[Expression of glycolytic genes in immune cells and changes of related immune cells in experimental autoimmune neuritis].

Objective: To investigate the expression of glycolytic genes in immune cells and the changes of related immune cells in experimental autoimmune neuritis (EAN), and deepen the understanding of pathogenesis of EAN. Methods: Twenty-four male C57BL/6 mice (6-8 weeks old, 18-20 g) were divided into four groups according to the random number table method: control group (P0180-199 was replaced by PBS during modeling and mice were sacrificed on the 16th day), EAN mice were sacrificed on the 8th day after the end of modeling (EAN 8 d), EAN mice were sacrificed on the 16th day after the end of modeling (EAN 16 d), and EAN mice received drug intervention and were sacrificed on the 16th day after the end of modeling (2-DG was intraperitoneally injected since the day of the first immunization, 550 mg/kg; EAN 16 d+2-DG), with 6 rats in each group. The clinical symptoms and clinical scores were observed and recorded daily. At the end of the experiment, the mice were sacrificed under chloral hydrate anesthesia, and the serum, spleen, sciatic nerve and other tissues of each group were collected. The degree of inflammatory cell infiltration and demyelination of sciatic nerve were observed by hematoxylin and eosin (HE) staining and luxol fast blue (LFB) staining. Flow cytometry was used to detect the proportion of M1 macrophages, Th17 cells and Tregs cells. The mRNA expression levels of glycolysis-related genes (mTORC1, HIF1α, GLUT1 and LDHA) were detected by RT-PCR. Western blotting was used to detect the level of pan-lysine lactate in macrophages and sciatic nerve tissue. Results: The expression of glycolysis-related genes (mTORC1, HIF1α, GLUT1 and LDHA) in spleen M1 macrophages and sciatic nerve was significantly up-regulated in EAN 16 d group, compared with control, EAN 8 d and EAN 16 d+2-DG groups (all P<0.05). The relative pan-lysine lactate (pankla) expression level of spleen M1 macrophages (1.25±0.02) and sciatic nerve tissue (1.23±0.26) significantly increased in EAN 16 d group, compared with control, EAN 8 d and EAN 16 d+2-DG groups (M1 macrophages: 0.12±0.10, 1.07±0.12 and 0.42±0.07; sciatic nerve: 0.10±0.12, 0.87±0.20 and 0.36±0.05) (all P<0.05). The expression of glycolytic genes in splenic CD4+T cells showed an increasing trend, but there were no statistically significant differences among the groups, and the expression of glycolytic genes did not decrease significantly after 2-DG treatment (all P>0.05). The proportion of spleen M1 macrophages in the control group, EAN 8 d group, EAN 16 d group and EAN 16 d+2-DG group was 4.28±0.13, 7.54±0.25, 13.16±0.33 and 4.13±0.38 respectively, which was significantly higher in the EAN 16 d group (all P<0.05). The proportion of spleen Th17 cells in the four groups was 3.78±0.03, 8.24±0.55, 12.30±1.34 and 4.83±0.01, respectively, which was significantly higher in the EAN 16 d group (all P<0.05). The proportion of spleen Tregs cells in the four groups was 10.01±1.05, 7.54±0.70, 3.82±0.47 and 8.22±1.21, respectively, which was significantly lower in the EAN 16 d group (all P<0.05). Conclusions: The expression of glycolytic genes in splenic macrophages significantly increases during EAN, but not in CD4+T cells. The proportion of M1 macrophages and Th17 cells in spleen gradually increases, while the proportion of Tregs cells gradually decreases.

[Correlation analysis between blood routine-derived inflammatory markers and respiratory function in pneumoconiosis patients].

