[Efficacy and safety of eltrombopag in the treatment of primary immune thrombocytopenia: real-world data from a single medical center].

Objective: This study aimed at investigating the efficacy and safety of eltrombopag in the treatment of adult primary immune thrombocytopenia (ITP) and evaluated the factors influencing its efficacy and side effects. Methods: A total of 198 patients with adult ITP who were admitted to Tianjin Medical University General Hospital between January 2018 and March 2022 were retrospectively analyzed. The efficacy of each starting dose of eltrombopag was evaluated, and adverse events were analyzed. The factors influencing efficacy were investigated, including sex, age, adult ITP type, platelet antibodies, and combined drug treatments. Results: Of the 198 patients, 70 males and 128 females with a median age of 45 years (18-88 years) were included; 130 (65.7%) had newly diagnosed adult ITP, 25 (12.6%) had persistent adult ITP, and 43 (21.7%) had chronic adult ITP. The bleeding event scores at baseline were assessed; 84.3% had scores of<4 and 15.7% had scores of ≥4. The eltrombopag response rate (initial response) at 6 weeks was 78.8% (complete response [CR]: 49.0%; CR1: 14.6%; CR2: 15.2%). The median response time to eltrombopag was 7 (7, 14) days. The initial response rates to 25, 50, and 75 mg eltrombopag were 74.1%, 85.9%, and 60.0%, respectively (P=0.031). The initial response rate to the 50 mg dose was significantly higher than that of the 25-mg and 75-mg doses. Two patients received 100 mg as the starting dose, and their initial response was 0. Regarding dose adjustment, 70.7% of the patients remained on the starting dose, 8.6% underwent dose adjustment to 50 mg, and 6.1% underwent dose adjustment to 75 mg. Another two patients underwent dose adjustment to 100 mg. After dose adjustment, the persistent response rates were 83.6%, 85.3%, and 85.7% for the 25-, 50-, and 75-mg doses, respectively, with no significant difference. After dose adjustment, the sustained efficacy rate for the 100-mg dose (4 patients) was 100.0%. After 6 weeks of treatment with eltrombopag, the overall bleeding score of patients with ITP decreased. The number of patients with a score of ≥4 decreased to 0, the number of patients with a score of<4 decreased, and there was no significant change in the number of patients with a score of 1-2. The most common adverse event associated with eltrombopag was impaired liver function (7.7%). No thrombosis events or other adverse events were observed. ITP type and number of megakaryocytes significantly affected the initial response to eltrombopag. The initial response rates to eltrombopag for newly diagnosed adult ITP, persistent adult ITP, and chronic adult ITP were 85.3%, 56.0%, and 76.2%, respectively (P=0.003). For megakaryocytes, the initial response rates were 61.8%, 87.1%, and 84.3% (P=0.009) for the decreased, normal, and increased megakaryocyte groups, respectively. Conclusion: Eltrombopag, as a second-line or higher treatment for adult ITP, has a rapid onset of action and good safety. The initial response rate is significantly higher with a dose of 50 mg than with a dose of 25 mg. Patients with newly diagnosed ITP and those with normal or increased megakaryocyte numbers have a higher initial response rate to eltrombopag.

[Evaluation of the efficacy and safety of intravenous infusion of ferric derisomaltose in the treatment of iron deficiency anemia: a single-center retrospective analysis].

Objective: To investigate the clinical efficacy and safety of ferric derisomaltose injection versus iron sucrose injection in the treatment of iron deficiency anemia (IDA) . Methods: A total of 120 patients with iron deficiency anemia admitted from June 2021 to March 2023 were given intravenous iron supplementation with ferric derisomaltose to assess the efficacy and safety of hemoglobin (HGB) elevation before and after treatment. Simultaneously, the clinical effects of iron supplementation with iron sucrose were compared to those of inpatient patients during the same period. Results: Baseline values were comparable in both groups. Within 12 weeks of treatment, the elevated HGB level in the ferric derisomaltose group was higher than that of the iron sucrose group, with a statistical difference at all time points, and the proportion of HGB increased over 20 g/L in the patients treated for 4 weeks was higher (98.7%, 75.9% ). During the treatment with ferric derisomaltose and iron sucrose, the proportion of mild adverse reactions in the ferric derisomaltose group was slightly lower than that of the iron sucrose group, and neither group experienced any serious adverse reactions. The patients responded well to the infusion treatment, with no reports of pain or pigmentation at the injection site. Conclusion: The treatment of IDA patients with ferric derisomaltose has a satisfactory curative effect, with the advantages of rapidity, accuracy, and safety. Therefore, it is worthy of widespread clinical use.

