Objective　To provide a data foundation not only for food safety supervision and pollution source tracing, but also for the clinical treatment of drug-resistant bacteria in barreled drinking water, the drug resistance and type Ⅲ secretion system (T3SS) virulence genes carriage of Pseudomonas aeruginosa detected in barreled drinking water in Hainan Province were investigated, and the correlative relationship between strain ribosomal subtypes and virulence genes were then discussed. Methods The drug resistance of the isolated 55 strains of Pseudomonas aeruginosa was confirmed by using VITEK 2 Compact automatic microbial drug sensitivity system, and the T3SS virulence genes ExoU, ExoS, ExoT and ExoY were amplified by PCR, bacterial strain subtypes and homology were analyzed by the RiboPrinter automatic microbial gene fingerprint identification system. Results The drug sensitivity results showed that Pseudomonas aeruginosa isolated from barreled water had relatively low drug resistance, though one strain of multidrug-resistant Pseudomonas aeruginosa was discovered, resistant to imipenem, ciprofloxacin and cefepime. The distribution of T3SS virulence genes showed four genotypic combinations: ExoT+/ExoY+/ExoS+/ExoU- (45.45%, 25/55), ExoT+/ExoY+/ExoS-/ExoU+ (34.55%, 19/55), ExoT+/ExoY-/ExoS-/ExoU+(18.18%, 10/55), ExoT+/ExoY+/ExoS+/ExoU+ (1.82%, 1/55). Ribosomal typing results showed that the Pseudomonas aeruginosa strains were divided into six subtypes, the numbers of each subtype accounted for 24 (43.64%), 1 (1.82%), 25 (45.45%), 1 (1.82%), 3 (5.45%) and 1 (1.82%) respectively, with subtype Ⅰ and subtype Ⅲ being dominant. The main T3SS genotypes of the top two subtype I and subtype III were ExoT+/ExoY+/ExoS-/ExoU+ (16/24, 66.67%) and ExoT+/ExoY+/ExoS+/ExoU- (22/25, 88%). Conclusions The T3SS secretion system exhibits the characteristics of multiple virulence genes' coordinated expression, and there is a certain correlation between subtypes of bacterial strains and virulence genotypesThe exploration of the relationship between them provides guidance for tracing the source of Pseudomonas aeruginosa contamination, production control, clinical treatment in barrelled drinking water, and preliminarily establishes the initial data of local Pseudomonas aeruginosa strains in barrelled drinking water as well the related drug sensitivity data in Hainan.

Objective:To analyze the differences in whole blood selenium (Se), zinc (Zn), copper (Cu), magnesium (Mg), calcium (Ca), and iron (Fe) levels of rural older adults among areas with different soil selenium levels, and explore the main factors associated with the six nutrient elements status, so as to provide a basis for further evaluating the health risks of people in areas with different soil selenium levels.Methods:Four administrative villages were randomly selected from the Se-deficient (soil Se content < 0.175 mg/kg), Se-sufficient (soil Se content 0.175 - < 0.400 mg/kg), Se-rich (soil Se content 0.400 - < 3.000 mg/kg) and Se-excessive (soil Se content ≥3.000 mg/kg) areas, respectively, in Enshi Tujia and Miao Autonomous Prefecture (Enshi Prefecture) of Hubei Province in 2017 - 2018. And 100 elderly people aged 60 years or older (half male and half female) were randomly selected as the survey subjects in each servey site. The basic information such as general demography and lifestyle was collected through face-to-face questionnaires. Physical examination was performed and fasting venous blood was collected in the morning. The contents of blood Se, Zn, Cu, Mg, Ca, and Fe were determined by inductively coupled plasma mass spectrometry. The main factors associated with the six nutrient elements status were analyzed.Results:A total of 416 subjects were included, including 208 males and 208 females, whose average age was (72.43 ± 5.25) years, and body