The serine/threonine protein kinase of Streptococcus suis serotype 2 affects the ability of the pathogen to penetrate the blood-brain barrier.

Streptococcus suis serotype 2 (SS2) is a zoonotic agent that causes meningitis in humans and pigs. However, the mechanism whereby SS2 crosses the microvasculature endothelium of the brain is not understood. In this study, transposon (TnYLB-1) mutagenesis was used to identify virulence factors potentially associated with invasive ability in pathogenic SS2. A poorly invasive mutant was identified and was found to contain a TnYLB-1 insertion in the serine/threonine kinase (stk) gene. Transwell chambers containing hBMECs were used to model the blood-brain barrier (BBB). We observed that the SS2 wild-type ZY05719 strain crossed the BBB model more readily than the mutant strain. Hence, we speculated that STK is associated with the ability of crossing blood-brain barrier in SS2. In vitro, compared with ZY05719, the ability of the stk-deficient strain (Δstk) to adhere to and invade both hBMECs and bEnd.3 cells, as well as to cross the BBB, was significantly attenuated. Immunocytochemistry using antibodies against claudin-5 in bEnd.3 cells showed that infection by ZY05719 disrupted BBB tight junction proteins to a greater extent than in infection by Δstk. The studies revealed that SS2 initially binds at or near intercellular junctions and crosses the BBB via paracellular traversal. Claudin-5 mRNA levels were indistinguishable in ZY05719- and Δstk-infected cells. This result indicated that the decrease of claudin-5 was maybe induced by protein degradation. Cells infected by ZY05719 exhibited higher ubiquitination levels than cells infected by Δstk. This result indicated that ubiquitination was involved in the degradation of claudin-5. Differential proteomic analysis showed that E3 ubiquitin protein ligase HECTD1 decreased by 1.5-fold in Δstk-infected bEnd.3 cells relative to ZY05719-infected cells. Together, the results suggested that STK may affect the expression of E3 ubiquitin ligase HECTD1 and subsequently increase the degradation of claudin-5, thus enabling SS2 to traverse the BBB.

Virulent and Vaccine Strains of Streptococcus equi ssp. zooepidemicus Have Different Influences on Phagocytosis and Cytokine Secretion of Macrophages.

Swine streptococcosis is a significant threat to the Chinese pig industry, and Streptococcus equi ssp. zooepidemicus (SEZ) is one of the major pathogens. SEZ ATCC35246 is a classical virulent strain, while SEZ ST171 is a Chinese attenuated vaccine strain. In this study, we employed stable isotope labeling by amino acids in cell culture and liquid chromatography-mass spectrometry (LC-MS) to determine the differential response of macrophages to infection by these two strains. Eighty-seven upregulated proteins and 135 downregulated proteins were identified. The proteomic results were verified by real-time polymerase chain reaction for 10 chosen genes and Western blotting for three proteins. All differentially abundant proteins were analyzed for their Gene Ontology and Kyoto Encyclopedia of Genes and Genomes annotations. Certain downregulated proteins were associated with immunity functions, and the upregulated proteins were related to cytomembrane and cytoskeleton regulation. The phagocytosis rate and cytokine genes transcription in Raw264.7 cells during SEZ ATCC35246 and ST171 infection were detected to confirm the bioinformatics results. These results showed that different effects on macrophage phagocytosis and cytokine expression might explain the different phenotypes of SEZ ATCC35246 and ST171 infection. This research provided clues to the mechanisms of host immunity responses to SEZ ST171and SEZ ATCC35246, which could identify potential therapy and vaccine development targets.

SILAC and LC-MS/MS identification of Streptococcus equi ssp. zooepidemicus proteins that contribute to mouse brain microvascular endothelial cell infection.

