Objective To explore the role of granzyme B( GzmB) in infiltrated myocardial cells of mice with viral myocarditis( VMC) induced by coxsackievirus B3(CVB3) during the injury of myocardium and further to elucidate the pathogenesis of VMC. Methods Thirty-six BALB/c male mice, 5 to 6 weeks old, were randomly divided into two groups: the normal group, 10, and the model group, 26. On the eleventh day all mice were killed and the hearts were taken out for inspection whether there were pathological changes in the myocardium by macroscopy and microscopy. The expression of GzmB was detected in myocardium with immunohistochemistry and hemi-quantitive reverse transcript-polymerase chain reaction ( RT-PCR). Results GzmB positive cells in the mice myocardial cells infiltrated were detected by immunohistochemistry in the model group,and the number of GzmB positive cells is remarkable positive correlation to myocardial pathological score(r =0. 859, P =0.000) , however, no such cells were discovered in the normal murine hearts. Test of RT-PCR showed markedly high expression of GzmB mRNA in mice myocardium of the model group, however, no expression of GzmB mRNA in the normal group. Conclusion The expression of GzmB in the infiltrating myocardiocytes probably plays some role during the injury of myocardium, which mediated cytotoxicity effect is one of important mechanisms of myocardial injury in viral myocarditis. The expression quantity of GzmB in the infiltrating myocardiocytes is significant positive correlation to the severity of myocardial injury.

Humans ; Phosphorus Compounds ; toxicity ; Poisoning ; therapy

Objective To construct a subtractive cDNA library of genes transactivated by XTP3 protein with suppression subtractive hybridization technique in order to clone genes associated with transactivation. Methods mRNA was isolated from HepG2 cells transfected by pcDNA3.1(-)-XTP3 and pcDNA3.1(-) empty vector respectively, and then cDNA was synthesized. After restriction enzyme Rsa Ⅰ digestion, small sizes cDNAs were obtained. Then tester cDNA was divided into two groups and ligated to the specific adaptor 1 and adaptor 2 respectively. After being hybridized with driver cDNA twice and underwent two times of nested PCR, tester cDNA was subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E.Coli strain JM109. The cDNA was sequenced and analyzed in GenBank with Blast search after PCR. Results The amplified library contained 30 positive clones. Colony PCR showed that 23 clones contained 200-1 000bp inserts. Sequence analysis was performed also. It was found that 20 kinds of known and 2 kinds of novel cDNA sequences may be target genes transactivated by XTP3 protein. Conclusion The subtractive library of genes transactivated by XTP3 protein was constructed successfully. It provides a foundation for the further research on pathogenesis of the viral proteins.

Objective To investigate the correlation between serum soluble epithelial cadherin (sE-cad) and postoperative recurrence and prognosis in patients with advanced gastric cancer.Methods The level of serum sE-cad in 85 patients with advanced gastric cancer (advanced gastric cancer group) was detected by ELISA technique preoperative and postoperative 1 month,and compared with 30 healthy controls(control group).The patients in advanced gastric cancer group were followed up for 3 years,the level of serum sE-cad in recurrent patients and non-recurrent patients was compared.Results The level of serum sE-cad in advanced gastric cancer group preoperative was significantly higher than that in control group [(24.3 ± 14.8) μ g/L vs.(9.4 ± 3.8) μ g/L,P ＜ 0.01].The level of serum sE-cad in advanced gastric cancer group was significantly decreased postoperative 1 month [(12.5 ± 6.4) μ g/L vs.(24.3 ± 14.8) μ g/L,P ＜0.01].The level of serum sE-cad in recurrent patients was significantly higher than that in non-recurrent patients and postoperative 1 month [(20.7 ±9.8)μg/L vs.(12.5 ±6.4),(14.8 ±6.2) μg/L,P＜0.01].Univariate analysis revealed that preoperative high serum sE-cad level was related with tumor size,differentiated degree,lymph node metastasis ratio,depth of tumor invasion (P ＜0.05),but had no relationship with histological type(P＞ 0.05).Elevated preoperative serum sE-cad level negatively affected the postoperative survival rate and recurrence rate.Multivariate Logistic regression anaiysis revealed that preoperative serum sE-cad level was an independent risk factor for postoperative 3 years survival rate in advanced gastric cancer (HR =2.068,P =0.013).Conclusions Preoperative elevated serum sE-cad level is related with pathologic features in patients with advanced gastric cancer,and may be an important prognostic factor.Postoperative monitoring the level of serum sE-cad is useful for evaluating the prognosis and recurrence.