mass index (BMI) was (22.67 ± 3.49) kg/m 2. There were significant differences of blood Se, Zn, Cu, Mg, Ca and Fe levels between the areas with different Se levels ( Z/F = 288.30, 3.24, 14.81, 29.14, 131.28, 3.37, P < 0.05). Compared with Se-deficient and Se-sufficient areas, blood Se level was higher in Se-rich and Se-excessive areas and blood Zn level was lower in Se-excessive area ( P < 0.05); compared with Se-sufficient area, blood Cu level was lower in Se-deficient, Se-rich and Se-excessive areas, but blood Mg and Ca levels were higher ( P < 0.05), and the blood Fe level was lower in Se-excessive area ( P < 0.05). There were significant differences in the deficiency rates of Se, Zn, Cu, Mg, Ca and Fe among the elderly in different Se level areas (χ 2 = 140.83, 15.39, 31.90, 17.49, 157.60, 30.33, P < 0.01). There were significant differences in blood Zn, Cu, Ca and Fe levels between two gender groups ( P < 0.05); the blood Zn and Fe levels of the smokers were higher than those of the non-smokers, and the blood Cu level was lower than that of the non-smokers ( P < 0.05); the blood Zn and Fe levels of the drinkers were higher than those of the non-drinkers ( P < 0.05). Conclusions:The levels of six nutrient elements in the whole blood of the elderly in areas with different soil Se levels are different. To assess the health risks of the population in areas with different soil Se levels, it is necessary to consider the levels of multiple nutrient elements at the same time.

Objective:To investigate the changes in liver function and peripheral regulatory lymphocytes before and after treatment in patients with occupational medicamentosa-like dermatitis due to trichloroethylene (OMDT) .Methods:In December 2019, 16 patients with OMDT (8 patients with erythema multiforme and 8 with erythema multiforme major) who were admitted from February 2017 to February 2019 were enrolled as subjects. Liver function parameters and percentages of peripheral regulatory lymphocytes were measured before and after treatment, and the changes in liver function and peripheral regulatory T and B lymphocytes and their correlation were analyzed.Results:Before treatment, compared with the healthy control group, the experimental group had significantly higher levels of alanine aminotransferase (ALT) , aspartate aminotransferase (AST) , total bilirubin (TBIL) , direct bilirubin (DBIL) and gamma-glutamyl transpeptidase (GGT) and significantly lower levels of total protein (TP) , albumin (ALB) and cholinesterase (CHE) ( P<0.05) . Compared with the healthy control group, the experimental group had significantly lower percentages of lymphocytes, CD4 + T cells, CD4 +CD25 + Tregs, CD19 +CD24 +CD27 + Bregs and CD4 +/CD8 + ratio, as well as a significantly higher percentage of CD8 + T cells ( P<0.05) . Before treatment, the levels of ALT, AST, GGT and DBIL were negatively correlated with the percentages of CD4 +CD25 + Tregs, CD19 +CD24 +CD27 + Bregs, CD4 + T cells and CD4 +/CD8 + ratio ( r=-0.386 to -0.809, P<0.05) and was positively correlated with the percentage of CD8 + T cells (except DBIL) ( r=0.503-0.568, P<0.05) . The levels of TP and ALB were positively correlated with the percentages of CD4 +CD25 + Tregs, CD19 +CD24 +CD27 +Bregs and CD4 + T cells ( r= 0.351-0.784, P<0.05) , ALB was negatively correlated with the percentage of CD8 + T cells ( r=-0.315, P<0.05) . CHE was positively correlated with the percentages of CD4 +CD25 + Tregs, CD19 +CD24 +CD27 +Bregs and CD4 +/CD8 + ratio ( r=0.390-0.527, P<0.05) . Conclusion:Immune dysfunction is observed in patients with OMDT, which may be caused by the imbalance of regulatory lymphocytes. And liver injury may be associated with the increase of CD8 + T cells and the reductions of percentages of CD4 + T cells, CD4 +CD25 + Tregs, CD19 +CD24 +CD27 +Bregs and CD4 +/CD8 + ratio.

