Objective Antimicrobial peptides are the focus of recent research in oral microbiology .This study aimed to eval-uate the activity of a novel antimicrobial peptide pm 11 against oral microorganisms and its action mechanisms . Methods We ana-lyzed the effect of pm11 on oral microorganisms and determined its antimicrobial activity in the saliva environment by measuring its min -imal inhibitory concentration (MIC), minimal bactericide concentration (MBC), and bactericidal kinetics.We observed its bacteri-cidal activity on the biofilms of streptococcus mutans by confocal laser scanning microscopy (CLSM) and the structural changes in the bacterial membrane by scanning electron microscopy (SEM). Results The antimicrobial activity of pm11 varied greatly against dif-ferent oral microorganisms , with its MIC values ranging from 2 μg/mL to 256 μg/mL and its MBC values from 2 μg/mL to >256μg/mL.The bactericidal kinetics showed a decreasing survival rate of bacteria with the lengthening of the intervention time .The inhib-itory-zone diameters exhibited no significant indifference between the water solution and the sterile saliva solution .CLSM revealed an increased number of dead bacteria in the pm 11-treated biofilms , while SEM manifested obvious changes in the shape of the bacteria membrane treated with pm11. Conclusion Our findings suggest that pm11 has a broad spectrum of antimicrobial activities on oral mi-croorganisms and a potential value of clinical application .

Objective:To observe the expression of GATA-3in asthmatic mice lungs and to explore the feasibility of in-terleukin(IL-12)blockading GATA-3expression in treatment of bronchial asthma.Methods:C57BL/6mice were randomly divided into3groups:control group(group A),asthmatic model group(group B)and IL-12injection group(group C).Asth-matic models were established in group B and C.Normal saline(0.1ml)and IL-12(1?g)were injected in group B and C re-spectively on the1,3,7,9,25,26,27and28d.Six mice from each group were obtained for analyses of bronchoalveolar lavage fluid(BALF)on d31.The airway pathology changes and the expression of GATA-3were observed by hematoxylin/eosin(H-E)and immunohistochemistry respectively.Results:There was no symptom in group A,and the symptoms of group B were more severe than those of group C.Eosinophil(EOS)was not seen in the BALF of group A,while EOS in group B and group C were(20.0?4.0)%and(0.2?0.1)%respectively.In the same microscopic visual field,there was no inflam-mation cell in group A and a large number of inflammation cells in group B,but inflammation cells in group C were signifi-cantly decreased.Immunohistochemistry showed that the expression of GATA-3in group B was strong and no GATA expres-sion was found in group A and group C.Conclusion:The expression of GATA-3in asthmatic mice is high;IL-12can inhibit asthmatic airway and lungs inflammtion,whose mechanism may be the blockade of GATA-3expression in asthmatic models.[

This paper reported a new home-made nonpoisonous substrate of horseradish peroxidase(HRP),3,3′,5,5′-tetramethylbenzidine sulphate(TMBS),used in ELISA with the aqueous soluble egg antigen(SEA)as antigen;proved patients'sera as antibo- dy and the antihuman IgG was replaced by protein A conjugate. The optimun condition obtained by orthogonal design was 1.0 millimolarity of TMBS,1.2 millimolarity of H_2O_2,and pH value at 5.0.The best time to stop reaction with H_2SO_4,was 30-60 min after adding substrate TMBS.At the same time,TMB and OPD were used as compared substrates.It showed that the reaction colour of both TMBS and TMB remained much more stable than that of OPD when being exposured in light and oxygen.After the solutions of TMBS and TMB were stored under 4℃ in refrigerator for 60 days,the coefficient of variation among different experiments was less than 10%. The results in detecting specific antibody in sera from 28 patients with proved schistosomiasis also showed that the sensitivity of TMBS was remarkably higher than OPD.It was found that TMBS used in enzyme immunoassay for demonstrating specific antibody was suitable not only for optical density(OD)read by spectrophotometer but also for estimation judged by naked-eye.During TMBS and TMB used in indirect immunoperoxidase method(IIP),the specific antibody in the tissue can be easily dete- cted and located as well.

