, as carriers for DOX, had a good sustained-release effect. The cell toxicity experiment showed that the blank NPs had no cytotoxicity to A2780 cells, and that the drug-loaded NPS had good a sustained-release function. They may thus have potential toxic-reducing side effects. CONCLUSIONS: Under the same doses, the drug-loaded NP had a superior inhibitory effect to free DOX on the growth of human ovarian cancer.

microRNA-132 inhibits the proliferation, migration, and invasion of ovarian cancer cells by regulating CT10 oncogenic gene homolog II-related signaling pathways.

BACKGROUND: Despite a large amount of evidence showing the involvement of microRNA-132 (miR-132) in the occurrence and prognosis of many different types of cancer, the role of miR-132 in ovarian cancer and its potential molecular mechanism have yet to be fully explained. METHOD: We studied the biological function and molecular mechanism of miR-132 in ovarian cancer cell lines and clinical tissue samples using quantitative reverse transcription polymerase chain reaction (qRT-PCR), Western blot, Luciferase reporter assay, CCK8 test, colony formation test, and scratch and Transwell assays. RESULTS: The expression level of miR-132 was significantly reduced in ovarian cancer cell lines and clinical tissue samples. When the level of miR-132 was increased, the proliferation, colony-forming, migration, and invasion abilities of ovarian cancer cells were significantly inhibited. We found that miR-132 inhibits the expression of transcription factor CT10 Oncogenic Gene Homologue II (CRKII) through specific targeting of mRNA 3'-UTR. We also observed a significant increase in CRKII expression in ovarian cancer. Notably, CRKII expression was negatively correlated with miR-132 expression in clinical ovarian cancer tissue. Down-regulation of CRKII had a similar inhibitory effect on miR-132 overexpression in ovarian cancer cells, while excessive expression of CRKII reversed the inhibitory effect mediated by the excessive expression of miR-132. CONCLUSIONS: miR-132 inhibits the proliferation, invasion, and migration abilities of ovarian cancer cells through targeting CRKII.

Elevated serum levels of lipoprotein­associated phospholipase A2 predict mortality rates in patients with sepsis.

Sepsis remains one of the leading contributors to mortality rates in the intensive care unit (ICU) and emergency intensive care unit (EICU). Therefore, any treatments against the agents which produce sepsis in a medical emergency, are welcome. Elevated serum levels of lipoprotein­associated phospholipase A2 (Lp­PLA2) have been reported in a small cohort of patients with inflammation. The present study evaluated serum levels of Lp­PLA2 in patients with sepsis and investigated the role of Lp­PLA2 in sepsis. The investigation involved the selection of 151 patients with sepsis admitted to the emergency department of the Affiliated Hospital of Nantong University (Nantong, China) and 30 healthy controls. All patients (39 with sepsis, 55 with severe sepsis and 57 with septic shock) were examined on admission to the EICU. A complete blood count was performed, and serum levels of Lp­PLA2, C­reactive protein, procalcitonin, and interleukin 6, sequential organ failure (SOFA) scores and Acute Physiology and Chronic Health Evaluation II (APACHE II) scores were determined on hospital admission. The EICU and overall mortality rates were evaluated at baseline. The present study also assessed various laboratory parameters, clinical data and inflammatory cytokines. The patient follow up duration was 90 days. The data suggested that the serum levels of Lp­PLA2 on admission to the EICU in patients with sepsis were elevated, compared with those in healthy controls. The concentrations of Lp­PLA2 were correlated with the severity of disease, and were significantly associated with experimental markers of inflammation and established prognostic scores. In the total cohort, persistently elevated levels of Lp­PLA2 on admission for EICU treatment was a predictor of poor prognosis, and provided superior diagnostic use, compared with the prognostic scoring systems, including SOFA or APACHE II scores. Taken together, the results suggested that Lp­PLA2, with respect to other markers of inflammation, may have a role as a prognostic marker in sepsis, and provide background evidence for further trials to evaluate the clinical and pathophysiologic roles of Lp­PLA2 in sepsis. Persistently elevated serum concentrations of Lp­PLA2 indicated an unfavorable outcome in patients with sepsis. In addition, the results indicated the potential role of Lp­PLA2 as a prognostic biomarker in patients with sepsis during the early course of EICU treatment.