Abstract： Objective　To analyze the species distribution of microorganisms in culture tubes reported positive byBACTEC™ MGIT960 (hereafter referred to as "MGIT960") after species identification using matrix-assisted laser desorption/ionization time of flight massspectrometry (MALDI-TOF MS) technique. Methods　From 2021 to 2022, a total of 2 662 positive tubes reported by the MGIT 960 instrument at Tuberculosis Reference Laboratory of Tianjin Center for Tuberculosis Control were collected. Liquid cultures were independently inoculated to blood plate and neutral L-J medium, and the resulting isolate strains were identified using the MALDI-TOF MS method. According to the MALDI-TOF MS results, the non-repetitive results of the same patient on the same culture medium were analyzed for the composition ratio of strain distribution. For the strains not identified by MALDI-TOF MS, 38 strains were selected for 16S rRNA gene sequencing. Results　A total of 605 isolates were obtained from blood plates, and 501 of those were analyzed. Among them, Mycobacterium accounted for 17.76% (89/501), predominant by Mycobacterium abscess 10.18% (51/501) and Mycobacterium fortuitum 3.19% (16/501). Bacteria other than Mycobacterium accounted for 68.06% (341/501), with the main ones being Nocardia farcinica 15.57% (78/501), Gordonia sputa 9.38% (47/501) and Gordonia bronchialis 7.58% (38/501). There were 71 unidentifiable strains, making up 14.17% (71/501). A total of 2 378 strains were isolated from neutral L-J mediums, 1 748 of which were used in the incoming analysis. Among these, 78.72% (1 376/1 748) were Mycobacterium, 60.53% (1 058/1 748) were Mycobacterium tuberculosis (MTB), 4.69% (82/1 748) were Mycobacterium chimaer intracellulare group and 3.55% (62/1 748) were Mycobacterium Lentiflavum. Bacteria other than Mycobacterium accounted for 17.11% (299/1 748) in neutral L-J medium isolates, with Nocardia farcinica 4.35% (76/1 748), Gordonia sputa 2.69% (47/1 748) and Gordonia bronchialis 2.12% (37/1 748) as the main species. There were 73 strains that couldn't be identified, comprising 4.18% (73/1 748). The 38 strains that not identified by MALDI-TOF MS were all found to be Actinobacteria and Firmicutes by sequencing. Conclusions　A variety of nontuberculous Mycobacterium and bacteria other than Mycobacterium were found in the positive culture tubes reported by the MGIT960 instrument, most of which could be quickly identified by mass spectrometry. Bacteria other than Mycobacterium are mainly Nocardia and Gordonia, which should be paid attention to in differential diagnosis.

Abstract: Objective　To understand the characteristics of mutations associated with resistance among 72 multidrug-resistant tuberculosis (MDR-TB) strains using whole genome sequencing (WGS) and to evaluate the performance of WGS for predicting MDR-TB drug resistance. Methods　The clinical strains isolated from patients who visited the outpatient department of Tianjin Center for Tuberculosis Control from January to September in 2020 were collected. Identification tests using p-nitrobenzoic acid (PNB) medium were performed. Drug susceptibility tests (proportion method) on L-J medium were performed. After excluding duplicate strains, 72 MDR-TB strains were selected for WGS. Data were analyzed by using online databases and the phenotypic drug susceptibility test results were compared with resistance profiles predicted by WGS. Results　All of 72 MDR-TB strains belonged to linage 2, and there was no significant difference in rate of pre-extensive drug-resistant tuberculosis (pre-XDR-TB) between modern type and ancestral type (χ2=0.287, P=0.592). A total of 81 mutation types were found from resistance-related genes for 12 anti-tuberculosis drugs, and the common mutation types in different drug-resistant strains were: streptomycin (SM): rpsL Lys43Arg; isoniazid (INH): katG Ser315Thr; rifampicin (RIF): rpoB Ser450Leu; ethambutol (EMB): embB Met306Val; ofloxacin (OFX), levofloxacin (LFX), moxifloxacin (MFX): gyrA Asp94Gly; kanamycin (KAM), capreomycin (CAP), amikacin (AMK): rrs 1401a>g; para-aminosalicylic acid (PAS): folC Ile43Thr. Nine mutation types were found in 9 prothionamide (PTO)-resistant strains, one type for each strain. The sensitivity and specificity of WGS for predicting resistance to different drugs were SM: 98.15% and 88.89%, INH: 90.28% and -, RIF: 98.62% and -, EMB: 79.49% and75.76%, OFX: 97.30% and 85.71%, KAM: 85.71% and 98.46%, PAS: 27.27% and 95.08%, PTO: 81.82% and 60.66%, CAP: 60.00% and 98.51%, LFX: 97.22% and 83.33%, MFX: 97.30% and 85.71%, AMK:100.00% and 100.00%, respectively. Conclusion　WGS is a rapid and promising method which has high consistency with the phenotypic drug sensitivity test. Therefore, it has good application prospects in predicting drug resistance in MDR-TB.

