Objective To evaluate the effect of one-visit endodontic treatment for infected root canals with combination of nickel-titanium instruments and ultrasonic technique and warm vertical technique.Methods According to treatment,90 teeth were randomly divided into experimental and control groups.Two groups were used nickel titarlium root canal preparation.Experimental group:51 teeth for ultrasonic root canal irrigation,and warm vertical technique was employed to fill the root canal.Control group:39 teeth for rinse syringes,and warm vertical technique was employed to fill the root canal.Washing fluid was 2％ sodium hypochlorite.The root canal filling was observed after a week and one year of postoperative pain response.Results A week later,two groups of post-operative pain showed that there was significant difference in responses (x2 =15.525,P ＜ 0.05).One year later,the two groups showed no significant difference in efficacy(x2 =0.085,P ＞ 0.05).Conclusion The combination of nickel-titanium instruments and ultrasonic irrigation technique and warm vertical obturation,for the treatment of infected root canals was a method to complete the root canal filling,could reduce both the number of patients subsquent visits and the postoperative pain response,while it could improve the quality of root canal filling.

AIM: To evaluate the use of 23G minimally invasive vitreous cutting system in the first stage of anterior segment reconstruction on severe ocular injury.
METHODS: Fifteen patients with ocular injuries including the corneal and scleral rupture associated hyphema, vitreous hernia in anterior chamber, traumatic lens rupture, lens subluxation, applied 23G minimally invasive vitreous cutting system in the first stage of anterior segment reconstruction.
RESULTS: The anterior ocular media of early postoperative became transparent quickly. No exudative lemma and no complications were found relate to puncture incisions after operations.
CONCLUSION:23G minimally invasive vitreous cutting system can be used in the first stage of anterior segment reconstruction on sever ocular injury. It can reduce iatrogenic injury, operative complications and inflammatory response, also can shorten therapeutic time and create favorable conditions.

Abstract? AIM: Through the establishment of penetrating keratoplasty model of rats, to detect the role and its mechanism of immature dendritic cells with IL-10 gene modified.? METHODS: Allogeneic penetrating corneal transplantation in rat model was performed. SD rats were randomly divided into positive control group, GFP-DC group, 8-DC and IL-10-GFP-DC group.At 3d before keratoplasty, the rats were given tail intravenous injection with the same amount of PBS, bone marrow 8-DC ( DC had cultured for 8d ) from donor Wistar rats, GFP-DC after 48h transfection and IL-10-GFP-DC.Rats were observed under slit-lamp for corneal graft cases every day, and recorded rejection index and corneal graft survival time.At 14d after keratoplasty, pathologic and immunohistochemical examinations were performed.?RESULTS:Compared with GFP-DC group and 8-DC group, corneal graft survival time of IL-10-GFP-DC group was significantly longer ( P<0.01 ); at 14d after keratoplasty, corneal opacity, edema, neovascularization and rejection index of IL-10 -GFP-DC group were significantly lower (P<0.01).Pathological examination showed that in the three experimental groups corneal inflammation was lighter than the positive control group without significant central graft neovascularization. Immunohistochemistry showed: compared to the positive control group, GFP-DC group and 8-DC group, CD4+, CD8+, CD25+, IL-2+, NK+and NF-κB+positive cells in IL-10-GFP-DC group were lower(P<0.01).? CONCLUSION: After donor -derived immature dendritic cells pretreated, corneal graft survival was significantly prolonged, successfully induced corneal transplantation tolerance. CD4+, CD8+, CD25+, IL-2+, NK+and NF-κB+positive cells are involved in corneal allograft rejection regulation, IL-10-GFP-DC may reduce CD4+, CD8+, CD25+, IL-2+, NK+and NF-κB+positive cell infiltration, inhibit corneal transplant rejection.

Objective To investigate the survival status of liver cirrhosis patients without upper gastrointestinal hemorrhage after transjugular intrahepatic portosystemic shunt (TIPS).Methods From 2004 to 2013,15 liver cirrhosis patients without upper gastrointestinal hemorrhage volunteered received TIPS treatment were followed up to find out the difficulty and the success rate of TIPS procedure,the incidence of hepatic encephalopathy,upper gastrointestinal hemorrhage and improving of hypersplenism.Results The success rate of operation was 100％.The average of operation time was 60 minutes.During follow-up,no stent angulation occurred,no gastrointestinal hemorrhage happened and no one died in all 15 patients after TIPS operation.There were four patients with hepatic encephalopathy in eight weeks after operation.The anemia of four patients improved compared with that before operation.Conclusions TIPS is a safe and effective threapy in the prevention of gastrointestinal hemorrhage in the patients with liver cirrhosis accompanied with severe gastroesophageal varices.It may become the primary prophylaxis for liver cirrhosis patients without upper gastrointestinal hemorrhage.

