Single-cell analysis reveals a unique microenvironment in peri-implantitis.

AIM: This study aimed to reveal the unique microenvironment of peri-implantitis through single-cell analysis. MATERIALS AND METHODS: Herein, we performed single-cell RNA sequencing (scRNA-seq) of biopsies from patients with peri-implantitis (PI) and compared the results with healthy individuals (H) and patients with periodontitis (PD). RESULTS: Decreased numbers of stromal cells and increased immune cells were found in the PI group, which implies a severe inflammatory infiltration. The fibroblasts were found to be heterogeneous and the specific pro-inflammatory CXCL13+ sub-cluster was more represented in the PI group, in contrast to the PD and H groups. Furthermore, more neutrophil infiltration was detected in the PI group than in the PD group, and cell-cell communication and ligand-receptor pairs revealed most neutrophils were recruited by CXCL13+ fibroblasts through CXCL8/CXCL6-CXCR2/CXCR1. Notably, our study demonstrated that the unique microenvironment of the PI group promoted the differentiation of monocyte/macrophage lineage cells into osteoclasts, which might explain the faster and more severe bone resorption in the progression of PI than PD. CONCLUSIONS: Collectively, this study suggests a unique immune microenvironment of PI, which may explain the differences between PI and PD in the clinic. These outcomes will aid in finding new specific and effective treatments for PI.

Comparison of different dosages of propofol combined with its equivalent alfentanil in outpatient abortion: a prospective, double-blinded, randomized trial.

OBJECTIVE: This study aimed at determining the optimal dose combination of alfentanil and propofol for outpatient abortion anesthesia. PATIENTS AND METHODS: The study was separated into two parts. In the first part, patients were to determine the median effective dose (ED50) and the 95% effective dose (ED95) of alfentanil in combination with 2.5 mg·kg-1 propofol to inhibit body movements during the abortion using the Dixon up-and-down sequential allocation method. In the second part, 170 patients were randomly divided into group C (2.0 mg·kg-1 propofol with alfentanil 12.16 µg·kg-1) and group E (2.5 mg·kg-1 propofol with its ED95) to compare the anesthetic effect. The primary outcome was the sedation level during general anesthesia. The secondary outcomes were circulation, respiratory complications, and postoperative recovery quality. RESULTS: The ED50 and the ED95 values of alfentanil were 3.37 µg·kg-1 (95% CI: 2.58-3.97 µg·kg-1) and 4.68 µg·kg-1 (95% CI: 4.04-9.32 µg·kg-1). The frequency of deep sedation in group E was significantly higher than in group C (76.5% vs. 60%). Patients in group C showed more wakefulness even during the surgery (14.3% vs. 4.4%). The results of our exploratory analyses did not reveal differences in respiratory depression, circulatory depression, postoperative side effects, or recovery outcomes. CONCLUSIONS: The combination of 2.5 mg·kg-1 propofol and 4.68 µg·kg-1 alfentanil produces a better sedative effect than the combination of 2.0 mg·kg-1 propofol and 12.16 µg·kg-1 alfentanil without increasing additional risks associated with anesthesia.

[Investigate the role of neutrophil extracellular traps in immune checkpoint inhibitor-associated myocarditis with programmed death protein-1 inhibitors involvement].

