A 36-year-old female presented with multiple dark brown papules on the chest and abdomen for more than 10 years,which had gradually increased in number.Physical examination revealed dozens of dark brown,flat papules measuring 1-3 mm in diameter in the chest and abdomen.Most of the lesions had a smooth surface,and some lesions gave a papilloma-like appearance,with no confluent trend.Biopsy of abdominal lesions showed mild hyperkeratosis of epidermis,acanthosis,extension of epidermal protrusions forming a reticulated appearance,horn pseudocysts in prickle cell layer,enhanced pigmentation of basal layer,and a sparse lymphocytic perivascular infiltrate in superficial dermis.A diagnosis of dermatosis papulosa nigra (DPN) was made.

Objective To study the ability of standard strain and clinical isolates of Ureaplasma spp. to form biofilms in vitro and to compare the antibiotic susceptibility of sessile cells and their planktonic counterparts. Methods A total of 21 Ureaplasma wealyticum(Uu) isolates recovered from female patients diagnosed with cervicitis and Uu serovar 3 and Uu serovar 8( Uu3, Uu8) were included. Scanning electron microscope and confocal scanning laser microscopy were used to identify biofilm formation. Conventional antibiotic susceptibility tests and biofilm susceptibility assays for tetracycline, erythromycin and ciprofloxacin were carried out. The paired rank sum test and was applied to analyze the statistical differences between the MIC and the minimal biofilm inhibitory concentration. The x2 test was applied to analyze the statistical differences of global resistance percentages between planktonic cells and sessile cells. Results Uu3, Uu8 and 21 Uu isolates all can form biofilms in vitro. Minimal inhibitory concentration of sessile cells compared with planktonic cells were obviously higher for tetracycline, erythromycin and ciprofloxacin (P <0.001). Global resistance percentages between planktonic cells and sessile cells were different for erythromycin (9.52% vs 61.90% , P < 0. 001), ciprofloxacin ( 80. 95% vs 100% , P = 0. 035 ) and tetracycline (4. 76% vs 14.29% , P =0.293). Conclusion Uu isolates and Uu1, Uu8 all can form biofilms in vitro, and biofilm formation can strengthen resistance of Uu to antibiotics, even multidrug resistance was observed.

Objective To evaluate the effects of penehyclidine hydroehloride as an atropine alternative on angioearpy and glandular secretions when premedieated in ketamine complex total intravenous anesthesia(TIVA)in children.Methods Forty patients aged 3-10 years undergoing ketamine and propofol complex TIVA were randomly divided into two groups.Penehyclidine hydrochloride(group P,n=20)or atropine(group A,n=20)was premedicated intramuscularly 30 min before anesthesia.Heart rate(HR),mean arterial pressure(MAP),breath rate(R)and the amount of saliva secretion(SS)were recorded before premedication(0 min),10 min,20 min,30 min,60 min and 150 min after.Results (1)SS reduced significantly 20 min,30 min and 60 min after premedication in both groups(P＜0.01),and in 150 min,it was still in a significantly reduced level in group P(P＜0.01),which was significantly lower than that in group A(P＜0.01).(2)MAP,HR and R in group P showed no significant differences before and after premedication(P＞0.05).But in group A,HR increased significantly at 20 min,30 min and 60 min after premedication(P＜0.05 or＜0.01),MAP increased significantly at 30 min and 60 min after premedication(P＜0.01),and meanwhile of them were also significantly higher than those in group P(P＜0.05 or＜0.01).Conclusions Penehychdine hydrochloride can effectively reduce respiratory glandular secretion with longer persistence,and nearly has no influence on HR and blood pressure,which suggests it could be a superior to atropine alternative as an anesthesia premedication in children.

Cyclophilin A (CypA) is found to be highly expressed in different kinds of tumor cells,which could regulate the occurrence and development of many kinds of tumor through multiple signal transduction pathways such as inducing the formation of inflammatory carcinoma,accelerating the transcription cycle of tumor cells,promoting the invasion and metastasis of tumor cells,inhibiting the apoptosis of tumor cells and reducing the sensitivity of tumor cells to chemotherapy drugs.It suggests that CypA might be considered as a kind of oncogene,which is expected to be a novel target for tumor treatment.

