To prepare polyrotaxane-camptothecin conjugates and evaluate its anti-tumor effect, polyrotaxane-camptothecin conjugates were successfully synthesized, and the release behavior was performed; MTT assay and cell morphology were used to examine the inhibition of cells' proliferation effect in vitro. The experimental study of the antitumor effect on S180 mice in vivo was also performed to further evaluate the anti-tumor effect of conjugate. The result showed polyrotaxane-camptothecin conjugates can effectively inhibit the proliferation in a dose dependent effect. In vivo study and cell morphology observation of S180 mice showed significant decrease in growth of tumor, degree of tumor infiltration and blood vessel number. The result indicated anti-tumor mechanism may be through affect the angiogenesis and reduced blood supply to tumor cells and then leading to necrosis.
Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; Camptothecin ; administration & dosage ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclodextrins ; chemistry ; Drug Carriers ; Drug Compounding ; Female ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; Ovarian Neoplasms ; pathology ; Poloxamer ; chemistry ; Rotaxanes ; chemistry ; Sarcoma 180 ; pathology ; Tumor Burden ; drug effects

Objective To study the relationship between erythromyein sensitivity and ermB gene in 143 Ureaplasma urealyticum (Uu) clinical isolates. Methods We detected the minimum inhabit concen-trations (MICs) of Uu to erythromycin by broth dilution method and MIC≥8 μg/ml was used as standard concentration of resistance to erythromycin. Polymerase chain reaction was used to detect the ermB gene and biotype Uu with primers based on multi-band antigen gene. Results The MICs, MIC50 MIC90 of Uu to erythromycin were ≤0. 125 μg/ml to ≥128 μg/ml, 16 μg/ml, and ≥128 μg/ml, respectively, with a high resistance rate of 64.38%. ermB gene, which was mainly detected in Uu with MIC≥8 μg/ml, was positively detected in 40 out of 143 Uu strains (27.97%). No significant differences of the resistance to erythromycin and positive rate of ermB gene were found between the two biovars in the study . Conclusion ermB gene may probably be one of the important genes conferring resistance to erythromycin in Uu. Further studies are needed to discover the difference of resistance and mechanism of erythromycin between the two bi-ovars.

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 detect the msr gene which confers resistance to erythromycin, and ana-lyze its distributing difference between the two biovars of Ureaplasma urealyticum. Methods Broth dilution method was used to determine the minimum inhibitory concentrations (MIC) to erythromycin among 72 U. urealyticum clinical isolates. The msrA, msrB, msrC and msrD genes detection and biotyping of U. urea-lyticum were conducted using PCR. Results The MICs of 72 U. urealyticum isolates to erythromycin ranged from ≤0. 125 μg/ml to ≥128 μg/ml. MIC_(50) was 32 μg/ml and MIC_(50) was ≥128 μg/ml. Biotyping showed that biovar Parvo had 51 strains (51/72, 70.83%) and biovar T960 had 21 (21/72, 29.17%) strains.The msrA, msrB, msrC and msrD genes were obtained in 1, 12, 0 and 24 strains, respectively, with five strains carrying the msrB and msrD genes, and one strain carrying the msrA, msrB and msrD genes. There was no resistance difference to erythromycin between the two biovars when the MIC≥8 μg/ml was considered resistance to eryt hromycin. But the msrB gene was predominantly detected in biovar T960. Conclusion U. urealyticum clinical isolates harbeur the msrA, msrB and msrD genes, and the predominantly detected msrB gene is of biovar T960.

OBJECTIVETo observe the efficacy difference of electroacupuncture and auricular acupuncture in the treatment of methamphetamine withdrawal syndrome.

METHODSNinety male patients of methamphetamine addiction were randomized into an electroacupuncture group, an auricular acupuncture group and a control group, 30 cases in each one. In the electroacupuncture group, Neiguan (PC 6), Shenmen (HT 7), Zusanli (ST 36), Sanyinjiao (SP 6), Jiaji (EX-B 2) at T5 and L2 were selected bilaterally. In the auricular acupuncture group, jiaogan (AH(6a)), shenmen (TF4), fei (CO14) and gan (CO12) were selected unilaterally. The treatment was given 3 times a week, totally 12 treatments were required. In the control group, no any intervention was applied. Separately, before treatment and after 1, 2, 3 and 4 weeks treatment, the scores of methamphetamine withdrawal syndrome, Hamilton anxiety scale and Hamilton depression scale were observed in each group.