Objective: To investigate the role of neutrophil extracellular traps (NETs) in immune checkpoint inhibitor-associated myocarditis (ICIAM) with programmed death protein-1 (PD-1) inhibitors involvement, and to explore the therapeutic potential of targeting NETs in the treatment of ICIAM. Methods: Thirty 6-week-old male BALB/c mice were randomly divided into control group (n=10), myocarditis group (n=10), and treatment group (n=10). Apart from the control group, each mouse was subcutaneously injected with 100 µl of complete Freund's adjuvant containing 250 µg of mouse cardiac troponin I peptide on the 1st and 7th day. Starting on the 8th day, PD-1 inhibitor (15 µg/per mouse) was intraperitoneally injected every other day for a total of 5 times. Since 1 day before the beginning of PD-1+TnI injection, the treatment group was injected with PF-1355 (50 mg·kg-1·d-1) for 16 consecutive days. The mice's general state was observed during the whole process. Real-time fluorescence quantitative PCR (RTFQ-PCR) was carried out to evaluate the transcriptional regulation of neutrophil related chemokines, NETs, pyronecrosis related factors and proinflammatory cytokines. Immunohistochemistry, immunofluorescence and western blot were applied to determine the changes of pyrosis related molecules. Echocardiography showed the differences of main cardiac indexes while cardiac pathology compared the degree of inflammatory infiltration in 3 gruops. Results: The immunofluorescence intensity of myocardial NETs in the myocarditis group was significantly increased compared to the control group mice (2.49±0.08 and 0.99±0.26, P<0.001). The protein expression levels of pyroptosis-related NLRP3, cleaved-Caspase 1, Caspase 1, cleaved-GSDMD, GSDMD, IL-1ß and IL-18 in myocardial tissue of the model group were higher than those of the control group (all P<0.05). After treatment with PF-1355, compared to the myocarditis group, the left ventricular ejection fraction (LVEF) (73.58%±5.31% and 58.12%±3.19%, P<0.001) and left ventricular fraction shortening (LVFS) (39.78%±4.31% and 33.89%±2.19%, P<0.001) increased. H-E staining showed a reduction in inflammatory infiltration area in the treatment group compared to the myocarditis group (30.12%±3.57% and 14.92%±2.46%, P<0.001). The immunofluorescence intensity of NETs decreased in the treatment group compared to the myocarditis group (2.52±0.04 and 1.03±0.05, P<0.001). The levels of NLRP3 and other pyroptosis-related molecules were downregulated in the treatment group compared to the myocarditis group (all P<0.05). Conclusions: NETs lead to myocardial cell pyroptosis by activating the NLRP3 inflammasome in PD-1 inhibitor-associated myocarditis. The specific MPO inhibitor PF-1355 shows a therapeutic potential by regulating the formation of NETs, decreasing NLRP3 level and relieving myocardial pyroptosis, thus reducing myocardial damage.

[Analysis of the efficacy and influencing factors of sodium channel blockers in the treatment of focal epilepsy in infants under 6 months of age].

Objective: To analyze the efficacy and safety of the sodium channel blockers (SCB) antiseizure medication in the treatment of focal epilepsy in infants under 6 months of age. Methods: This was a case series study. Infants with focal epilepsy with onset within 6 months of age and treated with SCB attending the Department of Neurology of Beijing Children's Hospital from June 2016 to April 2022 were collected. The clinical data, auxiliary examinations, SCB application, efficacy, adverse reactions, and prognosis were analyzed retrospectively. Patients were grouped according to type of seizure and epileptic syndrome, age of onset and etiology. Chi square test and Fisher exact test were used to analyze the differences between groups statistically. Results: A total of 118 infants were enrolled, 65 males and 53 females, with an age of epilepsy onset of 56 (4, 114) days. Developmental and epileptic encephalopathy was diagnosed in 60 infants, 39 had self-limited neonatal and (or) infantile epilepsy, and 19 had non-syndromic focal epilepsy. Application of SCB: 106 used oxcarbazepine, 2 used lacosamide, 9 switched from oxcarbazepine to lacosamide or a combination of 2 SCB, and 1 used oxcarbazepine, lacosamide, and lamotrigine successively; oxcarbazepine was the first choice in 46 cases. The age at which SCB was applied was 103 (53, 144) days. The children were followed up for 6 months to 6 years. SCB was effective in 89 cases (75.4%), including 70 cases (59.3%) who achieved seizure freedom. The seizure-free rate was higher in the focal epilepsy only group than in the group with other seizure types (64.4% (65/101) vs. 4/17, χ²=9.99, P<0.05). The responder and seizure-free rates were all higher in the group with the onset age of >3-6 months than the group >1-3 months (84.4% (38/45) vs. 62.5% (20/32), 73.3% (33/45) vs. 46.9% (15/32), χ²=4.85 and 5.58, both P<0.05). With the exception of variants in the PRRT2 gene, those with variants in sodium or potassium channels had higher responder and seizure-free rates than those with variants in other genes(86.2% (25/29) vs. 45.5% (10/22), 62.1% (18/29) vs. 22.7% (5/22), χ²=9.65 and 7.82,both P<0.05). The most common adverse event was transient hyponatremia, which happened in 66 cases (55.9%). There were 9 cases of rash, which subsided in 6 cases after discontinuing oxcarbazepine and switching to lacosamide, and 7 cases of electrocardiogram abnormalities, which improved after withdrawing oxcarbazepine and changing to lacosamide in 1 case. Conclusion: SCB are effective and tolerable in the treatment of focal epilepsy in infants under 6 months of age, with better efficacy in patients with genetic variants of the sodium or potassium channel, focal seizures only, and seizure onset >3-6 months of age.

