This study aims to investigate the function of lmo2363 gene in stress resistance of Liste-ria monocytogenes strain LM83-1.In this study,the lmo2363 gene deletion strain and complement-ation strain of Listeria monocytogenes were constructed using overlapping extended PCR and ho-mologous recombination techniques,and the growth ability,stress survival rate and biofilm forma-tion ability of wild,deletion strain and complementation strain were compared under different stress environments.lmo2363 gene deletion strain and complementation strain of Listeria monocy-togenes were successfully constructed in this experiment.The growth curves showed that the growth capacity of the deletion strain was weaker than the wild strain LM83-1 under 4 ℃,7％NaCl,10％NaCl,3.5％ethanol,4.0％ethanol and pH5 stress(P＜0.001).The results of stress survival test showed that the survival rate of the deletion strain was significantly lower than the wild strain after 1 h treatment with pH3 and 10 mmol/L H2 O2 stress(P＜0.010).The biofilm forming ability of the deletion strain was decreased compared with that of the wild strain(P＜0.050).This study confirmed that lmo2363 gene mediated the adaptation of LM to low temperature,high osmotic pressure,ethanol and acid stress environment and affected the formation of LM bio-film.This study laid a foundation for further exploring the function of lmo2363 gene in the stress resistance process of Listeria monocytogenes.

Objective:To investigate clinical manifestations, pathological features and diagnosis of eczematoid clear cell acanthoma of the nipple/areola.Methods:The clinical manifestations, histopathological features, special staining results and immunohistochemical features of a case of eczemtoid clear cell acanthoma of the nipple/areola firstly reported in China were analyzed, and compared with those of similar cases in foreign literature.Results:The female patient presented with recurrent pruritic rashes on the left nipple and areola for over 2 years. Skin examination showed hypertrophic skin on the left nipple and areola, and scattered erythema, hypopigmented macules and hyperpigmented macules on the areola, which were covered with a few crusts and scales. Histopathological examination of the skin lesions showed focal epidermal crusts and scales, focal parakeratosis, extended and fused rete ridges, thickened spinous layer, focal spongiosis, clear cell clumps in the spinous cell layer, telangiectasia in the superficial dermis, with infiltration of a few eosinophils and neutrophils. Periodic acid-Schiff staining showed positive results, and immunohistochemical staining revealed positive reaction for epithelial membrane antigen. Topical treatment with triamcinolone acetonide and econazole cream was effective, and topical application of 3% boric acid solution could alleviate exudation. During the 6-month follow-up, the patient experienced intermittent recurrence twice, and responded well to the above treatment.Conclusions:Eczematoid clear cell acanthoma of the nipple/areola has unique clinical and pathological features, revealing that it′s a new subtype of clear cell acanthoma. Pathological examination is the gold standard for its diagnosis.

ObjectiveTo investigate the value of remote consultation of heart sound acquisition in screening and referral of neonates with congenital heart diseases （CHD） in primary hospitals. MethodsA total of 4 030 neonates with non-critical diseases were selected. They were born in Shanghai Pudong New Area Maternal and Child Health Hospital from November 5， 2019 to March 31， 2021. After birth， routine cardiac auscultation was performed and remote consultation of heart sound collection were performed at the same time in combination with percutaneous oxygen saturation measurement to screen CHD. The children with any positive screening index were advised to verify the diagnosis by cardiac ultrasound examination in Shanghai Children's Medical Center. The diagnostic value of different screening methods was compared. ResultsA total of 110 cases were detected positive by routine screening. Among them， 16 cases were lost to follow-up， and 46 cases were confirmed by cardiac ultrasound examination， with a positive diagnosis rate of 48.94% （46/94）. A total of 51 cases were detected positive by routine screening and remote consultation of heart sound collection simultaneously. Among them， 42 cases were confirmed by cardiac ultrasound examination， with a positive diagnosis rate of 82.35% （42/51）. The difference between the two positive diagnosis rates was statistically significant （P<0.001）. ConclusionRemote consultation of heart sound acquisition on the basis of routine neonatal CHD screening can effectively improve the positive diagnosis rate of CHD screening in primary hospitals， and reduce unnecessary referrals. This method is simple and feasible. It has practical value in primary hospitals that lack professional technicians for the diagnosis and treatment of CHD.