Objective: To analyze the correlation between blood routine-derived inflammation indicators and respiratory function in patients with pneumoconiosis. Methods: In January 2021, 492 male pneumoconiosis patients hospitalized in Hefei Institute of Occupational Disease Control and Prevention from 2012 to 2020 were randomly selected as the case group, 492 dust exposed non pneumoconiosis workers who underwent occupational health examination at the same time were taken as the control group. The occupational history and clinical examination data of the two groups of subjects were collected, the correlation between blood routine-derived inflammatory indexes and pulmonary function and blood gas analysis was analyzed retrospectively. Results: Compared with the control group, the lymphocyte monocyte ratio (LMR) in the case group was decreased, and the neutrophil lymphocyte ratio (NLR) was increased, and the difference was statistically significant (P<0.05) . There were significant differences in forced vital capacity as a percentage of the predicted value (FVC) , forced expiratory volume in the first second as a percentage of the predicted value (FEV(1)%) , one second rate (FEV(1)/FVC) , partial pressure of oxygen (PaO(2)) , partial pressure of carbon dioxide (PaCO(2)) , and pH among pneumoconiosis patients at different stages (P<0.05) . FVC%, FEV(1)%, FEV(1)/FVC, and PaO(2) decreased with the increase of the stage, the trend test was statistically significant (tau-b=-0.24, -0.34, -0.37, -0.17, P<0.05) , PaCO(2) and pH increased with the increase of the stage, and the trend test was statistically significant (tau-b=0.10, 0.08, P<0.05) . There were statistically significant differences in LYM, LMR, NLR, platelet lymphocyte ratio (PLR) in patients with pneumoconiosis at different stages (P<0.05) , and LYM and LMR decreased with the increase of stage, trend test showed that there was statistically significant (tau-b=-0.11, -0.13, P<0.05) . There were significant differences in FVC%, FEV(1)%, FEV(1)/FVC, PaO(2), pH, LMR, NLR, PLR among patients with different types of pneumoconiosis (P<0.05) . LMR in pneumoconiosis patients was significantly positively correlated with FVC%, FEV(1)%, FEV(1)/FVC and PaO(2) (r(s)=0.342, 0.324, 0.203, 0.207, P<0.05) , NLR was significantly negatively correlated with FVC%, FEV(1)%, FEV(1)/FVC and PaO(2) (r(s)=-0.193, -0.202, -0.164, -0.177, P<0.05) , PLR was significantly negatively correlated with FVC%, FEV(1)%, FEV(1)/FVC and PaO(2) (r(s)=-0.194, -0.193, -0.106, -0.113, P<0.05) . Multiple linear regression analysis showed that LMR in pneumoconiosis patients was positively related with FVC%, FEV(1)% and PaO(2) (P<0.05) . Conclusion: LMR in patients with pneumoconiosis has a certain correlation with lung function and blood gas analysis, LMR is expected to become a sensitive indicator for evaluating pneumoconiosis.

Forensic Analysis of 43 Medical Disputes Caused by Death after Cardiac Surgery.

ABSTRACT: Objective To explore the causes and characteristics of medical disputes caused by death after cardiac surgery and to analyze the pathological changes after cardiac surgery and the key points of forensic anatomy, thus to provide pathological evidence for clinical diagnosis and treatment of cardiac surgery and judicial appraisal as well as reference for the prevention of medical disputes in such cases. Methods Forensic pathological cases of medical disputes caused by death after cardiac surgery which were accepted by the Center for Medicolegal Expertise of Sun Yat-Sen University from 2013 to 2018 were analyzed retrospectively from aspects such as causes of death, pathological diagnosis, surgery condition, medical misconduct, and so on. Results The causes of death after cardiac surgery of 43 patients were abnormal operation, low cardiac output syndrome, postoperative infection, postoperative thrombosis, and other diseases. Among the 43 cases, there were 18 cases without medical fault while 25 cases had medical fault. Conclusion The medical disputes caused by death after cardiac surgery are closely related to the operative technique and postoperative complications. The causes of medical faults include defects in diagnosis and treatment technique, as well as unfulfillment of duty of care.

[Analysis of related risk factors in the occurrence of mercury-toxic nephrotic syndrome].

Objective: To analyse the clinical characteristics of mercury-toxic nephrotic syndrome and to find the risk factors. Methods: A total of 126 patients with mercury-toxic admitted to Beijing Chaoyang Hospital from January 2017 to January 2020 were selected as the research objectives. General data such as age, gender, mercury exposure history of all patients and laboratory data such as urine mercury and urine creatinine were collected. The clinical manifestations and pathological types of patients with mercury-toxic nephrotic syndrome were analyzed, and the risk factors of mercury-toxic nephrotic syndrome were found by univariate and multivariate logistic regression analysis. Results: Among the 24 patients with mercury-toxic nephrotic syndrome, 1 was poisoned by respiratory tract, 6 were poisoned by digestive tract, and 17 were poisoned by skin absorption. In addition to the manifestations of kidney injury, some of the patients also showed neurological symptoms such as dizziness, fatigue, hand tremor, and limb pain. The main pathological types were membranous nephropathy (9 cases) and micropathological nephropathy (10 cases) . The results of multivariate logistic regression analysis showed that the digestive tract and skin absorption pathways of mercury were risk factors for mercury-toxic nephrotic syndrome (OR=21.099, 23.840, P<0.05) . Conclusion: The absorption pathway of mercury has an important influence on the pathogenesis of mercury-toxic nephrotic syndrome, especially in patients with mercury poisoning absorbed by the skin and digestive tract, the risk of complicating nephrotic syndrome is higher.