Objective To explore the impairment of time perception in patients with Parkinson's disease (PD).Method Thirty-two individuals with idiopathic PD were compared with 32 matched healthy controls using a time reproduction task.Result Compared with healthy controls for the 3-second duration reproduction task, PD patients in the setting of a delay by 1 s ( (2.78 ±0.31 ) s and (2.15 ±0.34) s,respectively) and a delay by 5 s ( (2.74 ±0.32) s and (2.08 ±0.37) s, respectively) showed impairment in time perception with a significant under-estimation of the duration ( t = - 7.459, - 7.533, both P ＜0.01 ).When compared with healthy controls for the 5-second duration reproduction task, PD patients in the setting of a delay by 1 s ( (3.99 ±0.39) s and (3.11 ±0.52) s) and 5 s ( (4.05 ±0.40) s and (2.96 ±0.54) s) also showed impairment in time perception with a significant under-estimation of the duration (t =-7.609 and - 9.120, both P ＜ 0.01 ).When compared with healthy controls for the 600-millisecond duration reproduction task, PD patients in the setting of delay by 1 s ((0.91 ±0.18) s and (0.90 ±0.18) s, respectively) and by 5 s ( (0.89 ± 0.16) s and (0.91 ± 0.17 ) s, respectively) did not have impaired time perception, and the difference was not significant ( t = 0.347, P = 0.730; t = - 0.519, P =0.606, respectively).Conclusion Our data indicates that time perception is impaired in PD patients.Impaired time perception is mainly an under-estimation of the time interval in seconds rather than milliseconds.

Objective To provide reference for establishing diagnosis and differential diagnosis methods of rare yeast-like fungal bloodstream infection for clinical microbiology laboratory.Methods Trichosporon asahii (T.asa-hii)and Geotrichum capitatum (G.capitatum)bloodstream infection was diagnosed and differentially diagnosed through clinical data analysis,morphological examination,biochemical reactions,and molecular biology technology. Results Two types of yeast-like fungal bloodstream infections in case 1 and case 2 both occurred in leukemia agranulocytosis phase after chemotherapy,such infections were serious and highly similar.The cultivated colonies on blood agar plates of case 1 and case 2 were performed gram stain and microscopic examination.Hyphae,arthro-spores and microconidia were visible in the former,thickness of hyphae branches and length of arthrospores were different,most presented rectangular and barrel shape;the latter can be seen hyphae with transparent septum bro-ken up into arthrospores,presented rectangular shape,did not produce blastoconidia.Identification with API 20C AUX showed that they were T.asahii and G.capitatum.The PCR product sequences were compared with NCBI, suggesting that T.asahii and G.capitatum were at sexual stage.Conclusion Comprehensive application of a varie-ty of technical methods is helpful for improving the diagnosis accuracy of bloodstream infection with yeast-like fungi, identifying Trichosporon and Geotrichum to the species level may help physicians to understand such rare fungal in-fection,choose antifungal agents rationally,and improve clinical prognosis.

Objective:The aim of the current study was to explore the alterations of TLR4 after traumatic brain injury. Methods: 20 Male ICR rats were randomly divided into four groups(5 rats each group)including control group without brain injury and traumatic brain injury groups(4,24 hours and 7 days).All rats were decapitated at corresponding time point and mid-jejunum samples were taken.The intestinal mucosa structure was detected by histopathological examination.Immunohistochemistry and realtime RNA were used for detection of TLR4 expression in ileum samples.Results: After traumatic brain injury,the histopatholocal alterations of gut mucosa occurred at 4 hours and progressed to a serious state.Compared to control group,TLR4 was significantly up-regulated on the endothelia of microvessels in villous interstitium and lamina propria by 4 h following TBI and maximally expressed at 24 h post-injury.The endothelial TLR4 immunoreactivity in ileum mucosa still remained strong on 7 d post-injury.There was a very low mRNA expression of TLR4 in the gut of control rats and significantly increased at 4 h following TBI(P

Objeetive To study the differences of the biological characteristics and immune regulation function of adipose mesenchymal stem cells (AMSCs)from psoriasis patients and healthy people.Methods AMSCs were isolated and cultured from human psoriatic and healthy adipose tissue,the phenotypes and cell cycle of AMSCs taken from three generation were detected by flow eytometry.Alkaline phosphate enzyme staining and oil red o staining were used respectively to identify their adipogenic and osteogenic capacity.Next,the levels of inflammation antimicrobial proinflammatory factor were detected by PCR and ELISA.Then gene expression profile of AMSCs were screen by gene expression profile chip,as so to bolting the the gene array related with immunology gene.Results There was no significant change in cell morphology,and cell surface markers were expressed high for CD29,CD44,CD73,while lower for CD31,CD45 and HLA-DR.AMSCs of psoriasis patients and healthy people both had the ability of adipogenic and osteogenic differentiation.But the cell cycle showed the third generation AMSCs proliferation rates were slower than that of normal control,as compared with healthy controls,adipogenic differentiation ability was stronger.What'more,the level of inflammatory cytokines in psoriasis group was lower than that in controls such asIL-10,IDO,TGF-β,on the contrary the levels of proinflammatory factor in psoriasis group were higher than that in controls,such as TNF-α,IFN-γ.In addition,gene chip results suggested that psoriasis group AMSCs had obvious expression differences on JAK-STAT pathway with healthy controls.Conclusions Compared with the control,there are significant differences in patients AMSCs proliferation and adipogenic differentiation ability,immune inflammation suppression control ability is weaken,this phenomenon may be associated with JAK-STAT immune pathways related to downgrade.