Objective To investigate the effectiveness of endoscopy and enteral nutrition (EN) in the treatment of acute biliary pancreatitis (ABP) in elderly patients and its impact on prognosis.Methods Eighty-four elderly patients with ABP were randomly divided into two groups,including 42 cases of combination treatment group,42 cases of routine treatment group.The routine treatment group was managed with normal comprehensive treatment,and in combination treatment group,patients were treated with additional endoscopic retrograde cholangiopancreatography (ERCP),endoscopic sphincterotomy (EST) and endoscopic naso-biliary (ENBD) drainage and EN.The clinical symptoms,serum inflammatory cytokines,complications and prognosis were compared.Results The CRP,TNF-α,IL-2,and serum amylase,glucose,AST after treatment were significantly lowered than those before treatment (P ＜ 0.05).In addition,the levels of CRP,TNF-α,IL-2,and serum amylase,glucose,AST,ALB in combination treatment group were significantly lower than those in routine treatment group (P ＜ 0.05).The time to abdominal pain cessation,time to fever cessation,hospital stay in combination treatment group were (4.6± 1.3),(3.7± 1.3),(13.5± 2.7) d,which were significantly lower than those in routine treatment group [(6.1 ± 1.3),(5.5 ± 2.1),(18.1 ± 3.8) d,P ＜ 0.05].The APACHE Ⅱ score of combination group after treatment was (7.l ± 1.8) point,which was significantly lower than that in routine treatment group [(9.2 ± 2.6) point,t =3.41,P ＜ 0.05)].The incidence rates of ARF,ARDS,peri-pancreatic infection and abdominal hemorrhage,in combination group were 7.1％,4.8％,4.8％,4.8％,respectively,which were significantly lower than those in routine treatment group (23.8％,19.1％,21.4 ％,19.1％,P＜0.05).The mortality in combiration group was 4.8％,which was lower than 16.7％ in routine treatment group with no statistical significant.Conclusions It is a safe and effective to treat elderly ABP patients with endoscopy combined with EN,which can significantly reduce complications and mortality.

Objective To develop a one-step PCR assay for rapid discrimination of six Brucella species and some intraspecific biovars.Methods Using 6 pairs of primers in one-step PCR to differentiate six classical Brucella species and some biovar in ordinary PCR instrument.The tested strains including 27 reference strains of six Brucella species and 239 Brucella strains were estimated by the PCR assay and biological identification methods.Results The six Brucella species could be precisely differentiated by the onestep PCR assay from the tested strains.Five biovars and vaccine strain of B.suis species could be determined,and biovars 1,3,4 and biovars 2,5,6,7,9 of B.abortus species could be identified at the level of their biovar,moreover,biovars 1,2 and 3,and vaccine strain Rev 1 of B.melitensis species were also discriminated at the biovar and strain level.The accurate rates of the biological identification method and the PCR assay were 98.33％ and 100％ respectively.Conclusion One-step PCR assay was a rapid,specific,and low cost method for identification of Brucella species and discriminating biovars in ordinary PCR instrument.

To understand the structure of the soluble region of Enterovirus 68 3A protein, we construct a prokaryotic expression vector expressing the soluble region of EV-D68 3A protein, and identify the forms of expression product after purification. The EV-D68 3A(1-61) gene was amplified by PCR and then cloned into the expression vector pET-28a-His-SUMO. The recombinant plasmid was transformed into Escherichia coli BL21 induced by IPTG to express the fusion protein His-SUMO-3A(1-61). The recombinant protein was purified by Ni-NTA Agarose and cleaved by ULP Protease to remove His-SUMO tag. After that, the target protein 3A(1-61) was purified by a series of purification methods such as Ni-NTA, anion exchange chromatography and gel filtration chromato- graphy. Chemical cross-linking reaction assay was taken to determine the multiple polymerization state of the 3A soluble region. A prokaryotic expression vector pET28a-His-SUMO-3A(1-61) expressing the solution region of EV-D68 3A was successfully constructed and plenty of highly pure target proteins were obtained by multiple purification steps . The total protein amount was about 5 mg obtained from 1L Escherichia coli BL21 with purity > 95%. At the same time, those results determined the homomultimer form of soluble 3A construct. These data demonstrated that the expression and purification system of the soluble region of 3A were successfully set up and provide some basic konwledge for the research about 3A crystal structure and the development of antiviral drugs targeted at 3A to block viral replication.
Amino Acid Sequence ; Enterovirus D, Human ; chemistry ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Models, Molecular ; Molecular Sequence Data ; Protein Structure, Tertiary ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics ; metabolism