RESULTSThe total score of methamphetamine withdrawal syndrome, anxiety score and depression score were obviously reduced in 2, 3 and 4 weeks of treatment as compared with those before treatment in the electroacupuncture group and the auricular acupuncture group (all P < 0.05), and showed a trend of gradual decline as the extension of treatment. In 1,2,3,4 weeks of treatment, the total score of withdrawal syndrome, anxiety score and depression score in the electroacupuncture group and auricular acupuncture group were lower significantly than those in the control group (all P < 0.05), in which, the total score of withdrawal syndrome in the electroacupuncture group was lower significantly than that in the auricular acupuncture group in the 4th week of treatment (3.69 +/- 2.446 vs 5.73 +/- 3.169, P < 0.05); the anxiety scores were lower significantly than those in the auricular acupuncture group in 3 and 4 weeks of treatment (8.19 +/- 4.57 vs 9.65 +/- 4.24, 5.27 +/- 2.89 vs 7.38 +/- 3.10, both P < 0.05); the depression scores were lower significantly than those in the auricular acupuncture group in 2, 3 and 4 weeks of treatment (15.35 +/- 5.64 vs 19.81 +/- 5.37, 10.96 +/- 4.52 vs 15.00 +/- 4.53, 7.96 +/- 2.69 vs 12.35 +/- 3.59, all P < 0.05).

CONCLUSIONElectroacupuncture at the body points and auricular acupuncture play the therapeutic role in the treatment of methamphetamine withdrawal syndrome, anxiety and depression. The longer time the treatment is with electroacupuncture at the body points, the more obvious the efficacy will be on the above symptoms.

Acupuncture Points ; Acupuncture, Ear ; Adult ; Electroacupuncture ; Female ; Humans ; Male ; Methamphetamine ; adverse effects ; Middle Aged ; Substance Withdrawal Syndrome ; etiology ; therapy ; Treatment Outcome ; Young Adult

Objective To explore the curative effect of 308 nm excimer laser on child vitiligo and nursing measures. Method Fourty-eight children with vitiligo were treated with 308 nm excimer laser and nursed with targeted psychological treatment, skin and diet nursing.Result After treatment,6 of them got recovered(12.5%),24 remarkably improved(50%),15 slightly improved(31.2%)and 3 ineffectively treated(6.2%).Conclusions 308 nm excimer laser is effective for the treatment of child vitiligo.High quality nursing can ensure and improve the efficiency of the 308 nm excimer laser.

Objective To explore the diagnostic significance of differential cell count in induced sputum to chronic cough and assessment of airway inflammation.Methods The sputum of 335 chronic cough patients were induced.Differential cell counts were measured in these samples.The side effects were observed during the induced procedure.The final diagnosis was made based on clinical manifestation and examination findings including pulmonary function tests,provocation test,induced sputum cell differentials, etc.Results The cause of chronic cough was defined in 322 patients.The six most important causes of cough were typical asthma(TA,n=84),eosinophilic bronchitis (EB,n=62),atopic cough (AC,n= 42),cough variant asthma (CVA,n=40),gastroesophageal reflux cough(GERC,n=37),rhinitis and/ or paranasal sinusitis (PNDs,n=32),and others and indefinite cause (n=25,13).Percentage of eosinophils were significantly increased in the induced sputum of AC,EB,CVA,and GERC patients (0.005,0.052,0.059,0.234) compared with those in other causes and the healthy controls (0) (P

Objective To investigate whether ultraviolet A UVA)-induced CatK expression is regulated by the mitogen-activated protein kinases (MAPK) signaling pathway in human dermal fibroblasts in vitro.Methods Human dermal fibroblasts were obtained from circumcised foreskin of children,and subjected to primary culture.After several passages of subculture,some fibroblasts were irradiated with UVA at a dose of 10 J/cm2.Western blot was performed to measure the expressions of total and phosphorylated JNK (t-and p-JNK) and P38 (t-and p-P38) at 0.75,1.5,3 and 6 hours after the irradiation.Some fibroblasts were divided into six groups:control group receiving no treatment,SP group treated with SP600125 of 800 nmol/L,SB group treated with SB203580 of 10 μmol/L,UVA group irradiated with UVA at a dose of 10 J/cm2,UVA-SP group treated with SP600125 for 1 hour before and for 1.5 or 48 hours after UVA irradiation at 10 J/cm2,UVA-SB group treated with SB203580 for 1 hour before and for 1.5 or 48 hours after UVA radiation at 10 J/cm2.Subsequently,Western blot was performed to determine the expressions of p-c-Jun and p-MAPKAPK2 in these groups at 1.5 hours after the UVA irradiation,and reverse transcription (RT)-PCR and Western blot to detect the mRNA and protein expressions of CatK at 48 hours after the UVA irradiation,respectively.Statistical analysis was carried out by t test,one way analysis of variance and least significant difference (LSD)-t test.Results The expression levels (gray values) of p-JNK and p-P38 were significantly increased at 0.75 hour (4.77 ± 0.19 and 2.44 ± 0.13 respectively,both P ＜ 0.05) and 1.5 hours (4.68 ± 0.09 and 2.30 ± 0.04 respectively,both P ＜ 0.05),but showed no significant changes at 3 hours (both P ＞ 0.05) and 6 hours (both P ＞ 0.05) after the UVA irradiation compared with those before the irradiation (3.2 ± 0.27 and 1.61 ± 0.08 respectively).A significant decrease was observed in the expression of p-c-Jun in the UVA-SP group and p-MAPKAPK2 in the UVA-SB group compared with the UVA group (p-c-Jun,2.55 ± 0.48 vs.4.85 ±0.96; p-MAPKAPK2,1.16 ± 0.12 vs.2.46 ± 0.09,both P ＜ 0.05).The CatK mRNA and protein expressions were attenuated by 61.1％ and 44.3％ respectively in the UVA-SP group (both P ＜ 0.05),and by 71.3％ and 50.4％ respectively in the UVA-SB group (both P ＜ 0.05) in comparison with the UVA group.The UVA-SP group also showed a significant reduction in CatK mRNA and protein expressions as compared with the UVA-SB group (both P ＜ 0.05).Conclusion Both JNK and P38 signaling pathways,especially the JNK pathway,may contribute to the upregulation of CatK expression in dermal fibroblasts induced by UVA irradiation.