Objective:To investigate the changes in liver function and peripheral regulatory lymphocytes before and after treatment in patients with occupational medicamentosa-like dermatitis due to trichloroethylene (OMDT) .Methods:In December 2019, 16 patients with OMDT (8 patients with erythema multiforme and 8 with erythema multiforme major) who were admitted from February 2017 to February 2019 were enrolled as subjects. Liver function parameters and percentages of peripheral regulatory lymphocytes were measured before and after treatment, and the changes in liver function and peripheral regulatory T and B lymphocytes and their correlation were analyzed.Results:Before treatment, compared with the healthy control group, the experimental group had significantly higher levels of alanine aminotransferase (ALT) , aspartate aminotransferase (AST) , total bilirubin (TBIL) , direct bilirubin (DBIL) and gamma-glutamyl transpeptidase (GGT) and significantly lower levels of total protein (TP) , albumin (ALB) and cholinesterase (CHE) ( P<0.05) . Compared with the healthy control group, the experimental group had significantly lower percentages of lymphocytes, CD4 + T cells, CD4 +CD25 + Tregs, CD19 +CD24 +CD27 + Bregs and CD4 +/CD8 + ratio, as well as a significantly higher percentage of CD8 + T cells ( P<0.05) . Before treatment, the levels of ALT, AST, GGT and DBIL were negatively correlated with the percentages of CD4 +CD25 + Tregs, CD19 +CD24 +CD27 + Bregs, CD4 + T cells and CD4 +/CD8 + ratio ( r=-0.386 to -0.809, P<0.05) and was positively correlated with the percentage of CD8 + T cells (except DBIL) ( r=0.503-0.568, P<0.05) . The levels of TP and ALB were positively correlated with the percentages of CD4 +CD25 + Tregs, CD19 +CD24 +CD27 +Bregs and CD4 + T cells ( r= 0.351-0.784, P<0.05) , ALB was negatively correlated with the percentage of CD8 + T cells ( r=-0.315, P<0.05) . CHE was positively correlated with the percentages of CD4 +CD25 + Tregs, CD19 +CD24 +CD27 +Bregs and CD4 +/CD8 + ratio ( r=0.390-0.527, P<0.05) . Conclusion:Immune dysfunction is observed in patients with OMDT, which may be caused by the imbalance of regulatory lymphocytes. And liver injury may be associated with the increase of CD8 + T cells and the reductions of percentages of CD4 + T cells, CD4 +CD25 + Tregs, CD19 +CD24 +CD27 +Bregs and CD4 +/CD8 + ratio.

Objective To determine the expression of miRNA-148a-3p in CD4+ T lymphocytes in peripheral blood of patients with psoriasis vulgaris,and to explore its role in occurrence of psoriasis vulgaris.Methods Totally,20 patients with psoriasis vulgaris and 20 healthy controls were enrolled from Guangzhou Institute of Dermatology between July 2017 and April 2018.Peripheral venous blood samples were obtained from these subjects,and CD4+ T lymphocytes were isolated from these peripheral blood samples by magnetic cell sorting system.Real-time quantitative PCR (RT-PCR) was performed to determine the expression of miRNA-148a-3p in CD4+ T lymphocytes in the peripheral blood.Potential target genes of miRNA-148a were predicted by using bioinformatics software,and verified by using a dual-luciferase reporter system.Western blot analysis was conducted to determine the protein expression of Bcl-2 interacting mediator of cell death (Bim,the potential target gene of miRNA-148a-3p) in the CD4+ T lymphocytes of the subjects.Statistical analysis was carried out with SPSS 20 software by two sample-t test for comparing the means of normally distributed data,and by Pearson correlation analysis for analyzing the correlation of two variables.If the data were not normally distributed,Mann Whitney U test was used for comparing means between two groups,and Spearman correlation analysis for analyzing the correlation of two variables.Results The miRNA-148a-3p expression in the CD4+ T lymphocytesin the psoriasis vulgaris group (18 cases,5.61 ± 1.66) was significantly higher than that in the healthy control group (12 cases,1.00 ± 0.26;U =12,P ＜ 0.05),and was positively correlated with the psoriasis area severity index (PASI) score (r =0.93,P ＜ 0.001).Bim was predicted to be one of the potential target genes of miRNA-148a-3p by bioinformatics software,which was also verified by using a dual-luciferase reporter system.The protein expression of Bim in the CD4 + T lymphocytes was significantly lower in the psoriasis vulgaris group (11 cases,0.69 ± 0.07) than in the healthy control group (8 cases,0.93 ± 0.06;t =4.38,P ＜ 0.01),and the protein expression of Bim in the patients with psoriasis vulgaris was negatively correlated with PASI score (r =-0.774,P ＜ 0.01).Conclusion miRNA-148a-3p is overexpressed in CD4+ T cells in the peripheral blood of patients with psoriasis vulgaris,which may regulate the protein expression of Bim,leading to abnormal activation of CD4+ T cells,and then participate in the occurrence and development of psoriasis.

