Abstract: Objective　To investigate the antimicrobial activity of omadacycline (OMC) against clinical Streptococcus agalactiae (GBS) isolates, as well as its relationship with biofilm formation, resistance genes and virulence genes. Methods　A total of 136 strains of Streptococcus agalactiae isolated from Shenzhen Nanshan People's Hospital between 2015 to 2020. The minimum inhibitory concentration (MIC) of OMC against Streptococcus agalactiae was determined by broth microdilution. Crystal violet staining was used to detect the biofilm formation ability of GBS. Resistance genes (tetM, tetO, tetK, ermB, OptrA) and virulence genes (cpsⅢ, bca, fbsA, cpsA, scpB) were investigated by polymerase chain reaction (PCR). Results　Among the 136 clinical isolates of GBS, 20 strains (14.7%) were resistant to OMC, 64 (47.1%) were intermediate, and 52 (38.2%) were sensitive. Fifty-seven strains (41.9%) were biofilm-positive, 20 of which (35.1%) were sensitive to OMC. Seventy-nine strains (58.1%) were biofilm-negative, 32 of which (40.5%) were susceptible to OMC. There was a statistically significant difference in the sensitivity rates between the two groups of strains (χ2=63.062, P<0.001), but there was no significant difference in the sensitivity of OMC among the biofilm-positive strains (Fisher's exact test, P=0.824). The resistance rates of tetM, tetO, ermB and OptrA positive strains were higher than those of negative strains, while tetK was opposite. The presence of tetM (Z=0.815, P=0.415), tetO (Z=0.151, P=0.88), tetK (Z=0.567, P=0.571), ermB (Z=1.198, P=0.231) resistance genes in Streptococcus agalactiae had no significant impact on the sensitivity of OMC. However, the presence of the OptrA resistance gene showed a statistically significant effect on the sensitivity of OMC (Z=2.913, P=0.004). The virulence factors cpsⅢ, bca, fbsA, cpsA and scpB were all detected at a rate higher than 50%. The presence of the virulence genes cpsⅢ (Z=0.222, P=0.824), bca (Z=0.141, P=0.888), fbsA (Z=0.813, P=0.416), and cpsA (Z=1.615, P=0.106) in Streptococcus agalactiae had no significant impact on the sensitivity of OMC. However, there was a significant inter-group difference in the scpB virulence gene (Z=2.844, P=0.004), but the rank mean values and resistance rates of scpB-positive strains were lower than those of the negative strains. Conclusions　The formation of biofilm in Streptococcus agalactiae reduces its sensitivity to OMC, but there was no significant difference in the sensitivity to OMC among the biofilm-positive strains. The presence of resistance genes tetM, tetO, tetK, ermB, and virulence genes cpsⅢ, bca, fbsA, cpsA, scpB in Streptococcus agalactiae is not associated with OMC resistance, but the presence of the resistance gene OptrA is correlated with OMC resistance..

Objective:To investigate the correlation between time within the glucose target range(TIR) and hyperuricemia(HUA) in patients with type 2 diabetes mellitus(T2DM).Methods:A total of 215 patients with T2DM in the First Affiliated Hospital of Bengbu Medical College from June 2021 to May 2022 were selected and divided into HUA group and non-HUA group according to serum uric acid level. The clinical characteristics and biochemical indicators of the patients were collected. The association of 72 h glucose monitoring system(FGMS) related indicators TIR, mean blood glucose fluctuation range(MAGE), blood glucose variability(CV), blood glucose standard deviation(SDBG), and mean blood glucose(MBG) with serum uric acid level was analyzed. The influencing factors of T2DM combined with HUA were analyzed with binary logistic regression, and receiver operating characteristic(ROC) curve was drawn to evaluate their predictive values.Results:TIR of HUA group was significantly decreased compared with non-HUA group, while HbA 1C, MAGE, CV, SDBG, and MBG were increased( P<0.001). Spearman correlation analysis showed that serum uric acid levels were negatively correlated with TIR, but positively correlated with MAGE, CV, SDBG, and MBG( P<0.001). After dichotomous logistic regression analysis, TIR was found to be an independent protective factor for T2DM with HUA. The ROC curve results showed that the area under the curve(AUC) of TIR for predicting HUA in T2DM was 0.856(95% CI 0.803-0.909, P<0.001), with the best cut-off value being 64.5%, the sensitivity being 76.8%, and the specificity being 90.3%. Conclusion:TIR in patients with T2DM combined with HUA was significantly decreased. TIR is an independent protective factor for T2DM combined with HUA, and TIR shows a certain predictive value for T2DM combined with HUA.