Objective:To dynamically observe the effect of N-acetylserotonin (NAS) on the expression of tumor necrosis factor-α (TNF-α) protein in retina of retinal ischemia reperfusion injury (RIRI) rats, and to explore the mechanism.Methods:By using random number table method, 90 healthy male Sprague-Dawley rats were divided into sham operation group ( n=10), RIRI group ( n=40), and NAS group ( n=40). The right eye was as the experimental eye. In the RIRI group and NAS group, the anterior chamber high intraocular pressure method was used to establish the RIRI model. In the NAS group, 10 mg/kg NAS was injected intraperitoneally before modeling and 30 minutes after modeling. At 6, 12, 24, 72 h after modeling, hematoxylin-eosin staining was used to observe the pathological changes of the retina, and the retinal ganglion cells (RGC) were counted. Each group was detected by immunohistochemical staining and Western blot about the relative expression of TNF-α, nuclear factor E2-related factor 2 (Nrf2), and heme oxygenase-1 (HO-1) protein in the rat retina. Oneway analysis of variance was used for differences between groups. The general linear regression method was used to analyze the correlation between the relative expression changes of TNF-α protein and the changes of Nrf2 and HO-1 protein expression after NAS intervention. Results:Optical microscope observation revealed that the retinal edema of rats in the RIRI group was observed at 6, 12, and 24 h after modeling; the thickness of the retina in the NAS group was significantly thinner than that in the RIRI group, and the difference was statistically significant ( F=9.645, 477.150, 2.432; P<0.01). At 6, 12, 24, and 72 h after modeling, the retinal RGC counts in the NAS group were significantly higher than those in the RIRI group, and the difference was statistically significant ( F=12.225, 12.848, 117.655, 306.394; P<0.05). The results of immunohistochemical staining and Western blot showed that 6 h after modeling, the relative expression of TNF-α protein in the retina of the RIRI group increased significantly compared with that in the sham operation group, reaching a higher level at 12 h, and decreased at 24 and 72 h. But all were significantly higher than the sham operation group, the difference was statistically significant (immunohistochemical staining: F=105.893, 1 356.076, 434.026, 337.351; P<0.01; Western blot: F=92.906, 534.948, 327.600, 385.324; P<0.01). At different time points after modeling, the relative expression of TNF-α protein in the retina of the NAS group was significantly lower than that of the RIRI group (immunohistochemical staining: F=15.408, 570.482, 21.070, 13.767; P<0.05; Western blot: F=12.618, 115.735, 13.176, 111.108; P<0.05), but still higher than the sham operation group (immunohistochemical staining: F=40.709, 151.032, 156.321, 216.035; P<0.01; Western blot: F=33.943, 79.729, 74.057, 64.488; P<0.01), the difference was statistically significant; 12 h after modeling, Nrf2 in the retina of the NAS group (immunohistochemical staining: F=51.122, P<0.05; Western blot: F=33.972, P<0.05), HO-1 (immunohistochemical staining: F=30.750, P<0.05; Western blot: F=18.283, P<0.05) protein relative expression was significantly higher than that of RIRI group, and the differences were statistically significant. The results of linear regression analysis showed that the difference in the number of TNF-α + cells in the RIRI group and the NAS group was negatively correlated with the difference in the number of Nrf2 + and HO-1 + cells ( r 2=0.923, 0.936; P<0.01). Conclusions:NAS can inhibit the expression of TNF-α protein in the retina of RIRI rats and reduce RIRI. The mechanism may be related to the Nrf2/HO-1 pathway.