[Research progress on the effects of proanthochanidins in reshaping microbiota and suppressing inflammation].

Proanthocyanidins (PCs) are a class of polyphenols that are composed of flavanate monomers and their polymers, which have antibacterial and anti-inflammatory properties with very few side effects. This article reviews the mechanism by which PCs differentially regulate microbiota, reshape microflora diversity and play a role in suppressing inflammation, providing a reference for the basic research of PCs in improving female vaginal health, and is expected to provide a new idea and breakthrough for the combined use of PCs with other antibacterial drugs in the treatment of vaginitis.

[Role and clinical significance of MUC4 gene mutations in thrombotic events in patients with classic paroxysmal nocturnal hemoglobinuria].

Objective: This study aimed to investigate the role and clinical significance of MUC4 gene mutations in thrombotic events in patients with classic paroxysmal nocturnal hemoglobinuria (PNH) patients. Methods: A retrospective analysis was conducted on the clinical data and gene sequencing results of 45 patients with classic PNH admitted to the Department of Hematology, Tianjin Medical University General Hospital, from June 2018 to February 2022. MUC4 gene mutations in patients with classic PNH were summarized, and the risk factors for thrombotic events in these patients were analyzed. Additionally, the effects of MUC4 gene mutations on the cumulative incidence and survival of thrombotic events in patients with classic PNH were determined. Results: The detection rate of MUC4 gene mutations in patients with classic PNH who experienced thrombotic events (thrombotic group) was 68.8% (11/16), which was significantly higher than that in the non-thrombotic group [10.3% (3/29) ] (P<0.001). All mutations occurred in exon 2. MUC4 mutation (OR=20.815, P=0.010) was identified as an independent risk factor for thrombotic events in patients with classic PNH. The cumulative incidence of thrombotic events was 78.6% (11/14) in the MUC4 gene mutation group (mutation group) and 16.1% (5/31) in the non-mutation group, showing a statistically significant difference between the two groups (P<0.001). Survival analysis showed a lower overall survival (OS) rate in the thrombotic group compared with that in the non-thrombotic group [ (34.4±25.2) % vs. (62.7±19.3) % ] (P=0.045). The OS rate of patients was (41.7±29.9) % in the mutation group and (59.1±18.3) % in the non-mutation group (P=0.487) . Conclusion: MUC4 gene mutations are associated with an increased incidence of thrombotic events in classic PNH patients, highlighting their role as independent risk factors for thrombosis in this population. These mutations can be considered a novel predictive factor that aids in evaluating the risk of thrombosis in patients with classic PNH.

Sun-like light source design considering non-visual performance to improve working efficiency.

With the discovery of intrinsically photosensitive retinal ganglion cells, we have a better understanding of the non-visual effects of lighting. In this study, the optimum spectral power distribution in sunlight of different color temperatures is calculated by MATLAB software. At the same time, the ratio of the non-visual effect and visual effect (K e,α ) at different color temperatures is calculated according to the sunlight spectrum to evaluate the non-visual and visual effects of white LEDs at the corresponding color temperature. Then based on the characteristics of monochromatic LED spectra, the joint-density-of-states model is used as a mathematical model, and the optimal solution is calculated for its database. According to the calculated combination scheme, Light Tools software is used to optimize and simulate the expected light source parameters. The final color temperature is 7525 K, the color coordinate is (0.2959, 0.3255), and the color rendering index reaches 92. The high efficiency light source has not only the function of lighting, but also the effect of improving work efficiency with lower blue light hazard efficiency of radiation than normal LEDs.

[Utilizing ultra-small volume graft in auxiliary liver transplantation for portal hypertension].

