Animals ; Gene Silencing ; physiology ; Gene Targeting ; Genetic Therapy ; methods ; Genomics ; Humans ; RNA Interference ; RNA, Small Interfering ; genetics ; therapeutic use ; RNA-Induced Silencing Complex

AIM: To study the possible role of nitric oxide (NO) in the development of periodontitis and the relationship between the NO concentration and the attachment loss. METHODS: Seventy- two Sprngue- Dawley rats were randomly assigned to two groups, the control group and periodontitis group. Experimental periodontitis in rats was produced by a ligature of braided silk. The nitric oxide concentration was indirectly ascertained by the concentration of nitrite (NO2-) and nitrate (NO3-)in the gingival tissue, which was assayed by spectrophotometry. The attachment loss (AL) was measured by the technology of the cellular graphics engineering research. The histopathologic change in periodontium was observed under a light microscope by using the histotomy. RESULTS: Compared to control group, the NO2-/NO3 - concentration in gingival tissue was significantly higher in periodontitis group at four weeks and eight weeks following ligation (P＜0.01). In periodontitis group, the NO2-/NO3 - concentration in gingival tissue was higher at eight weeks than that at four weeks following ligation (P＜0.01). At four weeks and eight weeks, the AL in experimental periodontitis in rats was significantly increased than that at one week after ligation ( P＜0.01); and the AL was also much higher at eight weeks than that at four weeks (P＜0.01). CONCLUSIONS: The NO2-/NO3- concentration in the gingival tissue in periodontitis group was significantly higher than that in control group. These results demonstrate that the NO2-/NO3- concentration is related to the severity of AL, and NO synthesis is very important to the process of inflammation and lesion in periodontium. Reducing NO production may be of great therapeutic value in the treatment of periodontitis.

Objective To introduce an interpersonal relationship regulating technique in the static state of Qigong to enrich modern clinical psychotherapies. Methods Based on previous therapeutic experience, we introduce one specific technique of the Systematic Psychotherapy of TCM (SPT): Interpersonal relationship regulating technique in the static state of Qigong. Results Interpersonal relationship regulating technique in the static state of Qigong is a specific and practical technique of SPT, which has been gradually standardized and been mature with more than two decades of innovation and accumulation. Conclusion Interpersonal relationship regulating technique in the static state of Qigong has great clinical value.

Objective:To survey the status quo of ADR occurred in our hospital. Methods:Totally 277 ADR reports were collect-ed and analyzed statistically in our hospital from January 2010 to June 2013. Results:Among the ADR cases,the ratio of male to fe-male was 1∶1. 565, and 59. 93% of ADR occurred in the people over 50 years old. Totally 240 ADR cases (86. 64%) were induced by intravenous administration and 32 ADR cases (11. 87%) were induced by oral administration. Totally 167 ADR cases were caused by antibiotics (60. 29%), among them, 54 cases wero by cephalosporins (32. 34%), 47 cases by quinolones (28. 14%) and 32 ca-ses by cephamycins (19. 16%). Totally 42 ADR cases were caused by traditional Chinese medicine injections (15. 16%), and 18 ca-ses were caused by cardiovascular system drugs (6. 5%). The main clinical manifestations were lesion of skin and its appendant organr and digestive system injury with 162 cases (58. 48%) and 62 cases (22. 38%), respectively. Conclusion: It is necessary to strengthen clinical ADR monitoring, rationally use antibiotics and emphasize the importance of safe medication of traditional Chinese medicine injections.