Objective To explore the relationship between the polymorphisms of the primary gout predisposing gene (BCAS3) rs11653176 locus and the incidence of gout in Han Chinese men in coastal areas of Shandong Province. Methods One hundred and fifty-two cases of patients with gout remission,68 cases of acute stage,252 patients with hyperuricemia, and 280 healthy subjects, total males, were enrolled. Genotyping the rs11653176 locus of BCAS3 gene by TaqMan probe technique. The expression level of BCAS3 gene mRNA in each PBMC was measured by RT-qPCR. The levels of tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), and interleukin-18(IL-18)in serum were measured by ELISA. Results The change of allele frequency of rs11653176 locus in BCAS3 gene was associated with gout(P<0.01). BCAS3 mRNA in patients with gout was significantly higher than that of healthy people and patients with hyperuricemia(P<0.01). In gout patients, the expression level of BCAS3 gene containing C allele was higher than that of T allele(mRNA,P<0.05). The inflammatory factors in the acute phase of gout were significantly higher than those in phases of remission and hyperuricemia(P<0. 01). Conclusion Changes in the allele frequency of BCAS3 alleles rs11653176(high C, low T)may contribute to the expression of this gene,and lead to gout. And the onset of gout is closely related to the production of inflammatory factors.

Objective To evaluate the resistance of multidrug-resistant Mycobacterium tuberculosis ( M. tb) strains to bedaquiline ( BDQ) and to analyze the relationships between their genotypes and BDQ-re-sistant phenotypes in order to provide a scientific basis for rational use of BDQ for the treatment of multidrug-resistant tuberculosis ( MDR-TB) in clinical practice. Methods A total of 387 clinical M. tb strains, inclu-ding 100 pan-susceptible strains and 287 strains isolated from patients with MDR ( MDR-TB strains) , were enrolled in this study. Of the 287 MDR-TB strains, 77 strains were collected in Chongqing in 2015 and the other strains were collected in a national drug-resistant tuberculosis survey conducted in China during 2007 to 2008. Minimum inhibitory concentrations (MIC) of BDQ against those strains were detected. Genotypes of those strains were analyzed by Spoligotyping. Differences in the resistant rates against BDQ between Beijing genotype and non-Beijng genotype MDR-TB strains were comparatively analyzed. Results MIC50 and MIC90 of BDQ against the 287 MDR-TB strains were 0. 03 μg/ml and 0. 25 μg/ml, respectively. Nineteen out of the 287 MDR-TB strains (6. 6%) were resistant to BDQ. Based on the Spoligotyping, 195 strains were clas-sified into Beijing genotype, and the other 92 strains belonged to non-Beijing genotype. Statistical analysis revealed that the BDQ-resistant rate in Being genotype strains (4. 6%, 9/195) was lower than that in non-Beijing genotype strains (10. 9%, 10/92, χ2=3. 955, P=0. 047). In addition, the MIC50 and MIC90 of BDQ against pan-susceptible strains were 0. 03 μg/ml and 0. 12 μg/ml, respectively. Sixty-three pan-sus-ceptible strains belonged to Beijing genotype and the other 37 strains belonged to non-Beijing genotype. None of the pan-susceptible strains was resistant to BDQ. Conclusion This study indicates that BDQ showed stronger in vitro antibacterial activity against the MDR-TB strains isolated in China. A correlation between non-Beijing genotype and BDQ resistance is observed in those MDR strains. MDR strains of Beijing genotype are more susceptible to BDQ than those of non-Beijing genotype.

Objective To explore the construct validity of the competency assessment questionnaire for junior registered nurses and offer evidence for further tool developments. Methods A total of 603 junior registered nurses from eight level three class A hospitals in Beijing were included and investigated by the competency assessment questionnaire. Exploratory factor analysis was used to analyze the data. According to the results and the experts panel discussion, the items were selected and adjusted. Results The final questionnaire consisted of 4 factors with 23 items and the accumulating explanatory rate of the 4 factors was 61. 785%. The Cronbach′s α coefficient was 0. 787. Conclusions The competency assessment questionnaire for junior registered nurses can evaluate the competency of nurses from 4 aspects, and can also help to guide the improvement of following standardized training.