Objective To observe the induction of tolerogenic dendritic cells (DCs) by curcumin (Cur) and its mechanism.Methods After immature DCs from bone marrow cells of Wistar rats were treated with different concentrations of Cur (0,10,20 and 30?mol/L) respectively,and then the DCs were tested by flow cytometry for the surface molecules expression.After the immature DCs were treated by 30?mol/L Cur with or without stimulation of LPS,endocytosis of DCs to dextran was tested by flow cytometry.The production of IL-12 in DC culture supernatant was determined by ELISA.The levels of NF-?B p65 and RelB translocation to the nucleus were investigated by Western- blot.The activity of NF-?B was detected by NF-?B-binding ELISA and luciferase reporter gene analy- sis.The ability of DCs to stimulate the proliferation of T cells from Lewis rats were analyzed by mixed leukocyte reactions (MLR).Results Cur suppressed LPS-indueed cell-surface expression of costimu- latory molecules (CD80,CD86 and CD40) in a dose-dependent manner.When Cur was used at a con- centration of 30?mol/L,there was no marked difference in the surface molecules expression of LPS- inducing DCs as compared with immature DCs.After DCs were induced by LPS (LPS group),the positive rate of FITC-Dextran uptake was (36.6?7.2)%,and the secretory amounts of IL-12 were (415.9?42.7) pg/ml.In DCs of LPS group,the intranuclear RelB and p65 were highly expressed and their DNA binding activity was 0.65?0.08 and 0.74?0.07 respectively.The luciferase activity of reporter gene in LPS group DCs was remarkably increased to 435% as compared with that in the controls.DCs in LPS group showed strong capacity to stimulate T cells proliferation.When DCs were treated with 30?mol/L Cur followed by induction with LPS (Cur+LPS group),the positive rate of Dextran uptake was (78.6?14.2)% and remarkably higher than in LPS group (P

Objective： This study was designed to investigate the relationship between p16 protein expression and gastric carcinogenesis,depth of invasion, lymph node metastasis, and evaluate the role of deletion and mutation of p16 gene in exon 2 in gastric carcinoma. Methods： p16 protein expression in gastric carcinoma and precancerous lesion was examined by streptavidin-peroxidase conjugated(S-P) method; The deletion and mutation of p16 gene were examined respectively by polymerase chain reaction(PCR) and polymerase chain reaction single-strand conformation polymorphism analysis(PCR-SSCP) in gastric carcinoma. Results： ① The positive rates of p16 protein expression were 96.25％ (77/80) in normal gastric mucosa, 92.00％ (45/50) in dysplastic gastric mucosa, and 47.54％ (58/122) in gastric carcinoma. The positive rate of p16 protein expression in gastric carcinoma was significantly lower than that in normal gastric mucosa and in dysplastic gastric mucosa (P<0.05). ② The positive rate of p16 protein expression in mucoid carcinoma (10.00％ ,1/10) was significantly lower than that of poorly differentiated carcinoma (51.22％ ,21/41), undifferentiated carcinoma (57.69％ ,15/26), and signet ring cell carcinoma (62.50％ ,10/16) (P< 0.05). ③ The positive rates of p16 protein in 30 cases paired primary and lymph node metastatic gastric carcinoma were 46.67 ％ (14/30) in primary gastric carcinoma,16.67％ (5/30) in lymph node metastatic gastric carcinoma. The positive rate of lymph node metastatic carcinoma was significantly lower than that of primary carcinoma(P<0.05). ④ Evaluation of mutation and deletion of p16 gene： There was no mutation of p16 gene in exon 2, but there were 5 cases displayed deletion of p16 gene in exon 2 in the 25 primary gastric carcinoma. Conclusions： ① The expression loss of p16 protein is related to carcinogenesis, histopathological subtypes,and lymph metastasis of gastric carcinoma. ② The mutation of p16 gene in exon 2 may not be involved in gastric carcinogenesis. But the deletion of p16 gene in exon 2 might be involved in gastric carcinogenesis.

Objective To study the effect of octreotide on patients with postoperative acute adhesive small bowel obstruction. Method In this study, 87 patients with postoperative acute adhesive small bowel obstruction were divided into 2 groups: experimental group (46 patients) and control group (41 patients). Patients in the control group were treated with routine therapy, including gastrointestinal decompression, intravenous infusion, antibiotic and enema. Patients in the experimental group were treated with routine therapy plus somatostatin analogue (octreotide) 0.1 mg. ih q8 h. for 72 hour. The alleviation of abdominal symptom and sign and the possibility of surgical intervention are observed and compared.Results Compared to the control group, the obstruction in the experimental group alleviated significantly,the abdominal pain relieved, the amount of draining decreased, and the passage of gas was earlier.Conclusions Based on the routine therapy, the use of octreotide significantly relieves the symptoms of obstruction and shortens the course of conservative therapy.