The single nucleotide polymorphism rs11643718 in SLC12A3 is associated with the development of diabetic kidney disease in Chinese people with type 2 diabetes.

AIMS: To examine the association between 24 literature-based single nucleotide polymorphisms and diabetic kidney disease in Chinese people with type 2 diabetes. METHODS AND RESULTS: Twenty-four candidate diabetic kidney disease-susceptible single nucleotide polymorphisms were genotyped in 208 participants with type 2 diabetes and diabetic kidney disease and 200 participants with type 2 diabetes without diabetic kidney disease (case and control groups, respectively), together with 206 healthy participants using MassARRAY. Rs11643718 in the SLC12A3 gene was associated with diabetic kidney disease in the recessive model after adjusting for confounding factors, such as age and gender (adjusted odds ratio 2.056, 95% CI 1.120-3.776; P = 0.020). Meta-analyses further confirmed the association (P = 0.002). In addition, participants with the GG genotype had worse renal function and more albuminuria than those with the AA+AG genotype (P < 0.05). Renal section immunohistochemistry was conducted in participants with type 2 diabetes, diabetic kidney disease and AA+AG or GG genotypes and in participants with glomerular minor lesions. Together with data from the Nephroseq database, it was shown that the abundance of SLC12A3 was reduced in patients with the GG genotype, while elevated expression of SLC12A3 was associated with better renal function. In addition, rs10951509 and rs1345365 in ELMO1, which were determined to be in high linkage disequilibrium by SHEsis software, were also associated with diabetic kidney disease (adjusted P = 0.010 and 0.015, respectively). CONCLUSIONS: The G allele and GG genotype of SLC12A3 rs11643718 are associated with the development of diabetic kidney disease in a Chinese population with type 2 diabetes.

FBLN3 inhibited the invasion and metastasis of colorectal cancer through the AKT/mTOR pathway.

Fibulin-3(FBLN3) levels are different in different types of cancers. We found that fibulin-3 was downregulated in colorectal (CRC) cells, particularly in the SW480 cell line. By comparison, transfecting SW480 cells with a lentivirus overexpressing fibulin-3 RNA could inhibit proliferation, induce G1/S arrest, and promote cell apoptosis. Fibulin-3 overexpression further suppressed the invasion and metastasis of CRC. These effects were regulated through the AKT/mTOR signaling pathway.

[Analysis of differences in epidemiology and clinical features of Guillain-Barré syndrome between rural and urban areas of southern China].

Objective: To explore the differences in epidemiology and clinical features of Guillain- Barré syndrome (GBS) between rural and urban areas of southern China. Methods: The clinical data of 759 hospitalized GBS patients from 31 hospitals of 13 provinces/cities in southern China, between January 1st, 2013 and September 30th, 2016, were collected and analyzed retrospectively. Results: The risk of GBS was higher for males than females in rural and urban areas and the median age was 49 and 48 years, respectively. Seasonal clustering in winter and spring was noted in both rural and urban areas, and the seasonal trend was more markedly in rural areas, but the differences showed no statistical significance. There were 70.37% of patients in rural areas and 73.69% in urban areas who had antecedent respiratory infection. The median time from onset to nadir was 7 days, and Hughes Disability Scale at admission, nadir and discharge were (2.95±1.10 vs 2.84±1.15), (3.25±1.11 vs 3.14±1.21), (2.02±1.24 vs 2.00±1.31) in rural and urban areas respectively. Albuminocytologic dissociation was present in 84.34% of patients in rural areas and 84.62% of cases in urban areas. There were 8.65% and 10.94% of cases in rural and urban areas who required mechanical ventilation during hospitalization, respectively. Demyelinating GBS accounted for 53.29% and 48.77%, respectively, in patients with findings of nerve conduction studies available in rural and urban areas. Conclusions: GBS in rural areas of southern China showed male predominance and a peak of spring and winter occurrence, with respiratory infection as the predominated preceding events and demyelinating GBS being main clinical subtype. Winter and spring showed a higher incidence of GBS in rural and urban areas. There were no significant differences of sex, age, preceding events, season trend, progression of disease, clinical subtypes and cerebrospinal fluid investigations in GBS patients between rural and urban areas.