Objective: To examine the clinical effect of auxiliary liver transplantation with ultra-small volume graft in the treatment of portal hypertension. Methods: Twelve cases of portal hypertension treated by auxiliary liver transplantation with small volume graft at Liver Transplantation Center,Beijing Friendship Hospital, Capital Medical University between December 2014 and March 2022 were studied retrospectively. There were 8 males and 4 females,aged 14 to 66 years. Model for end-stage liver disease scores were 1 to 15 points and Child scores were 6 to 11 points. The grafts was derived from living donors in 9 cases,from split cadaveric donors in 2 cases,from whole cadaveric liver of child in 1 case. The graft recipient body weight ratios of 3 cadaveric donor livers were 0.79% to 0.90%, and of 9 living donor livers were 0.31% to 0.55%.In these cases, ultra-small volume grafts were implanted. The survivals of patient and graft, complications, portal vein blood flow of residual liver and graft, abdominal drainage and biochemical indexes of liver function were observed. Results: All the grafts and patients survived. Complications included outflow tract torsion in 2 cases, acute rejection in 1 case, bile leakage in 1 case, and thyroid cancer at the later stage of follow-up in 1 case, all of which were cured. The torsion of outflow tract was attributed to the change of anastomotic angle after the growth of donor liver. After the improvement of anastomotic method, the complication did not recur in the later stage. There was no complication of portal hypertension. The measurement of ultrasonic portal vein blood flow velocity showed that the blood flow of residual liver decreased significantly in the early stage after operation, and maintained a very low blood flow velocity or occlusion in the long term after operation, and the blood flow of transplanted liver was stable. Conclusions: Auxiliary liver transplantation can implant ultra-small donor liver through compensation of residual liver. This method may promote the development of living donor left lobe donation and split liver transplantation. However, the auxiliary liver transplantation is complex, and it is difficult to control the complications. Therefore, this method is currently limited to centers that are skilled in living related liver transplantation and that have complete ability to monitor and deal with complications.

[The association between bisphenol A exposure and obesity/overweight in children and adolescents: dose-response Meta analysis].

Objective: To explore the relationship between bisphenol A (BPA) exposure and obesity/overweight in children and adolescents through Dose-response Meta analysis. Methods: Articles published up to September 1st 2021 were systematically searched in PubMed, Web of science, Scopus, Embase, Cochrane library, CNKI, Wanfang databases and VIP Chinese Science and Technology Journal by using "bisphenol A" "BPA" "obesity" "weight" "fat""overweight" "body mass index" "BMI" "waist circumference" (both in English and in Chinese) as keywords. Use Stata 15.1 software to calculate the pooled OR (95%CI), perform heterogeneity test, subgroup analysis, sensitivity analysis, publication bias and the exposure-response relationship fitting. Results: A total of 9 English articles were included from 1 948 articles retrieved, including 8 articles from American and 1 article from China. 15 614 children/adolescents and 3 446 obese/overweight cases were further used for Meta-analysis of dose-response relationship. Meta-analysis results showed that there was heterogeneity among the highest dose groups in different studies(I2=52.1%, P=0.033). The random effect model-analysis found that compared with those in the lowest group, the OR(95%CI) for those in the highest group of urine BPA was 1.56(1.18-1.94)for the risk of obesity/overweight in children and adolescents, but there was no linear or nonlinear dose-response relationship. Sensitivity analyses showed that the results were robust, Egger's test(P=0.263) and Begg's test(P=0.348) showed that there was no publication bias. Conclusion: Bisphenol A exposure may increase the risk of obesity/overweight epidemics in children and adolescent.

[Clinical application value of peripheral blood metagenomic next-generation sequencing test for patients with hematological diseases accompanied by fever].

