Objective To understand the mutations of macrolide resistance gene locus (23S rRNA) of Mycoplasma pneumoniae (MP) and its correlation with clinical features .Methods A total of 354 respiratory tract samples were collected from children pa-tients with pneumonia .MP and its mutations in 23S rRNA gene locus were detected by real-time PCR .The children cases of MP positive were divided into the mutation group and non-mutation group .Then the clinical data were compared between the two groups .Results Among 354 respiratory tract samples ,166 cases(46 .9% ) were MP positive ,moreover the mutation of 23S rRNA gene locus existed in 135 MP positive samples with the positive detection rate of 81 .3% ,while no 23S rRNA gene locus mutations were detected in 31 samples .Analyzing the clinical data of the mutation group and non-mutation group found that there was no sta-tistical difference in the aspects of age and gender between the two groups .The occurrence rates of severe pneumonia and extrapul-monary complications in the mutation group were higher than those in the non-mutation group (P<0 .05) ,moreover the average hospitalization time and fever duration in the mutation group were longer than those in the non-mutation group (P<0 .05) .Conclu-sion 23S rRNA gene locus mutation has higher detection rate ,prompting that MP shows high resistant rate to macrolides ,which could provide a certain basis for treatment of M P infections .

Adult ; Altitude ; Arteries ; metabolism ; Calcitonin Gene-Related Peptide ; blood ; Humans ; beta-Endorphin ; blood

Objective To observe the expression of survivin and Aurora B in human pancreatic cancer BXPC3 cells after the treatment of sulindac and to explore the potential mechanism. Methods MTr assay was used to determine the effect of sulindac on the proliferation of the BXPC3 cells. RT-PCR was used to detect the expression of mRNA level of survivin and Aurora B, western blot was used to detect protein expression of survivin and Aurora B Thr-232. Cell cycle and apoptosis were detected by flow eytometry (FCM). Results The BXPC3 cells were inhibited by sulindac in a dose and time-dependent manner; the expression of mRNA of survivin and Aurora B were both significantly decreased from 1.5644 and 0.6554 to 0. 4372 and 0.1132 (P< 0.01), the expression of survivin protein and the phosphorylation of Aurora B Thr-232 were also decreased from 1.2735 and 0.4680 to 0.2126 and 0.2546 (P<0.01); the proportion of cells in the G0/G1 phase was increased from (56.65±1.93)% to (70.58±3.21)% (P<0.01). Conclusions Sulindac had inhibitory effects on the growth of BXPC3 cells, the possible mechanism was via decreasing the expression of survivin which depressed the activity of Aurora B, then the CPC was influenced. The most of the cells were blocked in the G0/G1 phase, and the cells' mitosis was inhibited.

Objective To investigate the distribution,epidemiologic features and antibiotic resistance of the enteric pathogens i-solated from children with diarrhea.Methods Enteric pathogens were isolated from children’s stool samples.The children with diarrhea were treated in the outpatient and inpatient of Shanghai Children’s Hospital between 2008 and 2013.Antimicrobial susceptibility testing was conducted by disk diffusion method for Salmonella and Shigella with 6 antimicrobial agents.Results A total of 545 enteric pathogens were collected.Salmonella was the dominant pathogen,accounting for 67.2%,followed by Shigella (20.7%),S.aureus (4.6%),C.jejuni (3.7%),Aeromonas (2.4%),and enteropathogenic E.coli (0.9%).The main serotypes of Salmonella were S.typhimurium and S.enteritidis.Approximately 56.3% of the patients were boys.A-bout 72.7% of the patients were infants under 2 years.The prevalence of diarrhea peaked in summer and autumn (72.9%). The susceptibility of these isolates was only tested with seven antibiotics.Shigella showed higher level of resistance to ampicil-lin and trimethoprim-sulfamethoxazole than Salmonella (P<0.05).Significantly higher percentage of S.flexneri isolates were resistant to sulbactam-ampicillin,ceftriaxone,ciprofloxacin,and chloramphenicol than S.sonnei (P<0.001).Further-more,the prevalence of multidrug resistant strains in Shigella (68.3%)was much higher than that in Salmonella (44.7%,P<0.001).Conclusions A variety of diarrhea-causing enteric pathogens are isolated from the children in Shanghai Children’s Hospital.The isolates are predominantly Salmonella and Shigella species.The epidemiological features of Salmonella and Shigella species are different in terms of gender,age,season and geographical distribution.The resistance to antibiotics is a serious problem and varies with different types of pathogens. Intensive and ongoing surveillance of enteric pathogens and their changing resistant pattern is required to control diar-rhea in children.