Strategy of the Causes of Death of Dependents.

ABSTRACT: Objective To discuss the methods and strategies to identify the causes of dependents' deaths, as well as provide the experiences that can be used for reference and scientific basis for the forensic identification of the potentially growing deaths of the same kind in the future. Methods The 13 cases concerning death of dependents accepted by Sun Yat-sen University Forensic Center were collected, and the basic information of the dependents were statistically described. The nutritional status, environmental condition and medical care condition were evaluated according to dietary energy, living space, environment and medical treatment condition. Results Among the 13 dependents, there were 11 males and 2 females, with the oldest 74 and the youngest 9 and dwelling time was from 0.4 to 5.6 years. Forensic pathological examination showed that 13 dependents had infectious diseases and 11 were severely dystrophic. There were no fatal mechanical injuries or poisoning in dependents. Molecular pathological screening of 4 cases revealed no pathogenic variants of sudden death susceptible genes. The poor status of the diet, nutrition, living environment and medical care of these dependents were discovered. The direct cause of death of all 13 dependents was identified to be disease. The lack of nutrition, poor living environment and lack of medical care were thought to play a dominant role in causing the deaths of 12 dependants. Conclusion The death identification should follow the judicial procedure. In identification of the causes of death and analysis of the proportion of the affecting factors resulting in death, all factors, including nutrition,environment, medical care, injury and diseases, need to be considered.

Parametric investigations on the saturation intensity of Coumarin 102 for stimulated emission depletion application.

Stimulated emission depletion (STED) microscopy performed using continuous-wave (CW) lasers has been investigated and developed by Willig et al. (Nature Methods, 2007, 4(11):915) for nearly a decade. Kuang et al. (Review of Scientific Instruments, 2010, 81:053709) developed the CW STED microscopy technique with 405 nm excitation and 532 nm depletion beams. In their research, Coumarin 102 dye was adopted and was found to be depletable. In this study, a parametric investigation of the depletion of Coumarin 102 dye is carried out experimentally. The influence of the excitation and depletion beam intensities and dye concentrations on the depletion efficiency are studied in detail. The results indicate the following: (1) The highest depletion occurs for the 100 µM Coumarin 102 solution, with a 1.4 µW excitation beam and a 115.3 mW depletion beam. (2) The minimum saturation intensity (Is) of STED, that is 13 MW cm-2 , is observed when the Coumarin 102 solution concentration is 10 µM. (3) Is values calculated directly from the depletion power derived with the cross-sectional area due to the full-width-at-half-maximum (FWHM) of the depletion beam show poor accuracy, where Is may be overestimated. Thus, a correction factor for the cross-sectional area is proposed. We also find that Is is not exactly constant for a fixed excitation beam power and dye concentration. This trend indicates that the conventional suppression function η(x)=e- ln (2)ISTED(x)/Is derived from picosecond STED may cause errors in evaluating the depletion process in CW STED microscopy.

[HBXIP expression in gastric adenocarcinoma predicts poor prognosis].

Objective: To investigate the localization of HBXIP protein over-expression in gastric adenocarcinoma, and its prognostic significance. Methods: HBXIP localization was detected by immunofluorescence in AGS gastric cancer cell line, and by immunohistochemical staining in 97 gastric adenocarcinomas, 41 adjacent non-tumor tissues and 13 gastric adenoma tissues. Correlation between HBXIP expression and clinicopathological features of gastric cancer patients was evaluated by Chi-square and Fisher's exact tests. Overall survival rates were calculated using Kaplan-Meier method. Results: HBXIP was mainly expressed in the cytoplasm of gastric cancer. The positive and strongly positive expression rates of HBXIP protein in gastric cancers were 68.0% (66/97) and 49.5% (48/97) respectively, and were significantly higher than those in adjacent non-tumor tissues(48.8%, 20/41; 36.6%, 15/41) or gastric adenomas(2/13, 1/13; all P<0.05). HBXIP expression correlated significantly with tumor differentiation and lymph node status (P=0.007; 0.041). Kaplan-Meier survival analysis showed that the overall survival rate was significantly lower in gastric cancer patients with high HBXIP expression (P=0.015). Conclusions: HBXIP expression in gastric cancer is mainly expressed in cytoplasmic, and the expression level is closely related to the prognosis. HBXIP expression status may potentially be used as an important prognostic indicator for gastric cancer.

