Porphyromonas gingivalis is a specific causative agent of human chronic periodontitis. This anaerobe produce collagenase PrtC encoded by prtC. To constructed the prokaryotic expression system for the prtC gene from P.gingivalis so as to be used to explore antigenicity and immunoreactivity of prtC gene product as well as the relationship between the local level of PrtC and the periodontitis damage, the entire length of prtC genes fragment from the ATCC-33277 and 47A-1 strains of P.gingivalis was amplified by PCR. After T-A cloning and sequencing, the prokaryotic expression system for prtC was constructed by using pET32a plasmid and E.coli BL21DE3 strain. The expression of the target recombinant PrtC protein was induce with different concentrations of IPTG. Western blot assay was used to detect the antigenicity and immunoreactivity of PrtC protein, and ELISA assay was used to detect the PrtC level in the subgingival plaque samples of patients with chronic periodontitis. The experimental results showed that the nucleotide sequences of the prtC genes from ATCC-33277 and 47A-1 strains of P.gingivalis were entirely identical, and the sequence similarities of nucleotides and amino acids were 98.46% and 99.07% respectively. Under the induction of different concentration of IPTG, the output of recombinant expressed product PrtC may reach up to 50% of the total bacterial proteins. It was also proved that the recombinant PrtC could bind with antibody against the whole cell of P.gingivalis, and could induce the production of specific antibodies in rabbit. In 91.39% of the subgingival plaque samples, the PrtC could be detectable, in which the level of positive rates of detection was higher in severe cases of chronic periodontitis than that in the mild cases. So far, a prokaryotic expression system for the prtC gene from P.gingivalis with high expression efficiency was successfully constructed in the present study, and the expressed product PrtC possesses well antigenicity and immunoreactivity, suggesting the possibility to be used as the candidate antigen for developing the serological kit and P.gingivalis vaccine.

Objective: To explore influence of bisoprolol on neuroendocrine factor levels in patients with chronic heart failure (CHF), and its therapeutic effects.Methods: A total of 116 CHF patients, who were treated in our department from Jan 2014 to Nov 2015, were selected.According to random number table, they were randomly and equally divided into routine treatment group and bisoprolol group.Left ventricular ejection fraction (LVEF), left ventricular end-diastolic dimension (LVEDd), 6min walking distance (6MWD), serum levels of brain natriuretic peptide (BNP), endothelin (ET) and atrial natriuretic peptide (ANP) before and three months after treatment, and therapeutic effect were compared between two groups.Results: There were no significant difference in above indexes between two groups before treatment, P>0.05 all.Compared with routine treatment group after three-month treatment, there were significant rise in LVEF [(28.13±4.16)% vs.(36.02±5.14)%] and 6MWD [(245.74±44.62)m vs.(315.23±53.12)m], significant reductions in LVEDd [(62.73±10.14)mm vs.(52.82±9.47)mm], serum levels of BNP [(424.51±62.26) ng/ml vs.(324.35±50.46) ng/ml], ET [(80.14±13.25) ng/ml vs.(57.91±10.32) ng/ml] and ANP [(233.24±42.35)ng/ml vs.(164.83±31.26) ng/ml] in bisoprolol group, P<0.05 all.Compared with routine treatment group, there was significant rise in total effective rate (72.4% vs.86.2%) in bisoprolol group, P=0.04.Conclusion: Bisoprolol can significantly reduce BNP, ET and ANP levels, then delay ventricular remodeling and improve cardiac function in patients with chronic heart failure.The therapeutic effect is significant, which is worth extending.

Objective To realize the risk factors for postoperative healthcare-associated infection (HAI)in pa-tients undergoing oral and maxillofacial malignant tumor surgery,so as to take effective intervention measures and reduce the occurrence of HAI.Methods Prospective and retrospective survey were adopted to analyze the occur-rence of HAI and related risk factors for postoperative HAI in patients undergoing oral and maxillofacial malignant tumor surgery in 2013.Results Of 432 patients,58 developed 63 times of HAI,HAI rate was 13.43%,case infec-tion rate was 14.58%.The main infection sites were lower respiratory tract (57.14%)and surgical site (38.09%). 56 pathogenic strains were isolated,the major were Pseudomonas aeruginosa (46.43%),followed by Acinetobacter baumannii and Klebsiella pneumonia .Univariate analysis revealed that tracheotomy,length of hospital stay,opera-tive time,intraoperative antimicrobial use were risk factors for HAI in patients undergoing oral and maxillofacial malignant tumor surgery(all P <0.05).Conclusion The important measures for reducing HAI in patients undergo-ing oral and maxillofacial malignant tumor surgery are shortening the duration of surgery and length of hospital stay, taking active intervention,implementing hand hygiene,and using antimicrobial agents rationally.