Objective To evaluate the efficacy and safety of microneedling radiofrequency in the treatment of facial acne scars.Methods A retrospective study was conducted.According to the inclusion and exclusion criteria,42 patients with acne scars receiving microneedling radiofrequency treatment (observation group) and 47 patients with acne scars treated with fractional CO2 laser (control group) in Guangzhou Institute of Dermatology were enrolled into this study from June 2016 to June 2017.Single-blind evaluation was conducted according to a four-grade scoring system and ECCA grading scale (échelle d'évaluation clinique des cicatrices d'acné) by two clinically experienced dermatologists to evaluate and compare the clinical efficacy between the 2 groups,based on the clinical photos before and after the treatment.Statistical analysis was done by using chi-square test,t test of two independent samples and paired Wilcoxon signed rank test.Results After treatment,no significant difference in the total response rate was observed between the observation group (92.9％,39 cases) and control group (89.4％,42 cases;x2 =0.042,P ＞ 0.05).Before the treatment,there was no significant difference in the ECCA score between the observation group (46.7 ± 16.3) and control group (45.7 ± 15.8,t =0.271,P ＞ 0.05).After the treatment,the ECCA score in the observation group and control group significantly decreased into 29.5 ± 15.1 and 31.3 ± 14.9 respectively (Z =-5.713,-6.086,respectively,both P ＜ 0.05).Conclusion Microneedling radiofrequency is effective for the treatment of facial acne scars,with high safety and less injuries.

Objective To investigate the potential roles of pattern recognition receptor in the pathogenesis of asthma,the concentrations of serum nucleotide -binding oligomerization domain like receptors 1 (NOD1 )and Toll like receptor 1(TLR1)and Dectin and sCD14 protein were determined.Methods Blood samples were obtained from 45 cases of severe asthma(asthma group)during acute attack and convalescent period,the concentrations of serum NOD1,TLR1,Dectin and sCD14 protein were assessed using enzyme linked immunosorbent assay(ELISA),and compared with 30 healthy children(control group).Results The concentrations of NOD1,TLR1,Dectin and sCD14 protein were significantly higher in acute attack period of the asthma group[(65.53 ±19.95)ng/mL,(10.46 ±3.35)ng/mL, (80.38 ±19.51)ng/mL,(4.51 ±1.29)ng/mL]than those in the control group[(25.57 ±9.64)ng/mL,(5.54 ± 1.49)ng/mL,(47.37 ±15.16)ng/mL,(2.21 ±0.68)ng/mL](t =11.57,8.63,7.82,9.95,all P <0.01).Moreo-ver,the concentrations of serum NOD1,Dectin and sCD14 protein were dramatically decreased in convalescent period of the asthma group[(39.67 ±11.25)ng/mL,(60.04 ±16.76)ng/mL,(3.38 ±0.97)ng/mL]compared with acute asthma attack period(t =8.19,6.32,5.42,all P <0.01),while the concentration of serum TLR1 was not different between this two period (9.51 ±3.19)ng/mL(t =1.27,P >0.05).Furthermore,level of NOD1 was positively corre-lated with sCD14(r =0.49,P <0.01).Conclusion Levels of serum NOD1,TLR1,Dectin and sCD14 protein were increased in acute severe asthma,whereas,they were decreased in convalescent period.The elevation of them indicated that pattern recognition reception has synergistic function in the pathogenesis of asthma attack.