[Application of liver venous deprivation in secondary hepatic resection of primary liver cancer].

Objective: To investigate the efficacy and safety of liver venous deprivation (LVD) before secondary resection of primary liver cancer. Methods: 56 patients with advanced primary liver cancer who were not suitable for primary resection in Liver Surgery Department of Xinxiang Central Hospital from January 2018 to January 2019 were analyzed retrospectively. They were divided into liver vein deprivation group (LVD group: LVD+ PVE, n=26) and portal vein embolization group (PVE group, n=30). The dynamic changes of liver reserve function and future liver remnant volume (FLR-V), R0 resection rate, surgical complications, postoperative recurrence rate and overall survival rate of two groups before and after LVD/PVE were compared. Results: The success rate of puncture and embolization in LVD group and PVE group was 100%. There were no grade Ⅳ complications, and there was no significant difference of grades Ⅰ, Ⅱ and Ⅲ complications between the groups (P=0.808). The FLR-V of LVD group before embolization, 7, 14 and 21 days after embolization was (493.1±25.8), (673.2±56.1), (779.5±81.6) and (853.3±85.2) cm(3), respectively. The FLR-V of PVE group before embolization, 7, 14 and 21 days after embolization were (502.4±20.1), (688.6±43.9), (656.8±73.7) and (563.5±69.1) cm(3), respectively. There was no significant difference in FLR-V between the two groups before and 7 days after embolization (P>0.05). The FLR-V of LVD group was higher than that of PVE group at 14 and 21 days after embolization (P<0.01). The preparation time of LVD group was (20.4±6.3) days, which was shorter than that of PVE group [(31.5±8.8) days, P=0.045]. The rate of secondary hepatectomy was 92.3% (24/26), which was higher than that of PVE group [70.0% (21/30), P=0.036]. The R0 resection rate was 87.5% (21/24), which was higher than that of the PVE group [57.1% (12/21), P=0.022]. However, there were no significant differences in surgical methods, operation time, intraoperative blood loss, Clavien-Dindo complication grade and length of hospital stay between the two groups (P>0.05). After hepatectomy, the median recurrence time and median survival time of LVD group were 12.6 months and 21.3 months, respectively, which were longer than those of PVE group (9.4 months and 13.5 months, respectively, P<0.01). Conclusions: For patients with advanced liver cancer who are not suitable for primary hepatectomy, preoperative LVD can significantly increase FLR-V, improve the resection rate of secondary surgery, shorten the preparation time of two operations, and do not increase surgical complications. Moreover, patients with LVD can improve the R0 resection rate of secondary surgery. The postoperative recurrence time and overall survival rate of patients with LVD are better than those of patients with PVE, and LVD has a good long-term effect.

[Evaluation of left ventricular function with left atrio-ventricular longitudinal strain in patients with lymphoma underwent anthracycline therapy].

Objective: To analyze the value of 3-dimensional speckle tracking echocardiograghy (3D-STE) derived strain parameters on the detection of subclinical myocardial deformation alterations in patients with lymphoma treated with anthracycline agents. Methods: This study was a retrospective study. A total of 37 patients with newly diagnosed diffuse large B cell non-Hodgkin lymphoma between December 2012 and December 2014 in Cancer Center, Fudan university were included. 3D-STE strain measurements were performed at baseline (T0),after the completion of two therapy circles (T1) and at the end of anthracycline regimen chemotherapy (Te). Echocardiography images were analyzed on the TTA workstation, and the indexes included left atrial minimum volume (LAVmin), left atrial emptying index (LAEF), left atrial active emptying index (LAAEF), as well as the left ventricular global longitudinal strain (LVGLS), left ventricular global circumferential strain (LVGCS), left atrial global longitudinal strain (LAGLS). The overall left atrioventricular longitudinal strain (LAVGLS) was calculated, which was the sum of the absolute values of LVGLS and LAGLS. The changes of left ventricular strain indexes measured by 3D-STE at different time points of patients were evaluated. Results: Thirty-seven patients with DLBCL, aged (48.3±12.1)years, including 23 males (63.9%), were enrolled. Compared with baseline, LVGLS (T1: (-18.63±4.73)% vs. (-22.13±4.40)%, P=0.001; Te:(-18.26±4.64)% vs. (-22.13±4.40)%, P<0.001), LAGLS (T1: (20.41±5.56)% vs. (23.98±5.59)%, P=0.003; Te: (17.60±3.96)% vs. (23.98±5.59)%, P<0.001) and LAVGLS (T1: (39.05±7.60)% vs. (46.11±7.77)%, P<0.001; Te: (40.34±8.55)% vs. (46.11±7.77)%, P<0.001) were all deteriorated at the T1 and Te. While LVGCS ((-21.98±5.82)% vs. (-26.15±7.51)%, P=0.010), LAVmin ((23.93±7.29)ml vs. (20.33±7.03)ml, P=0.029), LAEF ((28.94±11.16)% vs. (35.79±11.12)%, P=0.002) and LAAEF ((11.93±10.00)% vs. (18.10±9.96)%, P=0.013) were decreased only until Te. Conclusions: 3D-STE strain measurements could detect early myocaridial function alteration in patients receiving anthracycline regimen chemotherapy, thus may provide a novel approach to monitor anthracycline caused myocardial toxicity.