Objective To generate the spaO-ompA fusion gene of Salmonella paratyphi A and its prokaryotic expression system,and to determine the immunoprotection of the recombinant expression product rSpaO-OmpA.Methods A flexible peptide sequence was used to link spaO and ompA genes and a prokaryotic expression system of spaO-ompA fusion gene was subsequently generated.SDS-PAGE and Bio-Rad Agarose Image Analyzer were applied to examine the expression as well as the yield of the target recombinant protein rSpaO-OmpA.The antigenicity and immunoreactivity of rSpaO-OmpA were determined using immunodiffusion test,Western Blot assay and micro-Widal's test.By a mouse infection model,the immunoprotection of rSpaO-OmpA against the lethal challenge of S.paratyphi A was determined.In the animal protective test,the recombinant expressed SpaO (rSpaO) and OmpA ( rOmpA ) were used as the controls.Results The generated spaO-ompA fusion gene had 100％ nucleotide and amino acid sequence identities compared to the single spaO or ompA gene.The constructed prokaryotic expression system IPTG E.coli BL21DE3pET42a-spaO-ompA expressed the recombinant protein rSpaO-OmpA.rSpaO-OmpA combined with the antiserum against wholecell of S.paratyphi A to present positive hybridization signal and induced specific antibody in the immunized rabbits.Immunization with 100 or 200 μg rSpaO-OmpA contributed 66.7％ (8/12) or 83.3％ (10/12) immunoprotective rates in mice when the animals were attacked with S.paratyphi A.The immunoprotective rates produced by rSpaO-OmpA were significantly higher than that of equal rSpaO or rOmpA( P＜0.05 ).The sera from rSpaO-OmpA immunized mice presented 1∶5-1∶40 agglutination titers to the H antigens of different S.paratyphi species,and 1∶1-1∶16 immunodiffusion titers to rSpaO,rOmpA and rSpaO-OmpA proteins,respectively.Conclusion The artificially fusion antigen,rSpaO-OmpA,has more powerful immunogenicity and immunoprotection that the equal rSpaO or rOmpA.

Objective To investigate the effect of chemotherapy on ovarian function in young patients with malignant germ cell tumors of the ovary after conservative surgery.Methods 15 patients (11～29 years old) with ovarian malignant germ cell tumors were treated by ovarectomy and the following chemotherapy.Their menstruation and serum follicle-stimulating hormone(FSH),luteinizing hormone(LH),estradiol(E2) levels and basal antral follicle(F0) numbers were observed from the duration of chemotherapy to one year after chemotherapy.The serum levels of FSH,LH,E2 and basal antral follicle numbers between the chemotherapy-related amenorrhea(CRA) patients and the patients with benign tumors of ovary who underwent unilateral adnexa removed at the same period(control group,n=10) were compared.Results Oligomenstruation occurred in 13 of 15 cases undergoing chemotherapy after conservative surgery(86.7%),in whom CRA occurred in 10 cases(66.7%) at the duration of 4-6 cycle chemotherapy appearing significantly rising serum level of FSH,LH and declining serum E2 level and F0 numbers.There was a significantly positive correlation between the frequency of amenorrhea and the duration of chemotherapy(r=1.000,P=0.01).Generally menstruation resumed around 2～3 months after chemotherapy.Ahhough the serum levels of sexual hormone seemed to be normal at half a year after chemotherapy in CRA patients,the serum FSH(8.30±1.46 mU/ml) and E2(80.50±7.86 ng/L) level were significantly increased as compared with that of the control group and the reverse was found in Fo numbers(3.80±1.04) (t Fo,FsH,E2=7.660,5.277,7.225;P F0,FSH,E2<0.001,<0.001,<0.001).One year after chemotherapy,serum hormone level and the ovary reserve function of the CRA patient came to be normal[FSH=(5.59±0.52)mU/ml,LH=(5.25±0.84)mU/ml,E2=(56.50±5.13)ng/L,Fo=(6.50±1.08)] and that of control group [FSH=(5.43±0.35)mU/ml,LH=(5.17±0.48)mU/ml,E2=(58.10±3.73)ng/L,F0=(6.60±0.97)](t=1.177,0.694,0.505,0.740;P>0.05,>0.05,>0.05,P>0.05).Conclusions Ovarian function impairment may occur in young patients in adolescence and sexual maturity period with ovarian malignant germ cell tumors who were treated by conservative surgery followed by chemotherapy.Although the CRA is reversible,the recovery of menstruation does not meail the recovery of ovarian funetion.because the ovarian impairment may continue to a duration of half a year after chemotherapy.The drug which can safeguard ovariall function should be considered during chemotherapy.