Objective: To investigate the clinical application value of peripheral blood metagenomic next-generation sequencing (mNGS) test for patients with hematological diseases accompanied by fever. Methods: The blood mNGS results and clinical data of inpatients with hematological diseases accompanied by fever treated in the Hematology Department of Tianjin Medical University General Hospital in March 2020 to June 2021were retrospectively analyzed. A total of 90 patients with 98 cases of specimens were included. The pathogen distribution characteristics and mNGS test performance were analyzed. Results: The positive rate of peripheral blood mNGS was significantly higher than that of traditional examination (68.37% vs 37.76%, P<0.001) and blood culture (68.37% vs 9.18%, P<0.001) . Viral, bacterial, and fungal infections accounted for 38.81%, 14.93%, and 2.99% in patients with single-pathogen infections, respectively. Polymicrobial infections accounted for 43.28%, in which viral and bacterial coinfections were the most common type (25.37%) . There were 55 virus-positive cases (82.09%) , 30 bacteria-positive cases (44.78%) , and 14 fungus-positive cases (20.90%) . The clinical approval rate of peripheral blood mNGS was 64.63% (63/98) . The sensitivity, specificity, positive predictive value, and negative predictive value (NPV) of peripheral blood mNGS were 75.68%, 36.07%, 41.79%, and 70.97%, respectively, and the overall consistency rate with traditional examination was 51.02%. Of the 22 pulmonary infection cases with no detectable pathogens by conventional tests, the pathogens were identified by peripheral blood mNGS in 14 cases, 10 of which were clinically approved. Conclusion: The positive rate of peripheral blood mNGS was significantly higher than that of blood culture and traditional laboratory examination. Peripheral blood mNGS had a high clinical recognition rate, sensitivity, and NPV in the detection of pathogens in patients with hematological diseases accompanied by fever.

[The advances on the formation and toxic effects of haloacetaldehydes disinfection by-products in drinking water].

Haloacetaldehydes (HALs), as emerging disinfection by-products in drinking water, are the third largest group by weight of identified disinfection by-products (DBPs) in drinking water. The formation of HALs is associated with the level of natural organic matter and halide in the source water, the treatment process of drinking water and the type of disinfectant. Recent studies have shown that HALs are more cytotoxic and genotoxic than regulated trihalomethanes and halo-acetic acids in drinking water. Currently, only a few countries and regions have set limit values for trichloroacetaldehyde with high detection rate in drinking water. However, there is growing evidence that unregulated HALs have a higher potential risk to human health compared to regulated HALs. This paper reviews the current research progress on the formation and transformation, cytotoxicity and genotoxicity of HALs in drinking water, and looks forward to the problems that should be paid attention in the future toxicological research of HALs in order to support the development of scientific drinking water standards.

Effect of Ageratina adenophora on hepatic and pulmonic pathological lesions in horses.

The effect of Ageratina adenophora on pathological characteristics of the liver and lungs as well as serum biochemical parameters in horses were investigated. Ten horses without ingestion history of Ageratina adenophora were classified into the control group, and 10 poisoned but survived horses with 3 months ingestion history were set as the case group. Results showed that serum AST, ALT, ALP, magnesium and phosphorus were elevated significantly, while creatinine was decreased remarkably. Hematoxylin and eosin staining of liver tissues showed diffuse swelling or destruction of hepatocytes, narrowing or atrophy of the hepatic sinusoids, and little lymphocytic infiltration; lung tissues presented destroyed alveoli and inflammatory cell infiltration.

[Study on the precision of the robot-assisted sacroiliac screw placement].

Objective: To evaluate the precision of the robot-assisted sacroiliac screw placement for posterior pelvis injury and the impacting factors. Methods: The clinical data of twenty-four cases of posterior pelvic fractures treated by percutaneous sacroiliac screw placement in Yantai shan Hospital from August 2016 to May 2018 were studied retrospectively. There were 17 males and 7 females with a mean age of 44.1 years (ranged from 17 to 71 years). According to AO classification, 17 cases were type B fractures (9 cases of type B1 and 8 type B2), and 7 cases were type C factures (3 cases of type C1, 2 type C2 and 2 type C3). All cases were treated by robot-assisted percutaneous sacroiliac screw placement (AO cannulated screws with a diameter of 7.3 mm). In the posterior pelvic surgeries for the 24 cases, 26 S(1) sacroiliac screws fixations and 18 S(2) sacroiliac screw fixations were placed in total, with single S(1) segmental fixation in 8 cases, single S(2) segmental fixation in 3 cases, S(1) and S(2) combined fixation in 13 cases, S(1) unidirectional one-sided fixation in 18 cases, S(1) bidirectional two-sided fixation in 3 cases, S(2) unidirectional one-sided fixation in 14 cases, S(2) bidirectional two-sided fixation in 2 cases and S(1) unidirectional double screws fixation in 2 cases. X-ray and CT examinations were taken for all 24 cases after operation. The visual analogue scale (VAS) of pain were performed before and after the operation. Results: All the sacroiliac screws were successfully implanted at once as planned with the assistance of the robot. The postoperative X-ray films and CT showed that none of the sacroiliac screws broke through the sacral body and the contralateral sacral wing's frontal cortex nor did they stray into the sacral canal and the intervertebral space. In 3 cases, the sacroiliac screws went closely against and wore out the front edge of iliac cortical density line and sacral alar slope and finally re-entered the sacral body. In 3 cases, sacroiliac screws touched upon the sacral nerve canals but did not break through the nerve canals. The mean VAS of pain was improved from 7.1 points (4-10 points) before the operation to 1.9 points (0-3 points) after. Conclusions: The robot-assisted sacroiliac screw placement shows high precision, and hence is worthy of clinical promotion; however the primary role of the surgeon could not be replaced.