Comparison of different colistin regimens for the treatment of pneumonia caused by multidrug-resistant microorganisms: a systematic review and meta-analysis.

OBJECTIVE: Multidrug-resistant pneumonia is a common cause of hospital-related morbidity and mortality across the world. The high prevalence of multidrug-resistant pneumonia due to resistant gram-negative pathogens has led to a re-introduction of colistin. The adverse events associated with intravenous colistin can be alleviated by administering the drug nasally (i.e., inhalation) or in a combination including both inhalation and intravenous presentations of the drug. A review study compared the impact of these administration methods on clinical, morbidity, and mortality-related outcomes in patients with multiple-drug resistant pneumonia. However, the publication of newer cohort trials, warrants an update of the state of the evidence. To compare the clinical, morbidity, and mortality outcomes in patients with multidrug-resistant pneumonia receiving either intravenous colistin or combined drug presentations (ie, inhaled and intravenous). MATERIALS AND METHODS:  A systematic search of the academic literature was performed according to the PRISMA guidelines across five databases (Web of Science, EMBASE, CENTRAL, Scopus, and MEDLINE). We conducted a random-effect meta-analysis to compare outcomes such as rate of clinical cure, microbiological eradication, nephrotoxicity, and overall mortality in patients with multidrug-resistant pneumonia receiving either intravenous colistin, inhaled colistin, or a combination of those administration routes. RESULTS:  From 963 studies, we found 16 eligible studies with 1651 patients (61.6 ± 7.7 years) with multidrug-resistant pneumonia who had received either intravenous, inhaled colistin or a combined inhaled/intravenous administration. Our meta-analysis revealed higher rates of clinical cure (OR, 1.61) and microbiological eradication (1.37) in patients receiving combined intravenous/inhaled colistin than in those receiving intravenous colistin alone. Additional analyses revealed higher rates of nephrotoxicity (1.30) and mortality (1.44) in patients receiving intravenous colistin than in those receiving combined intravenous/inhaled colistin. CONCLUSIONS:  We provide evidence showing improved clinical, morbidity, and mortality outcomes in patients with multidrug-resistant pneumonia receiving inhaled colistin or combined inhaled/intravenous colistin than those receiving intravenous colistin alone. These findings should help clinicians stratify the risks associated with different colistin administration routes to manage multidrug-resistant pneumonia.

[Accuracy and clinical outcome of a real-time surgical navigation system for the placement of quad zygomatic implants].