Objective To determine the role of flagellar motor protein MotA in the pathogenesisassociated chemotaxis and colonization of Campylobacter jejuni. Methods The motA gene as well as Kan~r gene and plus-motA gene segments for motA gene knock-out were amplified by PCR and the target amplification fragments were sequenced after cloning. A suicide plasmid(pBlueskrit-Ⅱ-SK~(motA-kan)) and a motA gene knock-out mutant (motA~-) were constructed based on homologious recombination. By using semisolid plate migration test, hard agar plus (HAP)-based chemotactic test towards sodium deoxycholate (SDC) in vitro, and jejunal colonization test in BALB/c-ByJ mice were performed to determine the differences of flagellar motility, chemotaxis towards SDC and colonization in murine jejunum between motA~- mutant and wild-type strain. Results The nucleotide and amino acid sequences of the cloned motA gene were 100% identical to the reported corresponding sequences. The results of PCR, sequencing and continuous passage culture in antibiotics-contained medium demonstrated that both suicide plasmid and motA~- mutant were successfully generated. The diameters of clonies on semisolid plate and 0.2 mol/L SDC-induced chemotactic tings in HAP as well as the bacterial numbers adhering to the surface of murine jejunal mucosa and in jejunal content of motA~- mutant were significantly less than those of wild-type strain(P<0.05). Conclusion A motA gene knock-out mutant of C. jejuni was successfully constructed in this study, motA is an essential gene for flagellax motility, pathogenesis-associated chemotaxis and colonization of C. jejuni.

Objective To generate a prokaryotic expression system of Salmonella paratyphi A nmpC gene that encoding an outer membrane protein(OMP),and to determine immunogenicity and immonuprotection of the recombinant expressed product rNmpC and carrying and expression frequencies of the nmpC genes in isolates of S.paratyphi A.Methods A nmpC gene clone was obtained from a clinical S.paratyphi A strain JH01 by PCR and T-A cloning method,and then a prokaryotic expression system of the gene clone was generated.SDS-PAGE and Bio-Rad Agarose Image Pattern Analysis System were applied to examine the expression and yield of rNmpC.Antigenicity and immunoreactivity of rNmpC were determined by immunodiffusion test,Western blot assay and micro-Widal's test.The carrying and expression rates of nmpC genes in 98 S.paratyphi A isolates were detected by PCR and ELISA.By a mouse infection model,the immunoprotective effect of rNmpC against the lethal challenge of S.paratyphi A was determined.Results All the cloned nmpC genes had 100% nucleotide and putative amino acid sequence identities compared to the reported sequencing data.The expression yield of rNmpC was approximately 30% of the total bacterial proteins.rNmpC could efficiently induce rabbits to produce specific antibody and present positive Western hybridization signals with S.paratyphi A antiserum.All the tested S.paratyphi A strains have nmpC gene as well as express NmpC protein,but no nmpC gene could be detectable in S.typhi,S paratyphi B and S paratyphi C.Immunization with 100 μg and 200 μg rNmpC contributed 41.7%(5/12) and 66.7%(8/12) immunoprotective rates in mice,respectively.The sera from rNmpC immunized mice and survival mice in the NmpC is an unique OMP antigen of S.paratyphi A with conserved sequence,extensive distribution and natural expression.This OMP can be used as one the candidate antigens for developing multiple-valence genetic engineering vaccine of S.paratyphi A based on its fine immunogenicity and certain immunoprotection.