Objective To find the mutation of disease-causing genes of sudden unexplained death syn-drome (SU D S ) in the young by whole exome sequencing in one case. Methods O ne SU D S case was found no obvious fatal pathological changes after conventional autopsy and pathological examination. The whole exome sequencing was performed with the Ion Torrent PGMTM Systemwith hg19 as reference se-quence for sequencing data. The functions of mutations were analyzed by PhyloP, PolyPhen2 and SIFT. A three-step bioinformatics filtering procedure was carried out to identify possible significative single nu-cleotide variation (SN V ), which was missense mutation with allele frequency <1% of myocardial cell. Results Four rare suspicious pathogenic SN V were identified. C ombined with the analysis of convention-al autopsy and pathological examination, the mutation MYOM 2 (8_2054058_G/A ) was assessed as high-risk deleterious mutation by PolyPhen2 and SIFT, respectively. Conclusion Based on the second genera-tion sequencing technology, analysis of whole exome sequencing can be a newmethod for the death cause investigation of SU D S. The gene MYOM2 is a newcandidate SU D S pathogenic gene for mecha-nismresearch.

Objective To investigate the effect of ultraviolet A (UVA) radiation on the expression and secretion of cathepsin G (CatG) by human dermal fibroblasts.Methods Dermal fibroblasts were isolated from the foreskins of boys,and subjected to primary culture and subculture.After 10 or less passages,the fibroblasts were collected and divided into several groups to be irradiated with 10 J/cm2 UVA followed by 24,48 and 72 hours of additional culture,or be irradiated with 10,20 and 30 J/cm2 UVA followed by 24 hours of additional culture,with those receiving no treatment serving as the control group.Subsequently,cells and culture supernatant were collected,real time PCR and Western blot were performed to detect the expressions of CatG mRNA and protein respectively in these cells,and enzyme-linked immunosorbent assay (ELISA) was conducted to measure the expression of CatG protein in the culture supernatant of these cells.Results Compared with the control group,the fibroblasts irradiated with 10 J/cm2 UVA showed a significant increase at 24,48 and 72 hours in the expressions of CatG mRNA (0.376 ± 0.014 vs.0.183 ± 0.003,0.308 ± 0.022 vs.0.185 ± 0.005,0.296 ± 0.032 vs.0.182 ± 0.004,respectively,all P＜ 0.05) and protein (1.80 ± 0.12 vs.0.96 ± 0.06,1.41 ± 0.17 vs.0.95 ± 0.22,1.27 ± 0.09 vs.1.00 ± 0.14,respectively,all P ＜ 0.05),as well as in the supernatant level of CatG protein ((161.35 ± 7.55) vs.(122.45 ± 6.46) ng/L,(141.76 ± 2.95) vs.(124.17 ± 6.15) ng/L,(139.63 ± 3.04) vs.(121.72 ± 3.17) ng/L respectively,all P ＜0.05),with the strongest increase observed at 24 hours.At 24 hours after 10,20 and 30 J/cm2 of UVA radiation,the expression of CatG mRNA in irradiated fibroblasts was 1.90,2.51 and 3.04 times respectively (all P ＜ 0.05),the expression of CatG protein was 1.88,3.97 and 4.72 times respectively (P ＜ 0.05),and the supernatant level of CatG protein was 1.36,1.50 and 1.66 times respectively (P ＜ 0.05),that in the control group,and there was an increasing trend in all the above three parameters with increasing dose of UVA.Conclusion Acute UVA radiation can promote the expression and secretion of CatG by human dermal fibroblasts.