Objective To investigate the effect of electron transfer system on the hyphal formation of Candida albicans. Methods Candida albicans was cultured in RPMI 1640 supplemented with 10% new-born calf serum in 5% CO2 at 37 ℃ with or without the presence of inhibitors or activators of electron transfer system. Growth curve, morphology and percent of filamentation were observed for Candida albicans. MTT assay was used to assess the viability of Candida albicans. Results The solvents (chloroform and dimethyl sulfoxide) had no significant effect on the growth of and filamentation in Candida albicans. After incubation with thenoyltrifluoroacetone (TTFA) or benzhydroxamic acid for 24 hours, yeast cells of Candida albicans predominated in the culture. The growth of Candida albicans was significantly inhibited in log phase by the incubation with classic respiratory chain inhibitors such as rotenone, antimycin A, oligomycin, sodium azide, TTFA and sodium malonate, compared with the controls (all P < 0.01). Benzhydroxamic acid, an inhibitor of alternative oxidative pathway, also significantly inhibited the growth of Candida albicans in log phase (t = 10.92, P < 0.01). After incubation with rotenone, antimycin A, oligomycin, sodium azide, TTFA, sodium malonate, benzhydroxamic acid and disodium gnanylate, the percentage of filamentation in Candida albicans at 12 hours was 87.49 ± 0.52, 48.75 ± 4.44, 50.33 ± 8.50, 99.00 ± 1.00, 1.60 ± 0.53, 94.01 ± 0.99, 0.00 ± 0.00 and 92.33 ± 2.08, respectively, and the growth of Candida albicans at 7 hours was inhibited by (1.34 ± 0.15)%, (70.61 ± 1.02)%, (50.63 ± 5.38)%, (17.80 ± 7.89)%, (45.17 ± 1.27)%, (10.75 ± 3.62)%, (72.46 ± 1.14)% and -(5.96 ± 4.07)%, respectively. Conclusions Hyphal formation of Candida albicans could be suppressed by inhibitors of classic respiratory chain or alternative oxidative pathway, and is mainly regulated by alternative oxidative pathway.

ObjectiveTo determine whether aadA2 gene can be translated from the ATG triplet,which there was no plausible ribosome binding site preceding it,and synthetized a functional protein in class 1 integron.MethodsSite-specific mutagenesis was used to construct aadA2 gene cassette with different start codons,together with their upstreamed promoters of variable regions were cloned into plasmid pACYC184 respective.The constructed plasmids were then transfored into Escherichia coli JM109,Western blot was used to detect the translation products of aadA2 gene with different start codons.Broth microdilution method was used to detect the minimal inhibitory concentrations to streptomycin in Escherichia coli JM109 containing aadA2 gene with different start codons.ResultsaadA2 gene can initiate translation from both ATG and GTG triplets in aminoacyl -3-adenylyltransferase protein synthesis,though there was no plausible ribosome binding site preceding the ATG triplet.Besides GTG and ATG triplets,there was other start codon downstream of the GTG triplet in aadA2 gene.The translated products that initiated from the start codons that described above were all functional AAD(3) proteins that can be detected by anti- aminoacyl -3-adenylyltransferase polyclonal antisera in Western blot and conferred different resistance levels to streptomycin in E.coli.ConclusionWhen inserted as the first gene cassette in class 1 integron,aadA2 gene can initiate translation from ATG triplet and synthetized a functional protein,though there was no plausible ribosome binding site preceding it.This structural characterization of class 1 integron can initiate translation of the open reading frame harbored in gene cassette that integrated into class 1 integron,though there was no plausible RBS preceding the start codon.This make class 1 integron be more convenience to express the genes that capture from environment.

ObjectiveTo prepare antiserum specific to aminoacy1-3″-adenylyltransferase [ AAD (3″) ],and to explore the application value of the prepared antiserum in detecting the expression levels of aadA2 gene that downstream of 8 different promoters (PcS,PcH2,PcH1,PcW,PcS-P2,PcH2-P2,PcH1P2 and PcW-P2 ) of variable regions in class 1 integron.MethodsaadA2 gene was amplified by polymerase chain reaction(PCR) and cloned into the expression plasmid pET19b.After inducing,the recombined aminoacy1-3″-adenylyltransferase[ AAD(3″)] with His-tag was expressed,purified and immunized rabbits to get anti- AAD(3″) specific serum.The prepared antiserum was used to detect the translation levels of aadA2 gene that downstream of different promoters of variable regions in class 1 integron by Western blotting (WB).Broth microdilution method was used to detect the minimal inhibitory concentrations (MIC) to streptomycin in Escherichia coli JM109 with aadA2 gene downstream of different promoters of variable regions.ResultsRecombined AAD (3″) expression plasmid pET19b-aadA2 was constructed successfully and was verified by sequence analysis.After transformed into E.coli BL21 ( DE3 ),a resoluble recombined AAD(3″) high expression strain was obtained.After fermentation,recombined AAD(3″) was purified and immunized rabbits.The anti- AAD(3″) specific serum was obtained with titer ＞ 1∶100 000.WB was used to detect the expression levels of AAD (3″),the translation product of aadA2 gene,that downstream of 8 different promoters of variable regions.The relative expression level of AAD (3″) that downstream of PcW was assumed to be 1,then the relative expression levels of AAD(3″),which all were detected 3 times independently,that downstream of PcS,PcH2,PcH1,PcS-P2,PcH2-P2,PcH1-P2 and PcW-P2 were 12.9±2.3,9.1±1.0,2.0±0.4,16.0±1.3,14.1 ±1.3,10.5±0.7 and 8.9 ±1.7 respective.Very different expression levels of AAD (3″) that downstream of different promoters of variable regions were obtained( F =32.421,P ＜ 0.01 ).The mean values of MIC,which all were detected 3 times independently,to streptomycin in E.coli JM109 with aadA2 gene downstream of PcS,PcH2,PcH1,PcW,PcS-P2,PcH2P2,PcH1-P2 and PcW-P2 were 256,256,64,128,32,128,4 and 64 mg/L respective.These results indicated the different expression levels of aadA2 gene that downstream of different promoters of variable regions can confer their host bacteria different resistance levels to streptomycin.Conclusions Resoluble recombined AAD(3″) is purified successfully and high titer anti- AAD(3″) specific antiserum is obtained from the immunized rabbits.This laid foundation for further investigation on the correlationship between the expressions of intI1 gene and the gene cassettes within variable regions.The expression levels of antibiotic gene cassettes that downstream of different promoters of variable regions are very different,so are the very different antibiotic resistance levels of the host bacteria.Therefore more attentions should be paid to the researches on the classification of promoters of variable regions when molecular epidemiology studies on the class 1 integrons in clinical isolates were conducted.