Objective:To summarize the survival rate, complications, and outcomes of 32 periviable extremely preterm infants (PEPIs) born at ≤23 gestational weeks.Methods:This was a retrospective observational study involving PEPIs born at the Shenzhen Maternity & Child Healthcare Hospital from January 1, 2015, to December 31, 2021. Clinical data of all subjects were collected and analyzed. The survival rates of PEPIs born from 2015 to 2019 and 2020 to 2021 were compared. Chi-square (or Fisher's exact) test was used for statistical analysis. Results:(1) During the study period, 32 PEPIs were admitted, accounting for 0.024% (32/132 534) of all newborns born in the same hospital during the study period. The median gestational age of the 32 PEPIs was 23 weeks (21 +4-23 +6 weeks), and the birth weight was 480 g (350-720 g). (2) The survival rate of PEPIs born between 2020 and 2021 was 10/19, which appears to be a trend higher than that between 2015 and 2019 (3/13, χ2=2.79, P=0.095), while the rate of withdrawal of treatment was 8/13 and 3/19, respectively, with a statistically significant difference ( χ2=7.16, P=0.007). (3) Thirteen of the 32 PEPIs survived on discharge, including four born at 22 weeks and nine at 23 weeks. The birth weights of these surviving infants were 300-<400 g in one case, 400-<500 g in five cases, 500-<600 g in four cases, 600-<700 g in one case, and ≥700 g in two cases. (4) The most common complication was moderate and severe bronchopulmonary dysplasia (10/13), followed by retinopathy of prematurity requiring surgical intervention (5/13), patent ductus arteriosus requiring ligation (4/13), late-onset sepsis (2/13), necrotizing enterocolitis (stage Ⅱa or above) (2/13) and grade Ⅲ-Ⅳ intraventricular hemorrhage or periventricular leukomalacia (2/13). The median duration of follow-up was ten months (6-69 months), and motor retardation occurred in three infants. Conclusions:The overall survival rate of PEPIs in our hospital is relatively high, with a lower incidence of complications during hospitalization and relatively better outcome. However, further studies are required for the long-term prognosis in this group of infants.

Objective:To investigate the related mechanism of IL-17B in regulating host immune response by studying the role and mechanism of IL-17B in the infection of Listeria monocytogenes in mice. Methods:Eighteen male C57BL/6 mice were randomly divided into three groups with six in each group: control group, PBS group and wild-type (WT) group. The control group was not given any treatment. The mice in the PBS group were injected with 100 μl of sterile PBS, while C57BL/6 mice in the WT group and IL-17B deficient (IL-17B -/-) male mice were injected intravenously with 100 μl of Listeria monocytogenes 19115 (2×10 4 colony forming unit). The mice were sacrificed 48 h after infection and then peripheral blood, spleen and liver samples were collected. Bacterial colonization in mouse spleen and liver was detected by plate count method; HE staining was used to evaluate histopathological damages; flow cytometry was used to detect the immune cells in different tissues. ELISA and qRT-PCR were used to detect the levels of IL-1β, IL-6, IL-12p40, TNF-α, IFN-γ and iNOS in serum and spleen. qRT-PCR were used to detect the expression of IL-17B and IL-17RB. Results:Bacterial colonization in mouse spleen was reduced in the IL-17B -/- group as compared with that in the WT group ( P<0.05). Compared with the PBS group, Listeria monocytogenes infection increased the expression of IL-17B and IL-17RB in mouse spleen ( P<0.05, P<0.01). There was no significant difference in the pathological damages in spleen between WT and IL-17B -/- groups. Moreover, compared with the WT group, the IL-17B -/- group showed increased macrophages, M1 macrophages ( P<0.01) and NK cells ( P<0.05) in spleen, up-regulated macrophages ( P<0.05) and M1 macrophages ( P<0.01) in the peripheral blood, enhanced expression of IL-6 in serum and spleen ( P<0.05), and promoted expression of IL-6, IL-12, IL-1β, TNF-α, IFN-γ and iNOS in spleen. Conclusions:IL-17B might inhibit Listeria monocytogenes clearance by inhibiting macrophage infiltration and the secretion of IL-6.