Synthesis of methylprednisolone loaded ibuprofen modified dextran based nanoparticles and their application for drug delivery in acute spinal cord injury.

To improve the therapeutic efficacy of spinal cord injury (SCI), the methylprednisolone was incorporated into nanoparticles based on the ibuprofen modified dextran. The ibuprofen modified dextran was synthesized using a direct esterification linkage between the carboxylic acids of hydrophobic drug and the hydroxyl groups of the polymer backbone. The morphology of methylprednisolone loaded nanoparticles was evaluated by transmission electron microscopy (TEM) and dynamic light scattering (DLS). The therapeutic efficacy of the prepared nanoparticles on the acute SCI model rats was assessed. It is demonstrated that methylprednisolone loaded ibuprofen modified dextran based nanoparticles (MP-loaded NPs) could promote the recovery of neurological deficits, enhance growth of neurons, decrease degeneration of injuried neurons and reduce the tissue tumor necrosis factor alpha (TNF-α) levels significantly in the SCI rats. Subsequently, the study indicates that synthesis of methylprednisolone loaded ibuprofen modified dextran based nanoparticles has a great potential in the synergetic effect treatment for spinal cord injury and nanoparticles based drug delivery system will become a powerful weapon of human conquest of disease.

[Effects of Modified Ruji Recipe in Preventing Relapse and Metastasis of Breast Cancer Patients with Negative Hormone Receptor].

Objective To observe the effect of Ruji Recipe (RR) (treated by syndrome typing) in preventing the relapse and metastasis of invasive ductal breast cancer patients with negative hormone receptor (HR) after surgery and chemotherapy. Methods Using a prospective, cohort method, 136 pa- tients with stage I - III C HR negative invasive ductal breast cancer were equally assigned to the treat- ment group (treated by RR in syndrome typing way) and the control group (routine follow-ups). Disease free survival (DFS) , overall survival (OS) , relapse and metastasis were observed in the two groups. Re- sults All patients were followed-up for 15 to 57 months, with the median follow-up of 44 months. The median DFS and OS had not reached. The 1. 0, 1. 5, 2. -0, and 3. 0 years DFS were 94.1 % (64/68) , 86. 4 % (51/59), 81. 8% (45/55), and 72. 0% (36/50) in the treatment group. They were 77. 9% (53/68), 67.2% (45)67), 60. 6% (40)66), and 54. 5% (36/66) in the control group. Significant difference existed in 1. 0, 1. 5, and 2. 0 years DFS between the two groups (X² = 7.403, 6.426, 6.459; P =0. 012, 0.013, 0. 016). No statistical difference existed in 1. 0, 1. 5, 2. 0, and 3. 0 years OS between the two groups (P >0. 05). Among triple negative breast cancer (TNBC) patients, 1. 0, 1. 5, 2. 0, and 3. 0 years DFS were 97. 0% (32/33), 92. 9% (26/28), 92.6% (2527), and 84. 6% (22/26) in the treatment group, 81. 5% (2227), 66. 7% (1827), 61. 5% (16/26), and 57. 7% (15/26) in the control group. Of them, significant difference existed in 1. 5, 2. 0, and 3. 0 years DFS between the two groups (X² =5. 893, 7. 293, 4. 591 ; P = 0. 015, 0. 007, 0. 032). At the end of follow-ups, relapse and metastasis occurred in 15 patients, local recur- rence in 2 patients, single organ metastasis in 6 patients, and multiple organs metastasis in 7 patients of the treatment group. The relapse and metastasis occurred in 30 patients , local recurrence in 2 patients , single organ metastasis in 12 patients, and multiple organs metastasis in 16 patients of the treatment group. Conclusions RR ( by syndrome typing) could improve DFS and delay progression of invasive ductal breast cancer patients with negative HR in the first 2 years after surgery. It also had certain value for relapse and metastasis of TNBC patients within 2 years.