OBJECTIVE:To shorten time consuming for dispensing mode of automatic medicine packaging machine(ATC), and improve the efficiency of ATC. METHODS:In the inpatient pharmacy of our hospital,previous dispensing mode that two ATC matched with one tablet case separately was replaced by new mode,i.e. one ATC(another ATC was on standby)matched with two tablet cases(one of them added medicine beforehand),and the information of drug-position coordinate was equipped. The time con-sumed,average daily number of packages,financial expenses and labor costs were recorded and analyzed to evaluated the effect of new mode. RESULTS & CONCLUSIONS:Compared with previous mode,new dispensing mode improve the efficiency of ATC greatly. In previous mode,two ATCs and four pharmacists work simultaneously and spend 4.5 hours for 3 850 bags of medicine, which cost 4 400 000 yuan;in new mode,only one ATC and three pharmacists work simultaneously,and spend 3.0 hours for 4 015 bags of medicine,which cost 2 350 000 yuan. Therefore,double-universal tablet cases greatly improves the efficiency of ATC and reduce the financial expenses as machine maintenance and labor cost.

Objective To collect the families with generalized epilepsy with febrile seizures plus(GEFS+) and analyze the clinical status and heredity features of Chinese GEFS+.Voltaged-gated sodium channel ?1 subunit(SCN1B) gene of 2 families were detected,and expect to find new mutation sites.Methods All participant in the study of 2 families members were informed of voluntary participate in this investigation,health examination and blood sampling.All 6 gene exons of proband,patients and healthy control group were sequenced.The sequencing result was compared and analyzed with the normal sequence of genomic exon fragment and exon fragment sequencing result of control group through internet(BLAST).Results 1.A new G/A heterozygous polymorphism (G181A)was found in the 181th basyl of SCN1B gene exon 3,and codon was changed from TCG to TCA,both encoding serine (Ser,S).It was synonymous mutation.2.A new G/A heterozygous polymophism(G15A)was found in the 15th basyl of SCN1B gene exon 3,and codon was changed from GAG to GAA,both encoding glutamic acid(Glu,E).It belonged to synonymous mutation.3.A new T/C heterozygous polymorphism (T37C)was found in the 37th basyl of SCN1B gene exon 6.The patients genetype were:5 cases with T/C heterozygote,3 cases with T/T homozygote,2 cases with C/C homozygote.Healthy control group were all T/T homozygote.Allele frequency distribution for T was 55.0%,and 45.0% for C.4.A new A/C heterozygous polymorphism (A81C)was found in the 81th basyl of SCN1B gene exon 6.The patients genetype were:5 cases with A/C heterozygote,3 cases with A/A homozygote,2 cases with C/C homozygote.Healthy control group were all A/A homozygote.Allele frequency distribution for A was 55.0%,and 45.0% for C.Conclusions Two new heterozygous polymorphism (G181A),(G15A) were found in SCN1B gene exon 3.Two new heterozygous polymorphism (T37C),(A81C) were found in SCN1B gene exon 6.These 4 polymorphism enriched single nucleotide polymorphism(SPN) database and provided candidate sites for the research of epilepsy susceptbility polymorphisms.

Generalized epilepsy with febrile seizures plus(GEFS+) is a new epilepsy syndrome proposed by International League Against Epilepsy.At present,the progress of genetic studies of GEFS+ focus on gene mapping based on family analysis,many researches indicate that GEFS+ is associated with voltaged-gated sodium channel(SCN) gene mutation.This paper intends to discuss the relationship beween GEFS+ and SCN1B,SCN2B,SCN1A,SCN2A genes,mutations in order to improve the cognition about GEFS+.