AIM To study the protective effects of Jingqi Capsules (Cordycps Militaris,Rehmanniae Radix Praeparata,Lycii Fructus,) on the damage to rat reproductive system induced by cyclophosphamide.METHODS After low and high dose groups of Jingqi Capsules and positive group were administered for two weeks in advance,cyclophosphamide [25 mg/(kg · d)] modeling was conducted consecutively by intraperitoneal injection for five days except the normal group,while the treatment groups were administered consecutively.All the rats were executed the day after the end of the modeling.Serum testosterone content was measured by radioimmunoassay;body weight,testis,epididymis and prostate of rats were weighed and organ coefficient was calculated;morphology of testis was observed by HE;sperm count and sperm motility were detected by CASA;apoptosis of cells was observed by TUNEL.RESULTS Compared with the model group,each dose group of Jingqi Capsules increased in the level of serum testosterone and organ coefficient of testis,epididymis and prostate.The damage to seminiferous epithelium relieved and the number of all spermatogenic cells increased.Sperm count and sperm motility increased.Decrease in apoptosis of cells was demonstrated by TUNEL.CONCLUSION Jingqi Capsules has protective effects on damage to rat reproductive system induced by cyclophosphamide.

Will-powered technique training in the lower resistant state is an important therapy method of TCM psychology. The therapy refers to five aspects of improving cognitive and emotional function by using TIP technique, making imaginary training in the lowered resistant state, strengthening will-power training with situation and image technique, adopting information module imprinted technique and developing training technique. The content includes cultivating abilities in independence, resolution, firmness, and self-control. The technique has application value in clinic.

Objective To study the comprehensive quality of pre-hospital care of patients with acute coronary syndrome (ACS) in our country,and the factors affecting the efficiency of medical intervention of pre-hospital care.Methods An epidemiological investigation with target population-based cross-sectional study was carried out,and specifically designed questionnaire for determining the efficiency of pre-hospital care of patients with ACS was used to collect information of 272 patients from 13 emergency pre-hospital care centers.Results After analysis of the data from 13 majormedical centers,it could be classified into 4 patterns of pre-hospital care,namely directive pattern,pre-hospital pattern,dependent pattern and independent pattern.The results of analysis showed that the distance for ambulance driving in directive pattern was shorter than that in dependent pattern,but the call-response in directive pattern was quicker than that in pre-hospital pattern and dependent pattern,while the call-arrival time in directive pattern was longer than that in independent pattern and dependent pattern.The analysis of the on-site preliminary treatment in four patterns of pre-hospital care showed that there were differences in the establishment of intravenous access,aspirin,nitrate and ECG ( P ＜ 0.05 ).The total effective rate of pre-hospital care for ACS was 48％,while effective rate of independent pattern reached 70％.Factors affecting therapeutic effect on ACS resulting in better outcomes included pre-hospital pattern ( OR =4.097 ),directive pattern ( OR =5.158 ),and use of nitrate ( OR =3.045 ) and oral administration of medicine ( OR =8.215 ). Conclusions Independent pre-hospital pattern had the best therapeutic effect in the pre-hospital care of patients with ACS,and pre-hospital pattern had the best on-site preliminary disposal.Use of nitrate and oral administration of medicine showed the best benefit to the therapeutic effect.