Objective: To evalute the accuracy and clinical outcome of a real-time navigation system for the placement of quad zygomatic implants. Methods: Twenty-four patients [9 males and 15 females, mean age was (50.8±14.7) years old], from January 2015 to December 2019, with 96 zygomatic implants placed under a real-time navigation system in Department of Second Dental Center and Department of Oral Implantology of Ninth People's Hospital, Shanghai Jiaotong University School of Medicine were included in the study. The preoperative and the postoperative multislice CT or cone-beam CT were fused to measure and record the entry, exit and angle deviation between the planned and placed implants. The implants were divided into groups according to implant insertion approach (real-time navigation and free-hand), implant length (<47.5 mm and ≥47.5 mm) and implant position (proximal and distal implant). And the differences of implant accuracy were analyzed. The intraoperative and postoperative complications were also recorded. The implant survival rate was evaluated after 6 months follow-up. A P value<0.05 indicates statistical significance. Results: The mean entry, exit and angle deviation of zygomatic implants were (1.49±0.64) mm, [2.03(1.58, 2.40)] mm and (2.49°±1.12°), respectively. The average entry, exit and angle deviation of the navigation guided implant insertion group were (1.45±0.60) mm, (1.96±0.44) mm and (2.66±1.13°) respectively, while those of the free-hand group were (1.50±0.64) mm, (2.04±0.79) mm and (2.50°±1.13°) respectively. There was no significant difference between the two groups (P>0.05). The average entry, exit and angle deviation of the group with length<47.5 mm were (1.42±0.60) mm, (2.13±0.60) mm and (2.61°±1.08°) respectively and those of the group with length ≥ 47.5 mm were (1.52±0.65) mm, (1.98±0.82) mm and (2.43°±1.14°) respectively. No significant difference was found between the two groups (P>0.05). In proximal implant group, the average entry, exit and angle deviation were (1.55±0.69) mm, (2.05±0.92) mm and (2.48°±1.16 °) respectively while those of distal implant group were (1.43±0.57) mm, (2.01±0.57) mm and (2.49°±1.10°), respectively. No significant difference was detected between the two groups (P>0.05). All zygomatic implants were placed uneventfully. There were no intra-operative complications, and post-operative reversible complications developed in 3 patients. Two zygomatic implants were lost and the overall zygomatic implant survival rate was 97.9% (94/96) within a follow-up of 6 months. Conclusions: Quad zygomatic implant placement can be achieved with high accuracy and predictable clinical outcome under guidance of a real-time navigation system.

[Predictive value of two-dimentional shear wave elastography in posthepatoectomy liver failure].

Objective: To investigate the value of two-dimensional shear wave elastography (2D SWE) combined with clinical biochemical data in predicting posthepatoectomy liver failure (PHLF) in patients with hepatocellular carcinoma (HCC). Methods: A total of 274 HCC patients who underwent hepatectomy in Zhongshan Hospital Fudan University from January 2015 to January 2016 were retrospectively collected, including 235 males and 39 females, age 19-80 (56±11) years. All patients were confirmed to be HCC by postoperative pathology. The preoperative 2D SWE examination, laboratory examination results and intraoperative indicators were analyzed. According to the occurrence of PHLF after surgery, single factor analysis and multiple logistic regression analysis were performed on the above indicators to obtain a binary logistic regression model, and evaluate the diagnostic effect of the model on PHLF. In addition, 103 HCC patients from October 2019 to January 2020 were retrospectively collected as an external validation set, including 89 males and 14 females, age 23-80 (55±11) years old. Results: The liver stiffness measurement (LSM) obtained from 2D SWE, INR and Laminin (LN) were independent predictors of PHLF. The formula of prediction model PM=-15.451+0.095×LSM+11.7×INR+0.012×LN was obtained by combining above three factors. The area under the curve (AUC) of PHLF was 0.82, which was higher than that of end-stage liver disease model (MELD) score and Child-Pugh grading diagnosis of PHLF. The AUC of PHLF predicted by PM in the external validation group was 0.81. Conclusion: 2D SWE is helpful for clinicians to evaluate liver reserve function preoperatively and to predict the occurrence of PHLF in patients with HCC.

[Application and influencing factors of endorectal ultrasound in T-stage restaging of rectal cancer following neoadjuvant therapy].

Objective: To evaluate the accuracy and influencing factors of T-stage restaging of rectal cancer following neoadjuvant therapy with endorectal ultrasonography (ERUS). Methods: In a retrospective study, endorectal ultrasound was performed in 86 patients with rectal cancer following neoadjuvant therapy. The imaging results were compared with postoperative pathological T-stage. Results: The accuracy of overall T-stage restaging with ERUS was 67.4% (58/86). Additionally, the accuracy of restaging in middle and high rectal cancer was higher, with an accuracy of 76.1%(35/46)and 100%(4/4) respectively. Univariate analysis showed that the location of tumors was an independent factor affecting the accuracy of ERUS(P=0.033). Conclusion: ERUS is an effective method to restage T-stage of rectal cancer following neoadjuvant therapy.

Rapid detection of equine infectious anaemia virus nucleic acid by insulated isothermal RT-PCR assay to aid diagnosis under field conditions.