Streptococcus equi ssp. zooepidemicus (SEZ) causes meningitis in both humans and animals. Some dissociative proteins of SEZ are cytotoxic to mouse brain microvascular endothelial cells (mBMECs) and may contribute to the penetration of SEZ across the blood-brain barrier (BBB). In this study, the ability of SEZ to penetrate across an in vitro BBB model was confirmed. We used stable isotope labeling with amino acids in cell culture (SILAC) to label SEZ proteins with heavy or light isotope-tagged amino acids, along with LC-MS/MS to determine which SEZ proteins were involved in interactions with mBMECs. The efficiency of SEZ protein isotope labeling was 94.7 %, which was sufficient for further analysis. Forty-nine labeled peptides were identified as binding to mBMECs, which matched to 25 SEZ proteins. Bioinformatic analysis indicated that most of these proteins were cytoplasmic. These proteins may have functions in breaching the host BBB, and some of them are known virulence factors in other bacteria. Indirect immunofluorescence results indicated that SEZ enolase had binding activity toward mBMECs. Protective test results showed that enolase was a protective antigen against SEZ infection. This research is the first application of SILAC combined with LC-MS/MS to identify SEZ proteins that may contribute to the infection of mBMECs and potentially show functions related to breaching the BBB. The outcomes provide many future avenues for research into the mechanism of SEZ-induced meningitis.

Analysis of therapeutic effect and influencing factors of radiofrequency ablation for colorectal cancer liver metastases / 中华放射学杂志

Objective:To explore the efficacy and influencing factors of radiofrequency ablation (RFA) in the treatment of colorectal cancer liver metastases (CRLM).Methods:The clinical and imaging data of 281 patients (477 intrahepatic metastatic tumors) who received percutaneous RFA treatment in Sir Run Run Shaw Hospital, Zhejiang University School of Medicine from December 2009 to December 2020 were retrospectively analyzed. Factors that may affect the efficacy of RFA were recorded, including carcinoembryonic antigen (CEA), differentiation, extrahepatic metastasis, tumor location and size, complications and other information. Patients were followed up through hospital admissions, telephone, etc. The primary endpoints were overall survival (OS) and local tumor progression-free survival (LTPFS). Univariate and multivariate logistic regression models were used to identify predictors of residual tumor. Univariate and multivariate Cox proportional hazards regression were used to identify the influencing factors of LTPFS and OS. The median LTPFS and OS were estimated by the Kaplan-Meier curve and compared by the log-rank test.Results:After RFA, 68 (14.3%) tumor residues were observed. Multivariate logistic regression showed that the risk factors for residual tumor were size ≥20 mm, high-risk and perivascular location, and minimal ablative margin<5 mm. During the follow-up period, the main complication rate was 4.3% (12/281) and the fatality rate was 31.3% (88/281). At the same time, local tumor progression was found in 167 (35.0%) lesions post-RFA. The median time of LTPFS and OS estimated by the Kaplan Meier method were 35.0 (95%CI 26.53-43.48) and 44.0 (95%CI 29.70-58.30) months, respectively. The cumulative proportion of LTPFS and OS were 37.2% and 40.4% respectively in the 5th year. Multivariate Cox proportional hazard regression showed that CEA≥30 ng/ml, tumor size ≥20 mm, and minimal ablative margin<5 mm were risk factors for LTPFS; extrahepatic metastasis, tumor burden>30 mm, and lesion with minimal ablative margin<5 mm were independent risk factors for OS; re-intervention was an independent protective factor for OS.Conclusions:Adequate ablative margin and less tumor burden were beneficial to local tumor control and long-term survival of patients in the RFA treatment; the existence of extrahepatic metastasis was an important risk factor for OS, and re-interventional therapy was beneficial to extend OS.

Modified Brisson technique to treat concealed penis in children / 中华整形外科杂志

Objective@#To investigate the corrective effect of modified Brisson technique to treat concealed penis in children.@*Methods@#From July 2015 to July 2017, 32 patients(3-10 years old, mean 7.2 years old )with concealed penis were treated with the modified Brisson technique in the Children′s Hospital of Zhengzhou University. All the children with concealed penis were treated by the modified Brisson technique, complications, postoperative results and followed-ups were retrospectively analyzed.@*Results@#32 cases of concealed penile children had pain on the postoperative day, 2 cases required patient controlled analgesia device. No infection and dehiscence were noticed after discharge. All the children were followed up for 6 months to 1 year, and the results were satisfactory. Review showed that the penile body was similar to the appearance after circumcision, with obvious extension exposure, normal exposure of penile scrotal Angle and penile pubic Angle, and no obvious retraction. The internal plate of the prepuce in 5 cases was slightly edema, and edema dissolved within 2 months.@*Conclusions@#Modified Brisson is a better method to treat concealed penis in children, which can correct the pathological structure and retain the integrity of penile skin to the greatest extent.

