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.

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.

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 explore guiding role of three-column theory in manipulative reduction, small splint fixation and early rehabilitative exercises of Colles fracture.

METHODSFrom August 2011 to February 2012,47 patients with Colles fractures were treated by manipulative reduction small splinting fixation and early rehabilitative exercises under the guidance of three-column theory, including 21 males and 26 females aged from 40 to 76 years old with an average of (65.5 +/- 2.3). According to AO fracture classification, 27 patients were type A (including 18 cases with type A2 and 9 cases with type A3) and 20 patients were type C (including 10 cases with type C1, 6 cases with type C2 and 4 cases with type C3). Pain and recovery time of swelling, postoperative complications were observed and recorded, Gartland-Werley scoring system were applied for evaluate functional recovery and biomechanical analysis of wrist joint at 12 weeks after operation.

RESULTSAll patients were followed up for 3 to 9 months with average of 5 months. Pain relief time ranged from 5 to 15 d with average of (7.6 +/- 2.2) d,recovery time of swelling of opisthenar was for 6 to 13 d with an average of (8.9 +/- 1.9) d. Two patients occurred tension vesicle within 3 days after operation, but no other complications occurred. According to Gartland-Werley scoring system, 25 cases got excellent results, 18 cases good and 4 cases moderate at 12 weeks after operation.

CONCLUSIONUnder the guidance of three-column theory, treating Colles fracture by manipulative reduction, small splinting fixation and early rehabilitative exercises can reduce pain and swelling time, promote union of fracture, effectively rehabilitate wrist function, improve clinical efficacy, and fit for concept of biomechanics.

Adult ; Aged ; Colles' Fracture ; rehabilitation ; surgery ; therapy ; Exercise Therapy ; Female ; Fracture Fixation ; Humans ; Male ; Middle Aged ; Treatment Outcome

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 investigate the blood-saving effect of controlled low central venous pressure (CLCVP) in different types of hepatectomy.Methods Ninety ASA physical status Ⅰ or Ⅱ patients of both sexes,aged 37-76 yr,weighing 40-75 kg,undergoing elective hepatectomy,were divided into 6 groups according to the surgical approach and whether CLCVP was used during surgery (n =15 each):CLCVP1-3 groups and nonCLCVP1-3 groups (NCLCVP1-3 groups).The standard hepatectomy,half liver resection and irregular hepatectomy were performed in CLCVP1-3 groups,respectively,with CLCVP.The standard hepatectomy,half liver resection and irregular hepatectomy were performed in NCLCVP1-3 groups,respectively,without CLCVP.In CLCVP1-3 groups,from skin incision to the end of liver resection,CVP was maintained ≤ 5 cm H2 O through adjustment of the position,fluid restriction and iv infusion of nitroglycerin,and norepinephrine was infused simultaneously to maintain mean arterial pressure ≥ 60 mm Hg.In NCLCVP1-3 groups CVP was maintained at 6-12 cm H2O.Intraoperative blood loss and blood transfusion were recorded.Results Compared with NCLCVP1-3 groups,intraoperative blood loss was significantly decreased in CLCVP1-3 groups (P ＜ 0.05).Compared with NCLCVP3 group,the amount of blood transfusion was significantly decreased,the constituent ratio of intraoperative blood loss ＜ 200 ml was increased,and the constituent ratio of intraoperative blood loss ＞ 1000 ml was decreased in group CLCVP3 (P ＜ 0.05).Conclusion CLCVP can decrease the intraoperative blood loss and blood transfusion in patients undergoing irregular hepatectomy.