BACKGROUND: Whole blood viscosity is the keystone in hemorheological research, which has important significance in the exposure of the generation of ischemic diseases including myocardial infarction (MI) and cerebral infarction(CI) due to hemorheological changes.OBJECTIVE: To investigate the characters of macroscopical and microscopical hemorheology to explore the standardization of hemorheological measuring method.DESIGN: An observatory comparative study based on the venous blood of healthy individual.SETTING: Department of physics in a medical college.MATERIALS: Venous bloods of 86 healthy individuals were randomly selected for the study, which was conducted in the Laboratory of Hemorheology of Guilin Medical College.METHODS: Totally 86 whole blood samples were tested under 7 shear rates (0.6 s-1,2 s-1,5 s-1, 10 s-1, 20 s-1, 40 s-1, 100 s-1) from low shear rate to high shear rate, and then from high shear rate to low shear rate. The measuring value of the two processes was compared.Casson viscosity and yield stress in two measuring process.RESULTS: There was no significant difference of apparent viscosity( P＞ 0.05 ) except the 0.6 s- 1 -corresponding apparent viscosity ( P ＜ 0.05 )at some shear rate in two measuring process. There was no significant difference in Casson viscosity and yield stress( P ＞ 0.5, P ＞ 0. 3).CONCLUSION: The measurement of Casson viscosity and yield stress is good for the optimization of the standardization of hemorheological indicators and measuring method; and it is feasible to test apparent viscosity, Casson viscosity and yield stress repeatedly in same blood sample, which has important significance in the exposure of the hemorheological changes and the generation and development mechanism of ischemic diseases including MI,CI, hypertension, and coronary heart disease(CHD).

Antimetabolites, Antineoplastic ; pharmacology ; Apoptosis ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; Female ; Fluorouracil ; pharmacology ; Gene Expression Regulation, Neoplastic ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; physiology ; Neoplasm Proteins ; biosynthesis ; genetics ; physiology ; Oligodeoxyribonucleotides, Antisense ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Transfection

BACKGROUND:At present, a large number of studies have proved that the expandable intramedul ary nail and interlocking intramedul ary nail are effective in treatment of limb fractures. However, for the treatment of tibial fractures, the advantages and disadvantages of the two kinds of fixation methods are stil inconclusive. OBJECTIVE:To systematical y review the effectiveness and safety of expandable intramedul ary nail and interlocking intramedul ary nail for treatment of tibia fracture. METHODS:We searched PubMed, Embase, The Cochrane Library (Issue 1, 2015), CBM, CNKI, VIP and WanFang Data for articles concerning randomized control ed trials on expandable intramedul ary nail versus interlocking intramedul ary nail published from inception to January 1, 2015. The key words were“tibia, fracture, tibia fracture, tibia fractures, expandable intramedul ary nail, expandable nail, intramedul ary nail, interlocking intramedul ary nail”. Meta-analysis was performed using RevMan 5.2 software. RESULTS AND CONCLUSION:Ten randomized control ed trials involving 574 patients were included. The results of meta-analysis showed that, compared with interlocking intramedul ary nail, expandable intramedul ary nail was shorter in operation time [MD=-23.42, 95%CI (-26.94,-19.90), P<0.000 01], less in intraoperative hemorrhage [MD=-47.64, 95%CI (-52.21,-43.09), P<0.000 01], less in fluoroscopy times [MD=-1.40, 95%CI (-1.49,-1.30), P<0.000 01], shorter in union time [MD=-30.84, 95%CI (-35.27,-26.41), P<0.000 01], and less in incidence of complications [OR=0.20, 95%CI (0.10, 0.40), P<0.000 01]. The Johner-Wruh scores showed on significant difference. These findings suggest that expandable intramedul ary nail for tibia fracture has more advantages than interlocking intramedul ary nail. That is to say, the expandable intramedul ary nail is an improved nail of interlocking intramedul ary nail, but a large sample of high quality randomized control ed trials are stil needed to confirm the conclusion.