Transarterial chemoembolization by using CalliSpheres drug-eluting beads combined with 8Spheres microsphere for the treatment of advanced hepatocellular carcinoma: study on its short-term efficacy and safety / 介入放射学杂志

Objective To investigate the clinical short-term efficacy and safety of transarterial chemoembolization (TACE) by using CalliSpheres drug-eluting beads combined with blank microspheres (8 Spheres) in treating advanced hepatocellular carcinoma (HCC) . Methods The clinical data of 98 HCC patients, who were treated at authors′ hospital during the period from July 2016 to November 2017, were retrospectively analyzed. The patients were divided into DEB-TACE group (receiving TACE by using CalliSpheres drug-eluting beads combined with 8 Spheres embolic microspheres, n =50) and cTACE group (receiving TACE by using opium poppy ethyl iodized oil together with 8 Spheres embolic microspheres, n =48) . The disease remission rate, control rate and incidence of complications as well as preoperative and 3-day postoperative hepatic function indexes were collected, and the results were compared between the two groups. Results No statistically significant differences in baseline data existed between the two groups (P>0.05) . Both the 3-month and the 6-month disease remission rates in DEB-TACE group were remarkably higher than those in cTACE group (P<0.05), besides, the 6-month disease progression rate and the incidence of elevated serum AFP in DEB-TACE group were lower than those in cTACE group, the short-term efficacy of DEB-TACE group was better than that of cTACE group (P<0.05) . Three days after treatment, the levels of ALT and AST in cTACE group were strikingly higher than those in DEB-TACE group (P<0.05) . No serious adverse reactions occurred in both groups. Conclusion In treating advanced HCC with TACE, the short-term efficacy and safety of CalliSpheres drug-eluting beads combined with 8 Spheres embolic microspheres are superior to those of opium poppy ethyl iodized oil together with 8 Spheres embolic microspheres.

The influence of uterus position on HIFU ablation effect of uterine fibroids / 介入放射学杂志

Objective To compare high-intensity focused ultrasound (HIFU) ablation effect on uterine fibroids between patients with different uterus positions. Methods The clinical data of 372 patients with symptomatic uterine fibroids, who were treated with HIFU at authors' hospital, were retrospectively analyzed. According to the position of the uterus, the patients were divided into anteverted group, mid-position group and retroverted group. Single factor analysis of variance was used to compare the baseline data of fibroids and HIFU ablation results between each other among the three groups. Results Successful HIFU treatment was accomplished in all patients. The average age of the patients was (39±6) years. Single factor analysis of variance indicated that no statistically significant difference in baseline data of fibroids existed between each other among the three groups (P>0.05) . The one cm3 sonication time and energy efficiency factor (EEF) in the anteverted group were significantly smaller than those in the retroverted group, the differences were statistically significant (P =0.007 and P =0.017 respectively) . Conclusion Uterine fibroids within an anteverted uterus are more likely to be ablated by HIFU.

EffectofhighGintensityfocusedultrasoundablationonuterinefibroidswithdifferentsignalintensityonT2WI / 实用放射学杂志

Objective ToinvestigatetheeffectofhighGintensityfocusedultrasound (HIFU)ablationonuterinefibroidswithdifferent withdifferentT2Gweightedimage(T2WI)signalintensity.Methods 357patientswithsymptomaticuterinefibroidstreatedwith HIFU wereretrospectivelyanalyzed.AccordingtoT2WIsignalcharacteristics,fibroidsweredividedinto4groups:hypointensity,isointensity, hyperintensityand mixedintensity.The HIFUtreatmentparametersandablationeffectsofeachgroup werecomparedby O n eG W a y analysisofvariance.Results Theaverageageofthepatientswas(38.5±6.2)y,andtherewerenosignificantdifferencesinbaseline data(P＞0.05).AllpatientssuccessfullycompletedHIFUtreatment,withtheaveragetreatmenttimeof(128.3±60.9)min,theaverage nonGperfusionvolumerate(NPVR)of(74.4±14.7)%,andtheaverageenergyefficiencyfactor(EEF)of(7.1±4.8)j/mm3.Theresults of O n eG W a y analysisofvarianceshowedthattheT2WIhypointensitygrouphadshortersonicationtime,treatmenttimeand1cm3 sonicationtime,smallertreatmentpowerandEEF,largerNPVR,comparingwiththehyperintensitygroup withstatisticallysignificant differences(P＜0.05).Conclusion UterinefibroidswithT2WIhypointensityhavelowwatercontentandhighenergydepositionefficiency, whicharemoresuitableforHIFUablation.