Objective To study the topoanatomy of anterior perineal plane and adjoining tissue structure in the preparation of ultra-low anterior resection of the rectum. Methods Dissection was performed on 16 male cadavers of semi-pelvis sectioned in the saggital plane. Eight indexes were measured and recorded. Results Anterior perineal plane was clearly found in all 16 cadavers. The median distance of rectum-urethra (R-U) was 14 mm (ranging 10 -17 mm). The contour of perineal body was trapezoid,which was narrow cranially and broad caudally. The median width of cranial perineal body was 8 mm (ranging 6 -9 mm), while the median width of caudal perineal body was 21 mm (ranging 18 -23 mm).The median numerus of thickness of perineal body (TPB), thickness of puborectalis (TPR), arrterior wall of rectum (aPR) -D, pPR-D and width of pelvic diaphragm (WPD) were 20. 5 mm ( ranging 17 - 23 mm),12 mm(ranging 10 -16 mm), 25 mm(ranging 21 -27 mm), 20 mm(ranging 16 -23 mm) and 8 mm (ranging 6 - 10 mm) respectively. Conclusions Anterior perineal plane clearly exists, through which about 20 mm more length of the distal rectum is available which could increase the sphincter-saving rate in cases of low rectal carcinoma.

Objective To evaluate the role of nicotinamide adenine dinucleotide phosphate hydrogen (NADPH) oxidase in bupivacaine-induced injury to neurons.Methods SH-SY5Y cells were seeded in 96-well culture plates at a density of 5× 104 cells/well and randomly divided into 3 groups (n=24each) using a random number table:control group (group C),bupivacaine group (group B),and apocynin (NADPH oxidase inhibitor) + bupivacaine group (group A+B).The cells were cultured in a serum-free medium in group C.The cells were cultured in a serum-free medium containing 1 mmol/L bupivacaine in group B.In group A + B,the cells were cultured for 30 min in a serum-free medium containing apocynin 100 μmol/L,and then cultured in a serum-free medium containing 1 mmol/L bupivacaine.At 2,4 and 6 h of incubation,the cells in 6 wells of each group were selected to evaluate the cell viability by MTS assay.At 4 h of incubation,the cells in 6 wells of each group were selected to detect reactive oxygen species (ROS) level by flow cytometry.Results Compared with group C,the cell viability was significantly decreased at 4 and 6 h of incubation,and the production of ROS was increased at 4 h of incubation in group B (P＜ 0.05).Compared with group B,the cell viability was significantly increased at 4 and 6 h of incubation,and the production of ROS was decreased at 4 h of incubation in group B (P＜0.05).Conclusion NADPH oxidase is involved in bupivacaine-induced injury to neurons.