Objective To assess the changes of humidity and ammonia concentration in rat and mouse individually ventilated cages (IVC) based on macroenvironmental humidity and air ventilation changes .Methods Three kinds of rat and mouse IVC in barrier facilities were set as research objective .The changes of micronvironmental humidity and ammonia concentration at 40 times/h and 60 times /h air changes were detected continuously for a 7-days-cycle relative to low (40%), moderate (50%), and high (60%) macroenvironmental humidity.Results Mouse and rat IVC with 40 times /h air changes under low macroenvironmental humidity condition , mouse IVC with 40 times/h and rat IVC with 60 times/h air changes under moderate macroenvironmental humidity condition , mouse IVC with 60 times /h air changes under high macroenvironmental humidity condition , basically meet the GB14925-2010 requirements.While under macroenvironmental high humidity condition, the microenvironments of rat and mouse IVC with 60 times/h air changes could not satisfy the requirements.Conclusions The environmental humidity and ventilation frequency are the key index of IVC microenvironment.Only on the basis of external environment conditions to set up reasonable IVC ventilation frequency in order to better maintain the IVC microenvironment so that to achieve the goal of effective management .

Objective To determine the influence of gender on the association between serum uric acid and branchial-ankle pulse wave velocity (baPWV).Methods Cross-sectional data from 1418 adults who underwent routine laboratory tests and baPWV measurements were analyzed in a gender-specific manner.BaPWV≥ 1400 cm/s was defined as abnormality.The individuals were assigned to four groups (Q1,Q2,Q3 and Q4) according to the gender-specific quartiles of serum uric acid.The relationship between serum uric acid and the baPWV values was assessed by Spearman's correlation coefficients.In female,the relationship of serum uric acid quartiles with baPWV was evaluated by logistic regression analysis.Results In male and female,diastolic blood pressure,fasting blood glucose,triglyceride and body mass index were increased with serum uric acid level,in contrast to high-density lipoprotein cholesterol.The Spearman's correlation coefficient for the relationship between serum uric acid and baPWV was 0.18 in female and-0.05 in male (P =0.16).In female,univariate logistic regression analysis showed that in comparison with group Q1,odds ratio (OR) value of baPWV abnormality of group Q4 was 2.45 (95％confidence interval (CI) 1.48 to 4.06).In multivariate logistic regression,after adjusting for creatine and estimated glomerular filtration rate,OR value of baPWV abnormality of group Q4 was 2.45 (95％ CI 1.48 to 4.06) when compared with group Q1.However,after adjustment for age and other risk factors of cardiovascular diseases,there were no significant differents in ORs among serum uric acid quartile groups.Conclusions The association between serum uric acid level and baPWV in female may depend on age and metabolic syndrome components.

Objective To compare the efficiency of intra-articular injection of tranexamic acid (TXA) and autolo-gous transfusion drain on the blood loss after total knee arthroplasty (TKA). Methods A total of 124 patients (124 knees) with varus knee osteoarthritis, who were performed TKA,were retrospectively analyzed. Patients included 24 males and 100 females. The mean age was(65.03±6.84)years. Due to the blood loss control method, patients were divided into two groups including TXA application group (test group, n=49) and autologous transfusion drain group (control group, n=75). The data of blood routine examination, blood loss and blood transfusion after TKA were analyzed. Results The blood transfusion rates were 10.20%(5/49) in test group and 17.33%(13/75) in control group 7 days after TKA surgery. There was no signifi-cant difference between two groups (P>0.05). There were significant differences in red blood cell (RBC), hemoglobin (Hb) and haematocrit (HCT) between groups and effects of interaction in the two groups (P＜0.01). There were significantly higher values of RBC, Hb and HCT at 1, 3 and 7 days after surgery in test group than those of control group (P＜0.05).The total blood loss 1 and 3 days after TKA was significantly lower in test group than that of control group (P＜0.05). There was no sig-nificant difference in the blood loss 7 days after surgery between two groups (P>0.05). Conclusion The single dose intra-articular injection of tranexamic acid is a safe and effective procedure for hemostasis after TKA.