Nerve injury-induced protein-1 (Ninjurin-1, Ninj1) was initially identified as a novel adhesion molecule in rat sciatic nerve and to be up-regulated in neurons and Schwann cells of distal nerve segments after nerve transection or crush injury. Recently, Ninj1 was found to act as a modulator of cell migration, angiogenesis, and apoptosis. Accumulating evidence indicates that innate immune response plays beneficial and deleterious roles in brain ischemia, and the trans-endothelial migration of blood-derived immune cells is key initiator of this response. In the present study, we examined the expression profile and cellular distribution of Ninj1 in rat brain after transient focal cerebral ischemia. Ninj1 expression was found to be significantly induced in cortical penumbras 1 day after 60 min of middle cerebral artery occlusion (MCAO) and to increase gradually for 8 days and then declined. In infarction cores of cortices, patterns of Ninj1 expression were similar to those observed in cortical penumbras, except induction was maintained for 10 days. At 1 day post-MCAO, Ninj1 inductions were detected mainly in neutrophils and endothelial cells in both infarction cores and penumbras, but reactive macrophages were the major cellular expressers of Ninj1 at 4 days post-MCAO. Expressional induction in reactive macrophages was maintained in infarction cores after 12 days post-MCAO but not in penumbras. These dynamic expressions of Ninj1 in different immune cells at different times suggest that this protein performs various, critical roles in the modulation of acute and delayed immune responses in the postischemic brain.
Animals ; Apoptosis ; Brain Ischemia* ; Brain* ; Cell Movement ; Endothelial Cells ; Immunity, Innate ; Infarction ; Infarction, Middle Cerebral Artery ; Macrophages ; Myeloid Cells* ; Neurons ; Neutrophils ; Rats* ; Schwann Cells ; Sciatic Nerve

Nerve injury-induced protein-1 (Ninjurin-1, Ninj1) was initially identified as a novel adhesion molecule in rat sciatic nerve and to be up-regulated in neurons and Schwann cells of distal nerve segments after nerve transection or crush injury. Recently, Ninj1 was found to act as a modulator of cell migration, angiogenesis, and apoptosis. Accumulating evidence indicates that innate immune response plays beneficial and deleterious roles in brain ischemia, and the trans-endothelial migration of blood-derived immune cells is key initiator of this response. In the present study, we examined the expression profile and cellular distribution of Ninj1 in rat brain after transient focal cerebral ischemia. Ninj1 expression was found to be significantly induced in cortical penumbras 1 day after 60 min of middle cerebral artery occlusion (MCAO) and to increase gradually for 8 days and then declined. In infarction cores of cortices, patterns of Ninj1 expression were similar to those observed in cortical penumbras, except induction was maintained for 10 days. At 1 day post-MCAO, Ninj1 inductions were detected mainly in neutrophils and endothelial cells in both infarction cores and penumbras, but reactive macrophages were the major cellular expressers of Ninj1 at 4 days post-MCAO. Expressional induction in reactive macrophages was maintained in infarction cores after 12 days post-MCAO but not in penumbras. These dynamic expressions of Ninj1 in different immune cells at different times suggest that this protein performs various, critical roles in the modulation of acute and delayed immune responses in the postischemic brain.