BACKGROUND: Control of equine infectious anaemia (EIA) currently depends on serological diagnosis of infected equids. However, recently infected equids may not produce detectable anti-EIAV antibodies up to 157 days post infection and so present a high transmission risk. Therefore, direct nucleic acid detection methods are urgently needed to improve EIAV surveillance and management programs in counties where the disease is endemic. OBJECTIVES: To evaluate a field-deployable, reverse transcription-insulated isothermal PCR (RT-iiPCR) assay targeting the conserved 5' untranslated region (5' UTR)/exon 1 of the tat gene of EIAV. STUDY DESIGN: The analytical and clinical performance of the newly developed EIAV RT-iiPCR was evaluated by comparison with a EIAV real-time RT-PCR (RT-qPCR) along with the AGID test. METHODS: Analytical sensitivity was determined using in vitro transcribed RNA containing the target area of the 5' UTR/tat gene and samples from two EIAV-positive horses. Specificity was verified using nine common equine viruses. Clinical performance was evaluated by comparison with EIAV RT-qPCR and AGID using samples derived from 196 inapparent EIAV carrier horses. RESULTS: EIAV RT-iiPCR did not react with other commonly encountered equine viruses and had equivalent sensitivity (95% detection limit of eight genome equivalents), with a concordance of 95.41% to conventional EIAV RT-qPCR. However, the RT-qPCR and RT-iiPCR had sensitivities of 43.75 and 50.00%, respectively, when compared to the AGID test. MAIN LIMITATIONS: Low viral loads commonly encountered in inapparent EIAV carriers may limit the diagnostic sensitivity of RT-PCR-based tests. CONCLUSIONS: Although EIAV RT-iiPCR is not sufficiently sensitive to replace the current AGID test, it can augment control efforts by identifying recently exposed or "serologically silent" equids, particularly as the latter often represent a significant transmission risk because of high viral loads. Furthermore, the relatively low cost and field-deployable design enable utilisation of EIAV RT-iiPCR even in remote regions.

[Development and progress of sixty years and stepping firmly to the future].

Since its establishment for 60 years, Department of Burns of the Second Affiliated Hospital of Zhejiang University School of Medicine has grown into a famous regional burn center in China under the leading of the pioneers and through the efforts of several generations. The department has distinctive disciplinary features in burn care, nutritional support, scar prevention and treatments, standard management of chronic wound, and skin tissue engineering research, making positive contribution to the development of burn medicine in China.

On-site detection of equid alphaherpesvirus 3 in perineal and genital swabs of mares and stallions.

Equine coital exanthema (ECE) is an infectious, venereally transmitted muco-cutaneous disease affecting mares and stallions, caused by equid alphaherpesvirus 3 (EHV3). Diagnostic tools for rapid identification of EHV3 are of primary importance to diminish the risk of EHV3 dissemination at the time of breeding. In the last years, it has been shown that the performance of the insulated-isothermal polymerase chain reaction (iiPCR) is comparable to virus isolation, nested PCR and real-time PCR (qPCR) in detecting pathogens of various animal species. Analytical sensitivity and specificity of the iiPCR were compared with a qPCR, using a plasmid containing the target region of the EHV3 glycoprotein G gene and an Argentinian EHV3 isolate (E/9283/07 C3A). In order to evaluate the diagnostic performance of the iiPCR, nucleic acids of 85 perineal and genital swabs (PGS) of mares and stallions were extracted by tacoTM mini and tested by both techniques. EHV3 was detected in 46 and 45 of the 85 PGS by the iiPCR and qPCR, respectively. There was almost perfect agreement between the two diagnostic methods (98.82%; 95% CI: 95.03-100%; κ = 0.98). The iiPCR had a limit of detection of 95.00% at 6 genome equivalents per reaction and a detection endpoint for viral DNA comparable to that of the qPCR, and did not react with six non-targeted equine pathogens. The iiPCR represents a sensitive and specific method for the rapid on-site diagnosis of EHV3 infection. Its routinely implementation in breeding facilities, and artificial insemination and embryo transfer centers, will contribute to prevent the dissemination of this venereal, highly contagious disease in horses.

Effect of pH on uranium(VI) biosorption and biomineralization by Saccharomyces cerevisiae.

Biosorption of radionuclides by microorganisms is a promising and effective method for the remediation of contaminated areas. pH is the most important factor during uranium biosorption by Saccharomyces cerevisiae because the pH value not only affects the biosorption rate but also affects the precipitation structure. This study investigated the effect of pH on uranium (VI) biosorption and biomineralization by S. cerevisiae. Cells have the ability to buffer the solution to neutral, allowing the biosorption system to reach an optimal level regardless of the initial pH value. This occurs because there is a release of phosphate and ammonium ions during the interaction between cells and uranium. The uranyl and phosphate ions formed nano-particles, which is chernikovite H2(UO2)2(PO4)2·8H2O (PDF #08-0296), on cell surface under the initial acidic conditions. However, under the initial alkaline conditions, the uranyl, phosphate and ammonium ions formed a large amount of scale-like precipitation, which is uramphite (NH4)(UO2)PO4·3H2O (PDF #42-0384), evenly over on cell surface.