KLF11 protects chondrocytes via inhibiting p38 MAPK signaling pathway.

OBJECTIVE: The purpose of this study was to explore the effects of Kruppel like factors 11 (KLF11) on oxidative stress, apoptosis, and endoplasmic reticulum stress (ERS) in osteoarthritis (OA) and its mechanism. PATIENTS AND METHODS: Human articular cartilage tissue was used to study the correlation between KLF11 and OA. Furthermore, human chondrocytes were used to explore the effects of KLF11 on oxidative stress, apoptosis, and ERS in chondrocytes by overexpressing KLF11 and using the OA inducer IL-1ß. The p38MAPK signaling pathway agonist P79350 was used to study the effect of KLF11 on the p38 MAPK signaling pathway. RESULTS: Articular cartilage tissue in OA patients and IL-1ß-induced chondrocytes expressed higher KLF11. Overexpression of KLF11 significantly reduced oxidative stress levels, apoptosis levels, and activity of ERS-related pathways in chondrocytes. Moreover, P79350 attenuated the protective effect of KLF11 on chondrocytes by activating the p38MAPK signaling pathway. CONCLUSIONS: KLF11 protects against OA by inhibiting oxidative stress, apoptosis, and ERS in chondrocytes by inhibiting p38MAPK signaling pathway.

[Concern about lung damage caused by guanidine cationic disinfectants].

This article summarized the use of guanidine disinfectants in China and the use of guanidine cationic disinfectants, polyhexamethylene guanidine (PHMG), in South Korea, which had caused severe lung damage events such as pulmonary fibrosis. The authors reviewed the studies that Chinese scientists employed ultrasonic atomization technology to simulate the actual scenario of human exposure to PHMG and proved the findings that PHMG could cause pulmonary fibrosis. These results could highlight the necessity of full attention to lung damage caused by guanidine disinfectants and its mechanism, so as to provide the important scientific basis for the protection of public health safety and the formulation of corresponding policies.

Biosynthetic potential of culturable bacteria associated with Apostichopus japonicus.

AIMS: Isolating culturable bacteria associated with sea cucumber (Apostichopus japonicus) and investigating their potential bioactivities are important approaches to discover natural marine products. METHODS AND RESULTS: A total of 161 isolates were obtained from sea cucumber collected along the Weihai coast of the North Yellow Sea, China. Identification and phylogenetic analysis of 61 isolates were conducted by 16S rRNA gene sequencing. The isolates belonged to 13 genera from 10 families in three phyla, including Firmicutes, Actinobacteria and Proteobacteria. The antimicrobial activities of all strains were determined using six indicator strains. Of the 161 isolates, 93 showed antibacterial activities against at least one of the indicator strains. The 26 strains with the strongest inhibitory effects were selected for screening the biosynthetic gene clusters of polyketide synthase (PKS-I, -II) and nonribosomal peptide synthetase (NRPS). Phylogenetic trees based on the amino acid sequences of the PKS or NRPS genes were constructed. Eleven strains with PKS genes and four strains with NRPS genes were detected. CONCLUSIONS: The data reveal the diversity of culturable bacteria associated with A. japonicus. Most strains showed broad-spectrum antimicrobial activities, and some strains with antimicrobial activities possessed PKS and NRPS genes. SIGNIFICANCE AND IMPACT OF THE STUDY: The results suggest that culturable bacteria associated with A. japonicus may act as a promising source of bioactive substances.