Objective:To choose the best vector for the expression of CS3 fimbriae. Methods: The CS3 operon was cloned into different plasmid vectors such as pUC19 and pTrc99A. The expression of CS3 was monitored by whole-cell ELISA and SDS-PAGE analysis. The assembly of CS3 fimbriae was detected with electron microscopy. Results:The expression level of CS3 fimbriae using plasmid pUC19 as carrying vector was the highest, and the insertion orientation of CS3 gene into the plasmid has a little effect on its expression level. The expression of CS3 fimbriae was confirmed by SDS-PAGE analysis and electron microscopy.Conclusions:The promotor of CS3 itself played the key role in the expression of CS3 fimbriae and the copy number of plasmid was the main factor to affect the expression level.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anti-cancer agent. Epigallocatechin-3-gallate (EGCG) is a polyphenolic constituent of green tea. In this study, inhibitory effect of combined use of EGCG and TRAIL on human melanoma A375 cells was examined and the possible mechanism investigated. The cells were divided into 4 groups: control group, EGCG group (EGCG: 10, 20 mug/mL), TRAIL group (TRAIL: 25 ng/mL) and EGCG+TRAIL group (combined group). The growth inhibition was measured in the A375 cells treated with different concentrations of TRAIL ((25, 50, 75, 100, 125, 150 ng/mL) by MTT assay. The apoptosis was assessed by flow cytometry. The expressions of DR4 and DR5 were detected by flow cytometry and western blotting. The activities of caspase-8 and caspase-3 were determined by colorimetric assay. The results showed that TRAIL could dose-dependently inhibit the growth of A375 cells and the IC(50) of TRAIL was 150 ng/mL. The apoptosis rate was 11.8% in the TRAIL group, 5%-7% in the EGCG group and 48.9%-59.1% in the combined group. Significant difference was found in the apoptosis rate between the combined group and the EGCG or TRAIL group (P<0.05 for each). The expression of DR4 instead of DR5 was significantly increased in the EGCG group. The activity of caspase-3 rather than caspase-8 was substantially enhanced in the EGCG group. These results suggest that EGCG is useful for the TRAIL-based treatment for melanoma.

Several methods for the qualitative screening of fungi to degrade lignin were introduced in this paper, with detailed protocols and discussing for each assay.

Objective To investigate the changes of IL‐1β,IL‐2 ,IL‐4 ,IL‐6 ,IL‐10 and INF‐γexpressions in bone marrow of chro‐nic myeloid leukemia(CML)patients .Methods The IL‐1β,IL‐2 ,IL‐4 ,IL‐6 ,IL‐10 and INF‐γexpression levels were detected by u‐sing flow cytometry in 30 cases of CML chronic phase(CML‐CP) ,21 cases of CML accelerated phase(CML‐AP) ,15 cases of CML blastic phase(CML‐BP) ,42 cases of CML remission after treatment and 7 cases of non‐remission .Then the detection results were compared with those in the control group .Results The expression levels of INF‐γ and IL‐2 in each CML groups were lower than those in the control group(P<0 .05) ,while the expression levels of IL‐1β,IL‐4 ,IL‐6 and IL‐10 were higher than those in the con‐trol group(P<0 .05) .With the disease condition progression ,the INF‐γand IL‐2 levels were gradually decreased ,i .e .,CML‐BPCML‐AP> CML‐CP ,the difference among groups had statistical significance (P<0 .05) .After complete remission by treatment ,the INF‐γand IL‐2 levels were risen to some extent ,but which were still lower than those in the control group(P<0 .01) ,the expression levels of IL‐1β,IL‐4 ,IL‐6 and IL‐10 were decreased to some extent ,but likewise were higher than those in CML‐CP(P< 0 .05);the levels of IL‐1β,IL‐4 ,IL‐6 ,IL‐10 ,IL‐2 and INF‐γ had no statistical difference between the non‐remission group and CML‐BP group(P>0 .05) .Conclusion The changes of serum cytokines in bone marrow microenvironment of CMLpatients have certain significance to the occurrence ,development and prognosis of CML ;the de‐tection of IL‐1β,IL‐2 ,IL‐4 ,IL‐6 ,IL‐10 and INF‐γlevels in bone marrow is hopeful to provide new ideas and theoretical basis for im‐mune therapy and prognosis judgment of CML patients .