[Research Progress of the Correlation between Caveolin and Unexpected Sudden Cardiac Death].

Due to the negative autopsy and without cardiac structural abnormalities, unexpected sudden cardiac death （USCD） is always a tough issue for forensic pathological expertise. USCD may be associated with parts of fatal arrhythmic diseases. These arrhythmic diseases may be caused by disorders of cardiac ion channels or channel-related proteins. Caveolin can combine with multiple myocardial ion channel proteins through its scaffolding regions and plays an important role in maintaining the depolarization and repolarization of cardiac action potential. When the structure and function of caveolin are affected by gene mutations or abnormal protein expression, the functions of the regulated ion channels are correspondingly impaired, which leads to the occurrence of multiple channelopathies, arrhythmia or even sudden cardiac death. It is important to study the effects of caveolin on the functions of ion channels for exploring the mechanisms of malignant arrhythmia and sudden cardiac death.

[Correlation between Genetic Variants and Polymorphism of Caveolin and Sudden Unexplained Death].

OBJECTIVES: To explore the genetic variation sites of caveolin （CAV） and their correlation with sudden unexplained death （SUD）. METHODS: The blood samples were collected from SUD group （71 cases）, coronary artery disease （CAD） group （62 cases） and control group （60 cases）, respectively. The genome DNA were extracted and sequencing was performed directly by amplifying gene coding region and exon-intron splicing region of CAV1 and CAV3 using PCR. The type of heritable variation of CVA was confirmed and statistical analysis was performed. RESULTS: A total of 4 variation sites that maybe significative were identified in SUD group, and two were newfound which were CAV1： c.45C>T （T15T） and CAV1：c.512G>A （R171H）, and two were SNP loci which were CAV1：c.246C>T （rs35242077） and CAV3：c.99C>T （rs1008642） and had significant difference （P<0.05） in allele and genotype frequencies between SUD and control groups. Forementioned variation sites were not found in CAD group. CONCLUSIONS: The variants of CAV1 and CAV3 may be correlated with a part of SUD group.

Diversity of endophytic fungi of Myricaria laxiflora grown under pre- and post-flooding conditions.

Myricaria laxiflora is distributed along the riverbanks of the Yangtze River valley. The Three Gorges Dam has dramatically changed the habitat of M. laxiflora, which has evolved to develop increased resistance to flooding stress. In order to elucidate the relationship between plant endophytic fungi and flooding stress, we isolated and taxonomically characterized the endophytic fungi of M. laxiflora. One hundred and sixty-three fungi were isolated from healthy stems, leaves and roots of M. laxiflora grown under pre- and post-flooding conditions. Culture and isolation were carried out under aerobic and anaerobic conditions. Based on internal transcribed spacer sequence analysis and morphological characteristics, the isolates exhibited abundant biodiversity; they were classified into 5 subphyla, 7 classes, 12 orders, 17 families, and 26 genera. Dominant endophytes varied between pre- and post-flooding plants, among different plant tissues, and between aerobic and anaerobic culture conditions. Aspergillus and Alternaria accounted for more than 55% of all isolates. Although the number of isolates from post-flooding plants was greater, endophytes from pre-flooding plants were more diverse and abundant. Endophytes were distributed preferentially in particular tissues; this affinity was constrained by both the host habitat and the oxygen availability of the host.

Genome-wide identification and expression profiling of the fatty acid desaturase gene family in the silkworm, Bombyx mori.