Risk factors of hemorrhagic brain arteriovenous malformation / 中国介入影像与治疗学

Objective To analyze risk factors of hemorrhagic brain arteriovenous malformation (BAVM).Methods Totally 105 BAVM patients confirmed with DSA were collected,and corresponding risk factors were analyzed.Results Univariate analysis suggested that factors were correlated to hemorrhagic presentation including the site and size of the lesions,the type and number of feeding arteries,the number,type and form of draining veins and co-existing arteriovenous fistula.Multiple factors Logistic analysis showed that the location,size,the type and amount of supplying arteries,the number of the draining vein and combined with arteriovenous fistula were the risk factors for hemorrhagic presentation,and drainage vein dilation might be a protective factor.Conclusion Infratentorial and deep cerebral lesions,arteriovenous malformation with diameter ＜3 cm,single draining vein,pure perforating artery,multiple feeding arteries and combined with arteriovenous fistula are risk factors for hemorrhagic BAVM.

Risk factors and interventional therapy of hemorrhage in brain arteriovenous malformation associated with aneurysm / 中国介入影像与治疗学

Objective To explore the risk factors and interventional strategies of bleeding caused by brain arteriovenous malformations associated with aneurysms.Methods Clinical and imaging data of 42 cases of brain arteriovenous malformations associated with aneurysms were collected and analyzed.According to the results of intraoperative angiography and the architectural characteristics of vessels,the therapeutic schedule was formulated.Three months,6 months and 1 year after interventional management,all patients were followed with DSA to observe whether arteriovenous malformations recurred or not.Results The risk factors of hemorrhage caused by brain arteriovenous malformations associated with aneurysms included lesions located in subtentorium,deep vein or mixed venous drainage,Spetzler-Martin grade Ⅲ-Ⅳ,as well as type Ⅰ and type Ⅱ a aneurysms of Redekop classification.Glasgow outcome score of 38 postoperative patients were more than 4 points,visual impaired happened in 1 patient,whereas 2 patients had different degree of limb dysfunction and 1 patient died.DSA follow-up found no recurrence of arteriovenous malformation in 37 patients.Conclusion Understanding the vessel architectural characteristics and substantial hemorrhage risk factors before intervention,therefore formulating individualized therapeutic schedules are benefit to improving safety and efficacy for the treatment of brain arteriovenous malformations associated with aneurysms.

Recent advance in mesenchymal stem cells in treatment of ischemic stroke / 中华神经医学杂志

The treatment of ischemic stroke remains clinically a daunting task as few therapeutic strategies have proven to be effective.So far,a large number of animal experiments and clinical trials have confirmed the unique biological characteristics and therapeutic effect of mesenchymal stem cells on cerebral ischemia.Therapeutic strategies based on transplantation ofmesenchymal stem cells hold great promise for treatment of ischemic stroke.Here,we review mechanisms of mesenchymal stem cells in the treatment ofischemic stroke,transplantation pathways,time windows and tracer methods,providing a good reference to research and apply mesenchymal stem cells in the treatment of stroke.

Study on influence of Toxoplasma tachyzoites on human sperm motility parameters in vitro / 中国血吸虫病防治杂志

Objective To explore the influence of Toxoplasma tachyzoites infection on motility of human spermatozoa in vitro, and explain its possible mechanism. Methods Semen samples obtained from 10 healthy volunteers by masturbation were prepared by the swim-up technique. The samples were then inoculated at 37 ℃ with different concentrations of live Toxoplasma tachyzoites varying among 1?103/ml (Group A), 1?104/ml (Group B), 1?105/ml (Group C), 1?106/ml (Group D), 1?107/ml (Group E) and Group F containing Ham’s F-10 only as the negative control. Motion parameters were analysed by Computer-aided sperm analyzer(CASA) in 0 hour, 1 hour, 2 hours, 4 hours respectively. Modalities of spermatozoa and possible adherence and/or agglutination were observed under the light microscope. Results Distinct adhesion of spermatozoa to Toxoplasma tachyzoites and agglutination were noticed. In all the motion parameters, progressive motility was affected most and dependent upon the incubation time and tachyzoites concentration. Progressive motility showed a significant difference between Group E and the control (P