Objective To find out iodine nutritional status of pregnant women in the third trimester and the relationship between pregnant women and their neonates thyroid function in a high iodine area.Methods From April to June 2010,210 copies of fasting morning urine and venous blood,and their neonatal umbilical cord blood samples were collected in Haixing Hospital,Cangzhou city,Hebei province.Urinary iodine was determined by arsenic-cerium catalytic spectrophotometry.The levels of free triiodothyronine(FT3),free thyroxine (FT4) and sensitive thyroid-stimulating hormone (sTSH) in serum were measured by chemiluminescence.Results Median urinary iodine of 210 pregnant women(average age 27.69 ± 4.73 years) in the third trimester was 1240.70 μg/L,84.3％(177/210) of them was in excessive iodine nutrition,and only 0.5％(1/210) of them was in adequate iodine nutrition.The prevalence rate of thyroid diseases was 19.5％ (41/210),and the spectrum of diseases were subclinical hypothyroidism(16.2％,34/210),subclinical hyperthyroidism(0.9％,2/210),hypothyroidism(2.4％,5/210) and hyperthyroidism (0,0/210).The number of newborns with sTSH 5 - ＞ 10 mU/L were 104 persons (49.5％); 10 - ＞ 20 mU/L were 44 persons(21.0％),and ≥20 mU/L were 16 persons(7.6％).Of pregnant women suffer from thyroid disease,the ratio(50.0％,24/48 ) of sTSH equal to 10.18 mU/L and ＞ 10 mU/L of their neonates was higher than that of their corresponding non-ill pregnant women(6.78 mU/L,Z =- 2.867,P ＜ 0.05; 22.2％,36/162,x2 =14.000,P ＜ 0.05).There was a positive correlation between neonates' and their mothers' sTSH levels (r =0.278,P ＜ 0.05).There was also a positive correlation between neonates' (sTSH ＞ 10 mU/L) and their mothers' abnormal sTSH levels (r =0.240,P ＜ 0.05).Conclusions Most of the pregnant women in high iodine areas are iodine excess.The level of neonates' sTSH is higher,and it is due to their mothers' abnormal sTSH and suffering from thyroid diseases to some extent.As a result,the monitoring of pregnant women's iodine nutrition and thyroid function and sTSH level of their neonates should be strengthened.

Objective To investigate the correlation of plasma interleukin ( IL)-17 level and IL-17 receptor (IL-17R) expression in the thymus of patients with myasthenia gravis (MG).Methods The blood samples of 63 patients (38 with glucocorticoid treatment, 25 with thymus removal) who admitted to Henan Provincial People′s Hospital between 2010 and 2014 were collected at three different stages: pre-treatment, 1 week post-treatment and 1 month post-treatment.The blood samples of 42 healthy controls were also collected.Enzyme linked immunosorbent assay was used to evaluate the levels of IL-17 in plasma.Twenty-five thymus tissues from MG patients and another 12 thymus tissues from patients with congenital heart disease who had surgery therapy were also collected.Reverse transcription polymerase chain reaction was used to evaluate the mRNA levels of IL-17R.The possible correlation between the expression of IL-17 and IL-17R with MG was analyzed.Results Before treatment, the levels of IL-17 in the plasma were much higher in all the MG patients ( both ocular and generalized) when compared to the healthy controls ( controls (3.2 ±0.7) pg/ml, MG patients (8.5 ±1.7) pg/ml, t =2.450, P <0.01; generalized type patients (9.7 ±1.4) pg/ml, t =2.532, P <0.01).In the patients with glucocorticoid treatment, IL-17 levels began to reduce after 1 week treatment and a statistically significant difference was found when compared to the pre-treatment samples (pre-treatment (8.3 ±1.2) pg/ml, 1 week after treatment (6.3 ±0.7) pg/ml, t=2.052, P<0.05) and healthy controls (t =1.933, P<0.05).One month after the glucocorticoid treatment, the levels of IL-17 decreased to the normal level (1 month after treatment (3.9 ±0.6) pg/ml, t=2.630, P <0.01, compared to the pre-treatment; t =1.395, P >0.05, compared to the healthy controls).In the surgery therapy cases, the IL-17 levels were also reduced after the thymus removal ( pre-surgery (8.8 ±1.4) pg/ml, 1 week after surgery (5.3 ±0.7) pg/ml, t=1.950, P<0.05;1 month after surgery (3.0 ±0.4) pg/ml, t=2.683, P<0.01).In the thymus tissues of the MG patients, the mRNA levels of IL-17R were much higher than that of the controls ( relative level 2.31 folds, t =2.682, P <0.01).Meanwhile, a positive correlation was found between the plasma IL-17 levels and the relative IL-17R levels in thymus tissues ( r =0.945 4, P <0.01 ).Furthermore, IL-17 was positively correlated with quantitative myasthenia gravis scores (QMGS) either pre-treatment (r =0.798 1, P <0.01) or post-treatment (r=0.906 5, P<0.01).And IL-17R was positively correlated with QMGS pre-treatment (r=0.775 5, P<0.01).Conclusions IL-17 is increased in the plasma of MG patients (both ocular and generalized) , and is decreased upon the glucocorticoid treatment or surgery therapy, suggesting that it can be used as a parameter to determine the therapeutic effects.IL-17R is increased in the thymus tissues of MG patients, suggesting that it can potentially be used as a pathological diagnosis parameter.