Fatty acid desaturases exist in all living organisms and play important roles in many different biologic processes, such as fatty acid metabolism, lipid biosynthetic processes, and pheromone biosynthetic processes. Using the available silkworm genome sequence, we identified 14 candidate fatty acid desaturase genes. Eleven genes contain 3 conserved histidine cluster motifs and 4 transmembrane domains, but their N-terminal residues exhibit obvious diversity. Phylogenetic analysis revealed that there are 6 groups; Bmdesat1 and Bmdesat5-8 were clustered into group 2, which is involved in Δ11 desaturation activity, and Bmdesat3-4 were grouped in group 1, which is involved in Δ9 desaturation activity. Twelve of the 14 genes have expressed sequence tag evidence. Microarray data and reverse transcription polymerase chain reaction analysis demonstrated that Bmdesat3-4 and Bmdesat10 were expressed from the larval to moth stages and in multiple tissues on day 3 of 5th instar larvae. Bmdesat9, Bmdesat11, and Bmdesat14 were expressed during the pupal and late-embryonic stage, suggesting that they may take part in fatty acid metabolism to provide energy. These results provide some insights into the functions of individual fatty acid desaturases in silkworm.

Clinical Feasibility of Using Single-isocentre Non-coplanar Volumetric Modulated Arc Therapy Combined with Non-coplanar Cone Beam Computed Tomography in Hypofractionated Stereotactic Radiotherapy for Five or Fewer Multiple Intracranial Metastases.

AIMS: To evaluate the clinical feasibility of single-isocentre non-coplanar volumetric modulated arc therapy (NC-VMAT) with non-coplanar cone beam computed tomography (NC-CBCT) in hypofractionated stereotactic radiotherapy (HSRT) for five or fewer multiple brain metastases. MATERIALS AND METHODS: Ten patients with multiple brain metastases who underwent single-isocentre NC-VMAT HSRT with limited couch rotations (within ±45°) and NC-CBCT with a limited scanning range (150-200°) were included in the current analysis. Conventional single-isocentre coplanar VMAT (C-VMAT) plans were generated and compared with NC-VMAT plans. The intracranial response and toxicities of single-isocentre NC-VMAT HSRT were also evaluated. RESULTS: Compared with C-VMAT, NC-VMAT generated better target conformity (P < 0.05), a lower gradient index (P < 0.05) and better normal brain tissue sparing, especially for volume ≥12 Gy, with a median reduction of 12.65 cm3. For 45° couch rotation, NC-CBCT produced sufficient image quality to differentiate bony anatomy, even with a 150° scanning range, which could be successfully used for patient set-up correction. After NC-CBCT, 57.1% of the measured non-coplanar set-up errors exceeded the threshold value. The median gamma passing rate of NC-VMAT was higher than that of C-VMAT plans (P < 0.05). The non-coplanar beam of NC-VMAT with NC-CBCT corrections exhibited superior gamma passing rate to that without NC-CBCT corrections. The intracranial objective response rate and disease control rate for all patients were 80% (8/10) and 100% (10/10), respectively, and the most common toxicities were headache (20%) and dizziness (20%). CONCLUSION: NC-VMAT with limited couch rotation (within ±45°) combined with NC-CBCT with a limited scanning range (150-200°) markedly improves the plan quality and set-up accuracy in single-isocentre multiple-target HSRT.

Can Optical Surface Imaging Replace Non-coplanar Cone-beam Computed Tomography for Non-coplanar Set-up Verification in Single-isocentre Non-coplanar Stereotactic Radiosurgery and Hypofractionated Stereotactic Radiotherapy for Single and Multiple Brain Metastases?

AIMS: To conduct a direct comparison regarding the non-coplanar positioning accuracy between the optical surface imaging system Catalyst HDTM and non-coplanar cone-beam computed tomography (NC-CBCT) in intracranial single-isocentre non-coplanar stereotactic radiosurgery (SRS) and hypofractionated stereotactic radiotherapy (HSRT). MATERIALS AND METHODS: Twenty patients with between one and five brain metastases who underwent single-isocentre non-coplanar volumetric modulated arc therapy (NC-VMAT) SRS or HSRT were enrolled in this study. For each non-zero couch angle, both Catalyst HDTM and NC-CBCT were used for set-up verification prior to beam delivery. The set-up error reported by Catalyst HDTM was compared with the set-up error derived from NC-CBCT, which was defined as the gold standard. Additionally, the dose delivery accuracy of each non-coplanar field after using Catalyst HDTM and NC-CBCT for set-up correction was measured with SRS MapCHECKTM. RESULTS: The median set-up error differences (absolute values) between the two positioning methods were 0.30 mm, 0.40 mm, 0.50 mm, 0.15°, 0.10° and 0.10° in the vertical, longitudinal, lateral, yaw, pitch and roll directions, respectively. The largest absolute set-up error differences regarding translation and rotation were 1.5 mm and 1.1°, which occurred in the longitudinal and yaw directions, respectively. Only 35.71% of the pairs of measurements were within the tolerance of 0.5 mm and 0.5° simultaneously. In addition, the non-coplanar field with NC-CBCT correction yielded a higher gamma passing rate than that with Catalyst HDTM correction (P < 0.05), especially for evaluation criteria of 1%/1 mm with a median increase of 12.8%. CONCLUSIONS: Catalyst HDTM may not replace NC-CBCT for non-coplanar set-up corrections in single-isocentre NC-VMAT SRS and HSRT for single and multiple brain metastases. The potential role of Catalyst HDTM in intracranial SRS/HSRT needs to be further studied in the future.