For effective inhalable dry-powder drug delivery, tetrandrine-PLGA (polylactic-co-glycolic acid) nanocomposite particles have been developed to overcome the disadvantages of nanoparticles and microparticles. The primary nanoparticles were prepared by using premix membrane emulsification method. To prepare second particles, they were spray dried. The final particles were characterized by scanning electron microscopy (SEM), dry laser particle size analysis, high performance liquid chromatography (HPLC), X-ray diffraction (XRD), differential scanning calorimetry (DSC), infrared analysis (IR) and confocal laser scanning microscope (CLSM). The average size of the primary particles was (337.5 ± 6.2) nm, while that second particles was (3.675 ± 0.16) μm which can be decomposed into primary nanoparticles in water. And the second particles were solid sphere-like with the drug dispersed as armorphous form in them. It is a reference for components delivery to lung in a new form.

Objective To investigate the extracellular polysaccharide distribution and components of Ureaplasma urealyticum (Uu) after biofilm having been developed in.Methods The standard serotype 3 and serotype 14 belong to biovar Parvo,and the standard serotype 4 and serotype 8 belong to biovar T960 were employed to form biofilrns in vitro.Scanning electron microscope and confocal laser scanning microscope were used to analysis the biofilms and extracellular polysaccharide.We used combination of two different labeled lectins,Canavalia ensiformis(FITC-ConA) and Erythrina cristagalli(ECA) which bind to specific polysaccharide residues to visualize extracellular polysaccharide in biofilms,and average uorescence intensity was evaluated Results All the strains can form the biofilmsin vitro.The biofilm was honeycomb-Like structures mainly,and extracellular polymeric substances accounts for majority of proportions.All the extracellular polysaccharide could be combined with FITC-ConA and ECA,and the total average fluorescence intensity of FITC-ConA was higher than ECA( P＜0.001 ).Conclusion Ureaplasma urealyticum biofilm is honeycomb-like structures mainly.The extracellular polysaccharide contains,galactose,and N-acetyl glucan residual,and the glucose,mannose residual are the main components.

Objective:To explore the relationship between urinary exosomal microRNA (exo-miR) - 155 in patients with type 2 diabetes mellitus (T2DM) and the onset and severity of diabetic kidney disease (DKD).Methods:From January to May 2019, 5 patients with T2DM normoalbuminuria and 5 patients with type 2 DKD were recruited from the Department of Endocrinology and Metabolism of Chongming Hospital of Shanghai Health Medical College as a microRNA screening cohort. Urine samples were collected to extract urinary exosomes, and the urine exo-miR spectrum was detected and analyzed using the miRCURY LNA array. From June 2019 to October 2022, 351 patients with T2DM who met the enrollment criteria and were matched by age and sex were included in the validation cohort in the Department of Endocrinology and Metabolism of the hospital. Patients were divided into 3 groups according to urinary albumin/creatinine ratio (UACR): normoalbuminuria group (UACR<30 mg/g, n=143), microalbuminuria group (30 mg/g≤UACR≤300 mg/g, n=171) and macroalbuminuria group (UACR>300 mg/g, n=37). According to DKD diagnostic guidelines, microalbuminuria group and macroalbuminuria group were classified into DKD group. Real-time fluorescence quantitative PCR was used to detect the expression level of exo-miR-155 in urine. Results:The results of transmission electron microscopy, nanoparticle tracking analysis, and Western blotting showed that the extraction of exosome vesicles was successful. In the screening cohort, according to the screening criteria of P<0.05 and fold changes (FC)>1.5, 226 differentially expressed miRNAs were identified from the urinary exosomes of the DKD group compared to the T2DM group. Among them, miR-155 ranged highest (FC=32.75, P<0.001). In the validation cohort, compared with the normoalbuminuria group [0.76 (0.55, 0.95)], the macroalbuminuria group [1.84 (1.18, 2.42)] had the most significant increase in urinary exo-miR-155 level ( Z=-7.411, P<0.001), followed by the microalbuminuria group [0.86 (0.69, 1.25)] ( Z=-4.092, P<0.001), and the urinary exo-miR-155 level in the macroalbuminuria group was significantly higher than that in the microalbuminuria group ( Z=-5.841, P<0.001). The correlation analysis showed that urinary exo-miR-155 level was positively correlated with UACR ( r s=0.329, P<0.001) and negatively correlated with estimated glomerular filtration rate ( r s=-0.249, P=0.015). The results of receiver operating characteristic curve analysis showed that the area under the curve of urinary exo-miR-155 level predicted DKD progression in T2DM patients was 0.892 (95% CI 0.859-0.925), corresponding cutoff value was 0.982, and the sensitivity and specificity were 71.9% and 87.7%, respectively. Multivariate logistic regression analysis showed that urinary exo-miR-155≥0.982 was an independent risk factor for progression to DKD in T2DM patients ( OR=3.310, 95% CI 1.981-5.530, P<0.001). Conclusion:The expression level of urinary exo-miR-155 is increased in T2DM patients with microalbuminuria and macroalbuminuria, which is related to the degree of albuminuria, and can be used as a predictive marker to identify potential DKD.