Activation of the STING-IRF3 pathway promotes hepatocyte inflammation, apoptosis and induces metabolic disorders in nonalcoholic fatty liver disease.

BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) is a common result of obesity and metabolic syndrome. Hepatocyte injury and metabolic disorders are hallmarks of NAFLD. Stimulator of interferon genes (STING) and its downstream factor interferon regulatory factor 3 (IRF3) trigger inflammatory reaction in response to the presence of cytosolic DNA. STING has recently been shown to play an important role in early alcoholic liver disease. However, little is known about the role of STING-IRF3 pathway in hepatocyte injury. Here, we aimed to examine the effect of STING-IRF3 pathway on hepatocyte metabolism, inflammation and apoptosis. METHODS: We examined the activation of the STING-IRF3 pathway, a high-fat diet (HFD)-induced obese mouse model, and determined the role of this pathway in a free fatty acid (FFA)-induced hepatocyte inflammatory response, injury, and dysfunction in L-O2 human liver cells. RESULTS: STING and IRF3 were upregulated in livers of HFD-fed mice and in FFA-induced L-O2 cells. Knocking down either STING or IRF3 led to a significant reduction in FFA-induced hepatic inflammation and apoptosis, as evidenced by modulation of the nuclear factor κB (NF-κB) signaling pathway, inflammatory cytokines, and apoptotic signaling. Additionally, STING/IRF3 knockdown enhanced glycogen storage and alleviated lipid accumulation, which were found to be associated with increased expression of hepatic enzymes in glycolysis and lipid catabolism, and attenuated expression of hepatic enzymes in gluconeogenesis and lipid synthesis. CONCLUSIONS: Our results suggest that the STING-IRF3 pathway promotes hepatocyte injury and dysfunction by inducing inflammation and apoptosis and by disturbing glucose and lipid metabolism. This pathway may be a novel therapeutic target for preventing NAFLD development and progression.

[Efficacy and safety of linezolid among patients with methicillin-resistant Staphylococcus aureus bacteremia].

Objective: To study the efficacy and safety of linezolid for the treatment of patients with bacteremia caused by methicillin-resistant Staphylococcus aureus (MRSA). Methods: Totally 52 cases of MRSA bacteremia patients, from January 2010 to April 2014 in the First Affiliated Hospital, School of Medicine, Zhejiang University, were retrospectively analyzed. They were classified into two groups based on linezolid therapeutic regimen: primary treatment with linezolid (19 cases) and alternated to linezolid (33 cases). The following data were collected and compared: clinical characteristics, lasting time of fever, bacterial clearance rate, clinical efficacy, fatality rate, and adverse events. Results: Forty three of the 52 patients (82.7%) suffered complicated MRSA bacteremia. The most common clinical feature was fever[86.5%(45/52)]. Linezolid was initiatively used mostly because of renal insufficiency[68.4%(13/19)]. In the other 33 patient, glycopeptides were initiatively used, then alternated to linezolid because of persistent fever[69.7%(23/33)]; damage of kidney function during treatment period of glycopeptides[12.1%(4/33)]; occurrence of new infectious site related to MRSA[18.2%(6/33)]. The clinical efficacy were 78.9%(15/19) in the group of primary treatment with linezolid and 81.8% (27/33) in the group of alternated to linezolid, persistent time of fever were 4(3, 15) d and 12(5, 24) d, mortality during 28 d period were 15.8% (3/19) and 9.1% (3/33), adverse rate were 15.8% (3/19) and 12.1% (4/33) in these two groups, respectively (all P>0.05). Conclusion: Linezolid is an option with high clinical efficacy and good safety for MRSA bacteremia patients.

[Value of dual contrast-enhanced ultrasound in the preoperative T staging of rectal carcinoma].

Objective: To explore the value of dual contrast-enhanced ultrasound in preoperative T staging of rectal carcinoma. Methods: Dual contrast-enhanced ultrasound examinations were performed on 72 patients with rectal carcinoma via transrectal infusion and intravenous injection. The accordance of preoperative dual contrast-enhanced ultrasound results and postoperative pathologic results was evaluated retrospectively. Results: The overall accordance rate of preoperative T staging was 73.6% (53/72). And accordance rate was 100.0% (3/3), 100.0% (5/5), 68.4% (13/19), 71.4% (25/35)and 70.0% (7/10) for Tis , T1, T2, T3 and T4, respectively. The consistency was good (κ=0.607, χ(2) =8.363, P<0.01). The accordance rate of middle/lower vs high rectal carcinoma was 68.7% and 85.7%. Conclusion: Dual contrast-enhanced ultrasound can provide reference for preoperative T staging for patients with rectal carcinoma.