[Analysis of clinical features and poor prognostic factors of acute hematogenous osteomyelitis in children].

Objective: To analyze the clinical characteristics, pathogenic bacteria, complications and risk factors of prognosis of acute hematogenous osteomyelitis in children. Methods: The clinical manifestations, laboratorg tests, etiological charateristics and clinical data of 107 patients with acute hematogenous osteomyelitis admitted to Beijing Children's Hospital from January 2017 to December 2020 were retrospectively analyzed. According to the drug sensitivity results of Staphylococcus aureus, the group was divided into methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible Staphylococcus aureus (MSSA) group; according to the presence or absence of complications, the group was divided into the group with and without complications; according to the prognosis of the follow-up children, the group was divided into good prognosis and poor prognosis. The χ2 test or Mann-Whitney U test used for comparison between groups, and Logistic regression was used to analyze the risk factors for complications and prognosis. Results: Of the 107 patients, 62 were males and 45 were females. The age of presentation was 5.6 (1.7, 10.0) years, including 5 patients (4.7%) age from >28 days to 3 months, 46 patients (43.0%) age from >3 months to 5 years, 43 patients (40.2%)>5-12 years of age, and 13 patients (12.1%)>12-18 years of age. The first symptoms were acute fever in 35 patients (32.7%), limb pain in 24 patients (22.4%), and fever with limb pain in 23 patients (21.5%). Pathogen culture was positive in 75 patients (70.1%), Streptococcus pyogenes, Salmonella enterica and Escherichia coli in 1 case (1.4%) each, and Staphylococcus aureus in 72 cases (96.0%), among them, 47 cases were MSSA, 22 cases were MRSA, and 3 cases had positive reports of Staphylococcus aureus from other hospitals without drug-sensitive tests. The proportion of infected children living in rural areas and receiving surgical treatment was higher in the MRSA group than in the MSSA group (14 cases (63.6%) vs. 18 cases (38.3%) and 21 cases (95.5%) vs. 33 cases (70.2%), χ2=3.87, 4.23, both P<0.05). Sixty-five children had no complications while 42 children (39.3%) suffered from complications. Common complications consisted of 19 cases (17.8%) of sepsis, 17 cases (15.9%) of septic arthritis, and 12 cases (11.2%) of venous thrombosis. The group with complications showed higher mental changes, decreased appetite and (or) weakness, positive pathogenic cultures, and time from admission to surgery than the group without complications (18 cases (42.9%) vs. 9 cases (13.8%), 20 cases (47.6%) vs. 12 cases (18.5%), 34 cases (81.0%) vs. 41 cases (63.1%), 3.5 (2.0, 6.0) vs. 2.0 (1.0, 4.0) d,χ2=11.38, 10.35, 3.89, Z=2.21, all P<0.05). The poor prognosis group had more comorbidities, combined local complications, and positive aureus than the good prognosis group (10/15 vs. 34.9% (30/86), 7/15 vs. 17.4% (15/86), 14/15 vs. 61.6% (53/86), χ2=5.39, 6.40, 4.42, all P<0.05). Multifactorial Logistic regression analysis showed that acute phase C-reactive protein (CRP) was both an independent risk factor for complications (OR=1.01, 95%CI 1.01-1.02) and an independent risk factor for poor prognosis (OR=1.01, 95%CI 1.00-1.02). Conclusions: The first symptoms of acute hematogenous osteomyelitis are acute fever, limb pain, and fever with limb pain are most common. Staphylococcus aureus is the most common pathogenic organism. Those with loss of appetite and (or) weakness, mental changes, positive pathogenic cultures, and longer time between admission and surgery are prone to complications. Those with complications, combined local complications, and positive for Staphylococcus aureus had a poor prognosis. Elevated CRP is an independent risk factor not only for complications but for poor prognosis as well.