OBJECTIVE：To study the improvement effect and mechanism of MEBO on lipopolysaccharide （LPS）-induced injury of rat skin fibroblasts. METHODS ：Skin fibroblasts of rats were divided into control group ，LPS group （5 μg/mL）， Kangfuxin solution group （positive control ，5 μg/mL LPS+1.25％ Kangfuxin solution ）and MEBO group （5 μg/mL LPS+0.6 mg/mL MEBO），with 6 wells in each group. Inflammatory injury cell model was induced by LPS （except for control group ）. After a certain period of cultivation ，the cell survival rate and cell migration rate were detected in each group. The contents of TNF-α and IL-6 in cell supernatant was detected. The localization and fluorescence intensity of IL- 6 protein were detected. The protein expression of PTEN ，p-p65，TNF-α，IL-6，PI3K and Akt in the fibroblasts were also determined. RESULTS ：Compared with control group ，survival rate of the fibroblasts was increased significantly in LPS group ，while cell migration was decreased significantly；the contents of TNF-α and IL-6 in cell supernatant as well as relative protein expression of PTEN ，p-p65，TNF-α， IL-6 and PI 3K were increased significantly （P＜0.05 or P＜0.01）；IL-6 protein mainly expressed in the cytoplasm ，and the fluorescence intensity was enhanced. Compared with LPS group ，survival rate of the fibroblasts was decreased significantly in Kangfuxin solution group and MEBO group ，while migration rate was increased significantly ；the contents of TNF-α and IL-6， relative protein expression of PTEN ，p-p65，TNF-α，IL-6（except for Kangfuxin solution group ），PI3K and Akt （except for Kangfuxin solution group ） were decreased significantly （P＜0.05 or P＜0.01），while fluorescence intensity of IL- 6 protein decreased；relative protein expression of TNF-α，IL-6，PI3K and Akt in MEBO group were significantly lower than Kangfuxin solution group （P＜0.05 or P＜0.01）. CONCLUSIONS ：MEBO can inhibit the proliferation of LPS-induced skin fibroblasts ， reduce the level of inflammatory factors and the intensity of inflammatory reaction ， which may be related to the jiang- down-regulation of PTEN/NF-κB，PI3K/Akt signaling pathway.

Based on the transcriptome analysis data of a Bacillus licheniformis, a novel bidirectional promoter was identified from the strain and its transcriptional strength was analyzed. The expression level of a Bacillus clausii derived alkaline protease gene driven by the bidirectional promoter was studied by using the known strong constitutive promoter pShuttle-09 as a control. Three recombinant expression vectors and the corresponding recombinant bacteria were constructed. Under the control of the new promoter pLA and its reverse promoter pLB, the alkaline protease expression level respectively reached 164 U/mL and 111 U/mL. The results indicated that the transcription strength of pLA was significantly higher than that of pShuttle-09 and pLB, and both the pLA and pLB promoters could initiate the expression of the alkaline protease. Thus, it provides a new expression element for the heterogenous genes in Bacillus sp. and a new idea for the co-expression of two genes in one prokaryotic strain.
Bacillus subtilis ; Promoter Regions, Genetic

The online version of the original article can be found at https://doi.org/10.1631/jzus.B2000190 Erratum to: J Zhejiang Univ-Sci B (Biomed & Biotechnol) 2020 21(10):779-795 https://doi.org/10.1631/jzus.B2000190.