[Association between fruit consumption and stroke in adults in Qingdao: a prospective study].

Objective: To understand the fruit consumption of adults of Qingdao and examine the association between fruit consumption and stroke. Methods: We analyzed baseline data and stroke incidence of the participants who were aged 30-79 years and had been enrolled into the China Kadoorie Biobank (CKB) study from Qingdao city. Cox proportional hazards regression model was conducted to estimate the association of fruit consumption with risk of stroke. Results: A total of 35 509 participants were investgated in the baseline survey. Ratio of male to female was 1∶1.27, and the average age was (50.3±10.2) years. Respondents with higher frequency of fruit consumption were younger, more women, with higher education level and higher income (P<0.05). A total of 1 011 new cases of stroke were observed, with a stroke incidence of 387.63/100 000 person-years. Multivariate Cox regression analysis showed that fruit consumption had a protective effect on stroke incidence. Compared to the respondents who never consumed fruit, respondents who consumed fruit more than 4 days per week had a 44% lower risk of stroke incidence (HR=0.56, 95%CI: 0.50-0.62, P<0.05), and the risk reduced by 46% (HR=0.54, 95%CI: 0.46-0.64, P<0.05) and 42% (HR=0.58, 95%CI: 0.52-0.69, P<0.05) in male and female, respectively. Further adjustment for WC, BMI, SBP and random blood glucose did not change the association. Conclusion: Increasing fruit consumption can effectively decrease the risk of stroke. People should increase fruit consumption advisably to set up reasonable and healthy dietary habits.

[Research progress on exposure levels and toxic pathways of typical persistent organic pollutants in foods].

Dioxins, polybrominated diphenyl ethers, and benzo(a)pyrene are common organic pollutants in food. They have been of concern to academics and government administrations due to high residue and persistence, easy accumulation and strong harmful effects. The National Research Council of the United States of America published Toxicity Testing in the 21st Century: A Vision and Strategy in 2007, which proposed a new concept of toxicity testing that toxicity testing should take full consideration of population exposure data and base on in vitro tests, human cell lines, toxicity pathways and high-throughput screening. Meanwhile, systems biology, bioinformatics and rapid assay technologies will be used to better understand toxicity pathways-the cellular response pathways that can lead to adverse health effects when sufficient perturbing induced by chemicals exposure. The new toxicity testing strategy has changed the traditional testing pattern and has brought a wide impact on the international relevant fields. The European Union, the World Health Organization, and the United States Environmental Protection Agency, the Food and Drug Administration, and the National Center for Toxicological Research have organized relevant discussions and exploratory studies to address the new toxicity testing concept and how to evaluate and utilize the results of traditional toxicity test researches. Compared to the discussion, 'whether to do it', ten years ago, the question, 'how to do it', has become the concern of the current discussion. Therefore, how to respond to the concept of toxicity testing and how to effectively utilize and excavate traditional toxicity test data have been the focus of multi-disciplines and interdisciplinary academia such as toxicology, food hygiene and environmental science. Therefore, this article provides an overview of the exposure levels of dioxin, polybrominated diphenyl ethers and benzo[a]pyrene, which are typical persistent organic pollutants in food in China and the current research status of toxic pathways based on whole animal experiments. The exposure level, toxic effect and toxicity mechanism of three contaminants are analyzed and summarized in order to provide basis for future results based on the 21st century toxicity test compared with traditional tests and data mining analysis of these two kinds of data. Meanwhile, it also lays the foundation for the establishment of a toxicity testing framework based on exposure characteristics, toxic pathways, and biomarkers.

Metformin inhibits LPS-induced inflammatory response in VSMCs by regulating TLR4 and PPAR-γ.