The structural parameters for antimicrobial activity, human epithelial cell cytotoxicity and killing mechanism of synthetic monomer and dimer analogues derived from hBD3 C-terminal region.

Understanding the molecular mechanisms of antimicrobial peptide-membrane interactions is crucial in predicting the design of useful synthetic antimicrobial peptide analogues. Defensins are small (3-5 kDa) cysteine-rich cationic proteins which constitute the front line of host innate immunity. In this study, a series of eight 10 AA C-terminal analogues of hBD3 [sequence: RGRKXXRRKK, X = W, F, Y, V, L, I, H, C(Acm); net charge = +7, coded as W2, F2, Y2, V2, L2, I2, H2, and C2] and covalent V2-dimer [(RGRKVVRR)(2)KK] (18 AA, net charge = +11) were synthesized using solid phase peptide synthesis (SPPS) in Fmoc chemistry. Wild-type hBD3 was used as a control in all analyses. W2, V2, and especially Y2 showed high activity selectively against Gram-negative bacteria Pseudomonas aeruginosa in the concentration range of 4.3-9.7 microM. The covalent dimeric form of V2-monomer, V2-dimer, showed increased antibacterial killing compared to the monomeric form, V2-monomer. Cytotoxicity assays on a human conjunctival epithelial cell line (IOBA-NHC cells) showed that no change in viable cell number 24 h after constant exposure to all the eight peptide analogues even at concentrations up to 200 microg/ml. Fluorescence correlation spectroscopy (FCS) was used to study the interaction of these peptides against POPC vesicles (neutral; mammalian cell membrane mimic) and POPG vesicles (negatively charged; bacterial cell membrane mimic). Using FCS, significant aggregation and some leakage of Rhodamine dye were observed with POPG with Y2, W2 and V2 at the concentration of 5-10 mmicroM and no significant aggregation or disruption of vesicles was observed for all peptide analogues tested against POPC. V2-dimer induced more leakage and aggregation than the monomeric form. Overall, V2-dimer is the most effective antimicrobial peptide, with aggregation of POPG vesicles observed at concentrations as low as 1 microM. The concentration of 5-10 microM for Y2 from FCS correlated with the concentration of 5 microM (6.25 microg/ml), at which Y2 showed a cooperative increase in the activity. This suggests a structural transition of Y2 in the 2.5-5 microM concentration range resulting in the correlated increased antimicrobial activity. These results and the FCS together with previous NMR and molecular dynamics (MD) suggested that the charge density-based binding affinity, stable covalent dimerization, the ability to dimerize or even oligomerize and adopt a well-defined structure are important physicochemical properties distinguishing more effective cationic antimicrobial peptides.

Single nucleotide polymorphism rs2229634 in the ITPR3 gene is associated with the risk of developing coronary artery aneurysm in children with Kawasaki disease.

Kawasaki disease (KD) is the most common form of pediatric vasculitis. Though its etiology is unknown, researchers have suggested that it is related to genetics. The inositol 1,4,5-triphosphate receptor type 3 (ITPR3) gene has a strong association with the development of type 1 diabetes and, plays a critical role in the development of autoimmune diseases such as systemic lupus erythematosus, rheumatoid arthritis, and Graves' disease. The aim of study is to examine the association of ITPR3 polymorphisms with KD risk in Taiwanese children. This study evaluates the single nucleotide polymorphisms (SNP) rs2229634 in the ITPR3 gene with KD in a case-control study involving 93 KD patients and 680 healthy, gender- and age-matched controls. The frequency of the rs2229634 T/T genotype was significantly higher in KD patients with coronary artery aneurysm (CAA) than in patients without CAA [odds ratio (OR) = 2.56, 95% confidence interval (95% CI) = 1.35-4.88, P = 0.004]. In addition, KD patients with the T/T genotype elevated mean serum levels of C-reactive protein compared with patients with the C/C or C/T genotype (12.2 mg dL(-1) vs. 8.5 mg dL(-1) , P = 0.036). In conclusion, the results of this study suggest that the rs2229634 SNP in the ITPR3 gene is associated with the risk of CAA formation in Taiwanese KD patients.