Objective:To investigate the effects of IL-28B in a mouse model of dextran sulfate sodium (DSS)-induced colitis and to analyze the possible mechanism.Methods:Thirty-five male C57BL/6 mice were randomly divided into the following groups with seven mice in each group: control group, DSS group and three IL-28B groups (1.25 μg, 2.5 μg and 5 μg). The mice in the DSS group and IL-28B groups were fed with 2.5% DSS solution and from day 3, the IL-28B groups were given intraperitoneal injection of corresponding IL-28B every day and the DSS group was treated with PBS. During the experiment, the disease activity index (DAI) was evaluated daily. On day 8, the mice were sacrificed and peripheral blood, spleen, mesenteric lymph node and colon samples were collected. The colon samples were observed, measured in length and stained with HE, and histopathological scores were calculated based on HE staining. Changes of immune cells in different samples were detected by flow cytometry. ELISA was used to detect the expression of IL-12, IL-10, IL-1β, IL-6, IL-4 and IL-13 in serum and colon tissues.Results:Compared with the DSS group, the IL-28B group (2.5 μg) had lower DAI scores [(9.40±1.67) vs (3.50±1.73), P<0.01], less shortening of the colon [(5.16±0.61) cm vs (6.91±0.60) cm, P<0.01] and significantly lower histopathological scores [(7.33±0.58) vs (4.33±0.58), P<0.01]. Moreover, compared with the DSS group, the IL-28B group (2.5 μg) showed decreased macrophages in the peripheral blood [(21.39±3.21)% vs (15.63±2.98)%, P<0.05] and spleen [(3.03±0.28)% vs (2.05±0.48)%, P<0.05], and significantly increased mean fluorescence intensity of M2 macrophages in the colon [(1 361.00±293.40) vs (2 074.00±87.61), P<0.05]. IL-12 expression in colon tissues and IL-1β expression in serum were reduced, and IL-10, IL-4 and IL-13 expression in colon tissues was significantly increased in the IL-28B group (2.5 μg) as compared with those in the DSS group [IL-12: (31.72±6.92) pg/mg vs (5.41±3.41) pg/mg; IL-1β: (48.01±16.13) pg/ml vs (12.27±6.26) pg/ml; IL-10: (184.70±46.82) pg/mg vs (444.30±157.80) pg/mg; IL-4: (2.23±0.27) pg/mg vs (3.64±0.80) pg/mg; IL-13: (11.79±0.99) pg/mg vs (22.59±1.92) pg/mg; all P<0.05]. Conclusions:IL-28B might alleviate the severity of acute enteritis in mice by increasing the secretion of IL-4 and IL-13, regulating macrophage differentiation and modulating the expression of inflammatory factors.

This study was conducted between November to December 2020, consisting of six representative cities, Beijing, Shanghai, Shenzhen (with comprehensive smoke-free legislation), and Changsha, Chongqing, Shenyang (without comprehensive smoke-free legislation), 678 subjects were enrolled eventually, the mean age of the 678 subjects was (35.61±12.91)years old. Subjects from cities with comprehensive smoke-free legislation accounted for 49.71% of the total; male subjects accounted for 19.47%; meanwhile subjects from large, medium, and small restaurants accounted for 13.57% (92), 37.32% (253) and 49.11% (333) respectively. The analysis results indicate that the positive rate of restaurants staff of cotinine and 3′-hydroxynicotinine was lower in cities with comprehensive smoke-free legislation(34.12% vs 68.04%, χ2=78.01, P<0.001; 16.32% vs 41.94%, χ2=53.79, P<0.001), with staff from cities with comprehensive smoke-free legislation have lower concentrations of cotinine and 3′-hydroxynicotinine than their counterparts from cities without comprehensive smoke-free legislation(0.250 ng/ml vs 0.742 ng/ml, P<0.001; 0.250 ng/ml vs 0.250 ng/ml, P<0.001). No statistically significant difference in the concentration of cotinine and 3′-hydroxynicotinine in saliva between staff from restaurants of different sizes was detected ( P>0.05).

This study was conducted between November to December 2020, consisting of six representative cities, Beijing, Shanghai, Shenzhen (with comprehensive smoke-free legislation), and Changsha, Chongqing, Shenyang (without comprehensive smoke-free legislation), 678 subjects were enrolled eventually, the mean age of the 678 subjects was (35.61±12.91)years old. Subjects from cities with comprehensive smoke-free legislation accounted for 49.71% of the total; male subjects accounted for 19.47%; meanwhile subjects from large, medium, and small restaurants accounted for 13.57% (92), 37.32% (253) and 49.11% (333) respectively. The analysis results indicate that the positive rate of restaurants staff of cotinine and 3′-hydroxynicotinine was lower in cities with comprehensive smoke-free legislation(34.12% vs 68.04%, χ2=78.01, P<0.001; 16.32% vs 41.94%, χ2=53.79, P<0.001), with staff from cities with comprehensive smoke-free legislation have lower concentrations of cotinine and 3′-hydroxynicotinine than their counterparts from cities without comprehensive smoke-free legislation(0.250 ng/ml vs 0.742 ng/ml, P<0.001; 0.250 ng/ml vs 0.250 ng/ml, P<0.001). No statistically significant difference in the concentration of cotinine and 3′-hydroxynicotinine in saliva between staff from restaurants of different sizes was detected ( P>0.05).