Induction of endoplasmic reticulum stress by deletion of Grp78 depletes Apc mutant intestinal epithelial stem cells.

Intestinal epithelial stem cells are highly sensitive to differentiation induced by endoplasmic reticulum (ER) stress. Colorectal cancer develops from mutated intestinal epithelial stem cells. The most frequent initiating mutation occurs in Apc, which results in hyperactivated Wnt signalling. This causes hyperproliferation and reduced sensitivity to chemotherapy, but whether these mutated stem cells are sensitive to ER stress induced differentiation remains unknown. Here we examined this by generating mice in which both Apc and ER stress repressor chaperone Grp78 can be conditionally deleted from the intestinal epithelium. For molecular studies, we used intestinal organoids derived from these mice. Homozygous loss of Apc alone resulted in crypt elongation, activation of the Wnt signature and accumulation of intestinal epithelial stem cells, as expected. This phenotype was however completely rescued on activation of ER stress by additional deletion of Grp78. In these Apc-Grp78 double mutant animals, stem cells were rapidly lost and repopulation occurred by non-mutant cells that had escaped recombination, suggesting that Apc-Grp78 double mutant stem cells had lost self-renewal capacity. Although in Apc-Grp78 double mutant mice the Wnt signature was lost, these intestines exhibited ubiquitous epithelial presence of nuclear ß-catenin. This suggests that ER stress interferes with Wnt signalling downstream of nuclear ß-catenin. In conclusion, our findings indicate that ER stress signalling results in loss of Apc mutated intestinal epithelial stem cells by interference with the Wnt signature. In contrast to many known inhibitors of Wnt signalling, ER stress acts downstream of ß-catenin. Therefore, ER stress poses a promising target in colorectal cancers, which develop as a result of Wnt activating mutations.

Galectin-1 induces hepatocellular carcinoma EMT and sorafenib resistance by activating FAK/PI3K/AKT signaling.

Galectin-1 (Gal-1) is involved in several pathological activities associated with tumor progression and chemoresistance, however, the role and molecular mechanism of Gal-1 activity in hepatocellular carcinoma (HCC) epithelial-mesenchymal transition (EMT) and sorafenib resistance remain enigmatic. In the present study, forced Gal-1 expression promoted HCC progression and sorafenib resistance. Gal-1 elevated αvß3-integrin expression, leading to AKT activation. Moreover, Gal-1 overexpression induced HCC cell EMT via PI3K/AKT cascade activation. Clinically, our data revealed that Gal-1 overexpression is correlated with poor HCC survival outcomes and sorafenib response. These data suggest that Gal-1 may be a potential therapeutic target for HCC and a biomarker for predicting response to sorafenib treatment.

Expression level of microRNA-195 in the serum of patients with gastric cancer and its relationship with the clinicopathological staging of the cancer.

OBJECTIVE: To study miroRNA-195 (miR-195) expression in the serum and cancer tissue of patients with gastric cancer and to investigate the relationship between its expression and clinicopathological features of gastric cancer. PATIENTS AND METHODS: Sixty-two patients with gastric cancer admitted to our institution were included in the study group, and 36 healthy persons undergoing health check-up at our institution served as control group. miR-195 expressions in the serum, gastric cancer tissue and corresponding paracancerous tissue in subjects of two groups were measured by using quantitative fluorescent real-time PCR (QF-RT-PCR), and the relationship between miR-195 and the clinicopathological features of the cancer was investigated. RESULTS: miR-195 expression level in the serum of gastric cancer patients was significantly lower than that in the control group (p <0.05). miR-195 expression in gastric cancer tissue was also significantly lower than that in corresponding paracancerous tissue (p <0.05). The results of correlation analysis showed that low expression of miR-195 was negatively correlated with the infiltration depth, the extent of differentiation, the clinical staging and lymph node metastasis, all with statistical significance (p <0.05), but not significantly correlated with tumor locations (p >0.05). CONCLUSIONS: Low expression of miR-195 in patients with gastric cancer may play a certain role in promoting the genesis and development of gastric cancer and it can function as a potential novel tumor marker for the early diagnosis and prognosis evaluation of gastric cancer.