Animals ; Apoptosis ; Brain Ischemia* ; Brain* ; Cell Movement ; Endothelial Cells ; Immunity, Innate ; Infarction ; Infarction, Middle Cerebral Artery ; Macrophages ; Myeloid Cells* ; Neurons ; Neutrophils ; Rats* ; Schwann Cells ; Sciatic Nerve

Objective Through the detections of the heterozygote frequencies tests of fetal specific genes PLAC4 and COL6A2 mRNA alleles in plasma of pregnant women, to explore its possibility of application in the noninvasive prenatal screenings of trisomy-21. Methods A toltal of 500 cases (males and females 250 cases respectively)of Han ethnic groups with Henan Provice of China who were subject to the physical checkup clinic of the Third Affiliated Hospital,Zhengzhou University from June to December, 2013 were selected as the healthy physical checkup group, and such techniques as DNA sequencing and PCR-restriction fragment length polymorphism (RFLP) were adopted to the determinations of the heterozygote frequencies of the single nucleotide polymorphism(SNP)of the PLAC4 and COL6A2 genes in the maternal peripheral blood in the healthy physical checkup group, and the differential comparisons of the determination results of the SNP heterozygote frequencies and the corresponding heterozygote frequencies in the National Center for Biotechnology Information (NCBI) database;30 cases of healthy pregnant women who spontaneously underwent pregnancy checkups at the maternity clinic were randomly selected as the healthy pregnancy group, and real-time fluorescence quantitative reverse transcription-PCR technique was adopted for determining the expression levels of PLAC4 and COL6A2 mRNA in the peripheral blood of pregnant women of 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks;40 cases of the same phase were selected for acting as the specimens for the karyotype analyses of the amniotic fluid cells, among which 20 cases were trisomy-21, and the 20 cases of the negative control group, and reverse transcription-multiplex ligation dependent probe amplification (RT-MLPA) technique was adopted for screening the fetal trisomy-21. Results (1) The allele heterozygote frequencies of the SNP of the healthy physical checkup group:determinations of the genotypes and hybrid rates of the 10 SNP sites of the PLAC4 and COL6A2 genes indicated that those with higher heterozygote frequencies were respectively rs7717, rs559, rs1044598, rs59066201 and rs1042917, with population coverage of 98%. Among them, the allele hybrid rates of rs59066201 were never seen in the NCBI database;in the respective comparisons of the allele hybrid rates of rs8130833, rs9977003 and rs7844 with the hybrid rates of the NCBI database, the variations had statistical significance (P<0.05). (2) The expression levels of PLAC4 and COL6A2 mRNA of the different pregnancy weeks of the healthy pregnancy group: the levels of PLAC4 mRNA in the peripheral blood of women of 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks of pregnancy were respectively 7.22 ± 1.05, 8.02±1.41,9.51±1.69,11.33±2.11 and 13.31±2.58, with their expression levels rising along with the increase of the pregnancy weeks; among them, the comparison of pregnancy 8 weeks and pregnancy 10 weeks, the variations had no statistical significance (P>0.05);in the mutual comparisons among the expression levels of the various pregnancy weeks, the variations had statistical significance (P<0.05). The expression levels of COL6A2 mRNA in 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks were respectively 8.95 ± 1.28, 11.19 ± 1.36,15.00 ± 1.58,16.87 ± 1.72 and 18.96 ± 2.79, with their expression levels rising along with the increase of the pregnancy weeks, and in the mutual comparisons between the expression levels of the various pregnancy weeks, the variations all had statistical significance (P<0.05). (3) Prenatal screenings of trisomy-21 in the validation group of the trisome:a total of 5 sites of rs7717, rs559, rs1044598, rs59066201 and rs1042917 were selected from the allele heterozygote frequencies of SNP sites were selected from the subjects of the healthy physical checkup group, and 10 cases of trisomy-21 specimens and 10 cases of negative CTR specimens were accurately determined, with the sensitivity reached 80%(17/20), and the specificity reached 90%(18/20). One case of the trisomy-21 and two negative cases were both homozygotes, and among the trisomy-21 specimens of two cases, only one SNP was a heterozygote, and it was impossible to conduct screenings on these 5 cases, with the screening accuracy reaching 100%(35/35). Conclusions Fetal specific genes PLAC4 and COL6A2 mRNA are expressed in the peripheral blood of pregnant women in different gestational age;its expression level increases with the increase of gestational age. Among them, five SNP including rs7717, rs559, rs1044598, rs59066201 and rs1042917 show highest heterogeneity rate, which is different from the corresponding heterogeneity rate in NCBI database. RT-MLPA technology is a rapid, effective, noninvasive and low cost method of prenatal screening 21 trisomy.