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 investigate the hemodynamic changes of contralateral vertebral arteries with transcranial Doppler (TCD)in patients with subclavian artery steal syndrome. Methods Forty-five outpatients or inpatients with subclavian artery stenosis or occlusion confirmed by TCD and color Doppler flow imaging (CDFI)and treated at Baoan District Center Hospital of Shenzhen from March 2012 to April 2015 were analyzed retrospectively. According to the TCD detection,the degrees of intracranial arterial steal were divided into 4 groups:non-blood steal (n=8 ),blood steal phase Ⅰ(n=21 ),phase Ⅱ(n=11 ), and phase Ⅲ(n =5 );45 healthy subjects from Baoan District Central Hospital were used as a control group. The contralateral vertebral artery hemodynamic indexes of each blood steal group were detected respectively,and they were compared with the control group. Results Of the patients without blood steal,6 patients had mild subclavian artery stenosis,2 had moderate stenosis;of the patients with phase I blood steal,12 had mild subclavian artery stenosis,9 had moderate stenosis;of the patients with phaseⅡblood steal,3 had moderate subclavian artery stenosis,7 had severe stenosis,and 1 had occlusion;of the patients with phase Ⅲ blood steal,3 had severe subclavian artery stenosis,2 had occlusion. The degree of blood steal was positively correlated with the lesion degree of subclavian artery stenosis (r=0. 78, P<0. 05). TCD findings revealed that the contralateral vertebral artery systolic blood flow velocities in patients with phase Ⅰ,Ⅱ,andⅢsteal groups were 70 ± 23,85 ± 30,and 104 ± 32 cm/s,respectively;their mean flow velocities were 39 ± 10,46 ± 16,and 54 ± 17 cm/s,respectively;their pulsatility indexes were 1. 17 ± 0. 20,1. 27 ± 0. 31,and 1. 43 ± 0. 36,respectively,and they were all significantly higher than 49 ± 9,34 ± 7,and 0. 66 ± 0. 08 cm/s of the control group (all P<0. 01),and 50 ± 11,34 ± 10,and0.68±0.12cm/s of the non-blood steal group (all P<0.01),and there were significant differences among the steal blood groups in each phase (all P <0. 01 ). Conclusion Detecting the hemodynamic indexes of contralateral vertebral arteries using TCD can preliminaryly assess the subclavian artery lesions and their steal degree.

AIM:To observe the effects of Liangxue-Jiedu (LXJD) prescription, a Chinese medicine, on the expression of CC chemokine ligand 20 ( CCL20 ) and CC chemokine receptor 6 ( CCR6 ) in imiquimod-induced psoriasis-like skin lesions in a mouse model .METHODS: Male BALB/c mice were randomly divided into control group , model group, LXJD treatment group and chemokine CCL 20 monoclonal antibody treatment group .The mouse model of psoriasis was induced by imiquimod .The severity of psoriasis was assessed by the dermatologists using psoriasis area and severity in -dex (PASI).The morphological changes of the skin tissues were observed under light microscope .The thickness of epider-mis was measured .Real-time PCR was used to detect the expression of CCL 20 and CCR6 in the skin tissue samples .RE-SULTS:The skin in model group showed a large number of scales , erythema and skin thickening .Compared with model group, the skin lesion in LXJD treatment group was attenuated .The erythema and scales were alleviated , the PASI score was reduced , and the expression of CCL 20 and CCR6 was significantly lower than that in model group .CONCLUSION:LXJD prescription down-regulates the expression of CCL 20/CCR6, thus attenuating the psoriasis skin lesions in mice .

Objective To explore the expression of GPR30,HRG1 and HER2 including the activation status of HER2 (phosphorylated HER2)in invasive ductal breast cancers and their relationship with lymphatic metastasis.Methods The expres-sion of GPR30,HRG1,HER2 and pHER2 in 72 cases of specimens of invasive ductal breast cancers were examined by immunohis-tochemistry method.Results A moderate correlation between GPR30 and HRG1 was disclosed (r=0.597,P =0.000).There was strong correlation between pHER2 and GPR30 or HRG1(r=0.742,P =0.000;r=0.615,P =0.000).The expression of GPR30 and pHER2 in the lymphatic metastasis group was remarkably higher than in the group without lymphatic metastasis(P <0.05). Conclusion The interaction between GPR30 and HRG1 HER2 signal transduction pathways might be involved in the lymphatic metastasis in breast cancer.Blocking both of GPR30 and HRG1 signaling pathway could be a promising new strategy for breast cancer treatments.

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

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.

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.

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.