OBJECTIVE: This study aims to explore whether the inhibitory role of metformin could inhibit LPS-induced inflammatory response in vascular smooth muscle cells (VSMCs) and its underlying mechanism. MATERIALS AND METHODS: VSMCs were extracted from aorta of Sprague Dawley rats. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay was performed to detect VSMCs viability after treatment with different concentrations of metformin. Levels of monocyte chemoattractant protein-1 (MCP-1), interleukin 6 (IL-6) and tumor necrosis factor-α (TNF-α) in VSMCs were detected by ELISA (enzyme-linked immunosorbent assay) and qRT-PCR (quantitative Real time-polymerase chain reaction). Protein and mRNA levels of toll like receptor 4 (TLR4) and peroxisome proliferators activated receptor γ (PPAR-γ) in VSMCs were detected by Western blot and qRT-PCR, respectively. Finally, VSMCs were treated with the PPAR-γ antagonist GW9662 and inflammatory indicators in cells were detected. RESULTS: No significant difference in VSMCs viability was found after 0-2 mM metformin treatment or 500 µg/L LPS induction for 24 h. After 500 µg/L LPS induction in VSMCs for 24 h, levels of MCP-1, TNF-α and IL-6 were remarkably elevated. Both mRNA and protein levels of TLR4 in VSMCs were upregulated after 500 µg/L LPS induction for 24 h, which were remarkably reversed by the treatment of different concentrations of metformin. Knockdown of TLR4 remarkably inhibited LPS-induced inflammatory response in VSMCs, manifesting as decreased levels of MCP1, TNF-α and IL-6, which were further downregulated after combination treatment of TLR4 knockdown and 20 mM metformin. Furthermore, both mRNA and protein levels of PPAR-γ in VSMCs were downregulated after 500 µg/L LPS induction for 24 h, which were remarkably reversed by the treatment of different concentrations of metformin. GW9662 treatment resulted in elevated expressions of MCP-1, TNF-α and IL-6, which were reversed by metformin treatment. CONCLUSIONS: Metformin can effectively inhibit the mRNA and protein expressions of IL-6, MCP-1, and TNF-α in LPS-induced VSMCs. The anti-inflammatory effects of metformin inhibit the inflammatory response through downregulating rely on the downregulation of TLR4 expression and upregulation ofng PPAR-γ activity.

Automated E-FRET microscope for dynamical live-cell FRET imaging.

Acceptor-sensitised 3-cube fluorescence resonance energy transfer (FRET) imaging (also termed as E-FRET imaging) is a popular fluorescence intensity-based FRET quantification method. Here, an automated E-FRET microscope with user-friendly interfaces was set up for dynamical online quantitative live-cell FRET imaging. This microscope reduces the time of a quantitative E-FRET imaging from 12 to 3 s. After locating cells, calibration of the microscope and E-FRET imaging of the cells can be performed automatically by clicking 'Capture' button on interfaces. E-FRET imaging was performed on the microscope for living cells expressing different FRET tandem constructs. Dynamical E-FRET imaging on the microscope for live cells coexpressing CFP-Bax and YFP-Bax treated by staurosporine (STS) revealed three Bax redistribution stages: Bax translocation from cytosol to mitochondria within 10 min, membrane insertion with conformational change on mitochondrial membrane within about 30 min, and subsequent oligomerisation within about 10 min. Because of excellent user-friendly interface and stability, the automated E-FRET microscope is a convenient tool for quantitative FRET imaging of living cell. LAY DESCRIPTION: Acceptor-sensitised 3-cube fluorescence resonance energy transfer (FRET) imaging (also termed as E-FRET) is a popular fluorescence intensity-based FRET quantification methods. E-FRET measurements are currently performed manually, and a complete FRET measurement takes about 12 s. E-FRET measurement necessitates not only a skilled operator and specialised equipment but also expertise in the interpretation of FRET signals, a considerable challenge in the application of FRET technology in living cells. Furthermore, manual E-FRET microscope is hard to perform dynamical quantitative FRET measurement, the ever-increasing applications in mapping the biochemical signal transduction within cells. Here, an automated E-FRET microscope with user-friendly interfaces was set up for dynamical online quantitative live-cell FRET imaging. This microscope reduces the time of a quantitative E-FRET imaging from 12 to 3 s. After locating cells, calibration of the microscope and E-FRET imaging of the cells can be performed automatically by clicking 'Capture' button on interfaces. Because of excellent user-friendly interface and stability, the automated E-FRET microscope is a convenient tool for quantitative FRET imaging of living cell.