Objective To study the effect of the reinforcing and reducing method of acupuncture reported in first Chapter the Nine Needles and Twelve Yuan of the Miraculous Pivot on interstitial fluid pressure (IFP) in subcutaneous tissue of minipig, and to investigate its biomechanical mechanism of regulating the interstitial fluid. Methods Nine healthy minipigs were randomly selected for reinforcing method (pull or press) and reducing method (wave a big pinhole), and tested on soft skin tissues of the abdomen. The IFP in the normal state (NS), the low volume (LV) state (by extracting interstitial fluid) and the high volume (HV) state (by injecting saline solution) was measured before and after acupuncture. Results In the normal state, pulling and pressing the needle could obviously increase IFP, while reducing method could significantly decrease IFP, leading to a rapid decrease in 5 min after acupuncture. In the LV state, pulling and pressing the needle could increase IFP. However, in 10 min after acupuncture, the descend rates of IFP were relatively slower. In the HV state, the reducing method could significantly decrease IFP, and the changing trend in 5 min after acupuncture was different from that of the control group. Conclusions The reinforcing and reducing method of acupuncture could increase or decrease IFP, which proved that the acupuncture method could regulate IFP in the opposite direction. The research findings provide a new scientific basis for using reinforcing and reducing method of acupuncture in clinic.

Objective:To study clinical pathological characteristics, immunohistochemical, molecular genetical changes and prognosis in pediatric eosinophilic solid and cystic renal cell carcinoma (ESC RCC) with TSC2 gene mutations.Methods:The tissue samples were collected from two pediatric ESC RCC patients between 2017 and 2018. The tissues were subjected to histological examination and immunohistochemistry using EnVision system. The TFE3, TFEB gene rearrangements were tested using FISH and molecular genetic study. The paraffin sections were used for DNA extraction, PCR amplification and NGS sequencing.Results:The two patients with ESC RCC were both male, aged at 9 years and 8 months, and 13 years, respectively. The tumors were from the right kidney, 5 cm and 7 cm in size, respectively, with solid and cystic changes in cross section, and grey-reddish or grey-whitish fish meat appearance. Microscopic observation revealed the tumors had fibrous capsules, which were infiltrated by the tumor cells. The tumor cells were diffusely distributed, round-shaped, or polygon-shaped, and had voluminous cytoplasm, eosinophilic cytoplasm, various sizes of vacuoles and clear cell-like appearance. There were papillary structures in some areas, with visible fiber septa. The nuclei were round and vesicular, with multi-nucleated cells and megakaryocytes. The mitoses were not seen. A few cystic structures were visible in different sizes, and capsule walls were covered with a single layer of spike-like tumor cells. Thick-walled blood vessels were seen in the stroma, with focal lymphocytic infiltration, eosinophilic necrosis, calcifications and cholesterol crystals. Immunohistochemistry of the tumor cells was positive for PAX8 (diffuse), CK20 (focal), CKpan (focal), CK10 (1 focal, 1 diffuse), INI1, vimentin, CD68, and Ki-67 (5%~10%); the tumor cells were negative for HMB45, S-100, Melan A, p53, desmin, TFE3, CK7, CK19, EMA, CD56, CgA, Syn, CD30, CD117, WT1 and SMA. Molecular genetic study showed that TFE3 and TFEB gene rearrangements were not detected by FISH. NGS sequencing showed TSC2 p.Lys574Ter (0.198) was found in patient one and TSC2 p.Arg406Ter (0.355) in patient two.Conclusions:ESC RCC in children is a rare disease, and can be misdiagnosed easily. It has unique pathological characteristics, and immunohistochemical, molecular and genetic changes. The prognosis is relatively good.

Alzheimer' s disease is the most common kind of dementia without effective treatment. Via early diagnosis, early intervention after diagnosis is the most effective way to handle this disease. However, the early diagnosis method remains to be studied. Neuroimaging data can provide a convenient measurement for the brain function and structure. Brain structure network is a good reflection of the fiber structural connectivity patterns between different brain cortical regions, which is the basis of brain's normal psychology function. In the paper, a brain structure network based on pattern recognition analysis was provided to realize an automatic diagnosis research of Alzheimer's disease and gray matter based on structure information. With the feature selection in pattern recognition, this method can provide the abnormal regions of brain structural network. The research in this paper analyzed the patterns of abnormal structural network in Alzheimer's disease from the aspects of connectivity and node, which was expected to provide updated information for the research about the pathological mechanism of Alzheimer's disease.