Objective To explore the clinical effect of laparoscopic colorectal radical resection combined with simultaneous laparoscopic or open major hepatectomy for synchronous colorectal liver metastases (SCRLM).Methods The retrospective cohort study was adopted.The clinical data of 14 patients with SCRLM who were admitted to the Peking University Third Hospital from July 2010 to September 2015 were collected.Seven patients undergoing total laparoscopic colorectal radical resection combined with major hepatectomy (TLCRMH) were allocated into the TLCRMH group and 7 patients undergoing laparoscopic colorectal radical resection combined with open major hepatectomy (LCROMH) were allocated into the LCROMH group.The statuses of colorectal cancer and metastatic lesions were detected by endoscopy and imaging examination,and diagnostic and therapeutic plans were confirmed through the consultation of muhidisciplinary team.During the hepatectomy,total liver ultrasonography was performed and the extent of liver resection was above 3 hepatic segments.The follow-up of outpatient reexamination was applied to all the patients once every 3 months within postoperative year 2 and once every 6 months after 2 years till December 2015.(1) During operation,method of liver resection,radiofrequency ablation (RFA),operation time,volumes of intraoperative blood loss and blood transfusion,pathological results of primary lesions (T stage,N stage,nerve invasion and canalis haemalis invasion) were collected.(2) After operation,duration of intensive care unit (ICU) care,time for fluid diet intake,postoperative alanine transaminase (ALT),total bilirubin (TBil),complications and duration of hospital stay were collected.(3) Survival of patients and recurrence of tumor were followed up.Count data were analyzed by the chi-sqaure test.Measurement data with normal distribution were presented as x ± s and analyzed using the t test.Measurement data with skewed distribution were described as M(Qn) and M(range) and analyzed using the Mann-Whitney U test.The survival curve was drawn by the Kaplan-Meier method,and overall survival rate and tumor-free survival rate were calculated.The survival analysis was done using the Log-rank test.Results (1) The status of operation:1 and 6 patients in the TLCRMH group underwent respectively left and right hemihepatectomies and 7 patients in the LCROMH group underwent right hemihepatectomy.Four and 6 patients in the TLCRMH and LCROMH groups received local resection of liver metastatic lesions or RFA at segment Ⅱ,Ⅲ or Ⅳ of liver,respectively.The operation time,volumes of intraoperative blood loss and blood transfusion were (651 ± 218)minutes,(1 387 ± 871)mL,(914 ±641)mL in the TLCRMH group and (535 ± 83) minutes,(1 357 ±991)mL,(857 ± 360) mL in the LCROMH group,respectively,with no significant difference between the 2 groups (t =1.320,0.060,0.206,P ＞ 0.05).The numbers of patients in T2,T3,T4,N0,N1,N2 stages,with nerve invasion and canalis haemalis invasion were 1,5,1,3,4,0,3,2 in the TLCRMH group and 0,4,3,1,4,2,2,3 in the LCROMH group,respectively,showing no significant difference between the 2 groups (x2=2.111,3.000,0.311,0.311,P ＞ 0.05).(2) After operation,time for fluid diet intake,ALT,TBil,number of patients with complications and duration of hospital stay were (4.3 ± 1.0) days,(105 ± 47) U/L,(34 ± 25) μmol/L,3 (Ⅲ a,Ⅲ b and Ⅳ a grades of Dindo-Clavien grade),(27 ± 21)days in the TLCRMH group and (4.3 ± 1.1)days,(113 ± 57)U/L,(26 ± 11) μmol/L,4 (Ⅰ,Ⅰ,Ⅱ and Ⅳ a grades of Dindo-Clavien grade),(19 ± 9)days,respectively,showing no significant difference between the 2 groups (t =0.079,-0.286,0.806,X2 =0.286,t =0.856,P ＞ 0.05).The duration of ICU care in the TLCRMH and LCROMH groups were (2.1 ± 1.6) days and (1.0 ± 0.6) days,with a significant difference between the 2 groups (t =1.804,P ＜ 0.05).(3) The status of follow-up:all the patients were followed up for 3-54 months,and the median follow-up time was 15 months (range,3-39 months) in the TLCRMH group and 30 months (range,11-54 months) in the LCROMH group.The 1-,3-year overall survival rates were 100.0％ and 100.0％ in the TLCRMH group and 85.7％ and 64.3％ in the LCROMH group,respectively,showing no significant difference between the 2 groups (x2 =0.676,P ＞ 0.05).The postoperative 1-,2-year cumulative tumor-free survival rates and overage cumulative tumor-free survival time were 64.3％,64.3％ and 20.5 months in the TLCRMH group and 42.9％,14.3％ and 10.9 months in the LCROMH group,respectively,showing no significant difference between the 2 groups (x2=3.160,P ＞ 0.05).Conclusion TLCRMH is safe and feasible for patients with SCLM,and it is comparable with LCROMH in the incidence of postoperative complication and long-term outcomes.

Objective Streptococcus sanguis is a possible candidate bacterium for the caries replacement therapy, which has no advantages in the acidic environment.The aim of the study was to construct acid-resistant strains of Streptococcus sanguis, determine its acid tolerance, and explore the mechanism of its antagonism against Sterptococcus mutans.Methods By gradually reducing the pH value of the medium, we constructed acid-resistant strains of Streptococcus sanguis, observed their growth and measured their acid tolerance according to their survival rate against lethal pH.We evaluated the competitive relationship between Streptococcus sanguis and Streptococcus mutans by plate experiment and detected the changes of related acid resistance genes by real-time quantitative PCR.Results The growth of Streptococcus sanguis and its acid-resistant strains were limited by the pH value, and that of Streptococcus sanguis was better in either acidic or normal environment.The lethal pH value of Streptococcus sanguis was 3.6, that of its acid-resistant strains was 2.3, and the survival rate of the acid-resistant strains was 66.59% in the pH 3.6 environment.In comparison, the lethal pH value of Streptococcus mutans was 2.5, that of its acid-resistant strains was 2.1, and the survival rate of the acid-resistant strains was 2.55% in the pH 2.5 environment.In the presence of chloramphenicol, the acid-resistant strains could not survive in the original lethal pH.In the sub-lethal pH environment, the expressions of the acid resistance-related genes Groel and Dnak in the acid-resistant strains were significantly up-regulated as compared with those in the original Streptococcus sanguis (P<0.05).Conclusion Streptococcus sanguis has an acid adaptability and can enhance acid resistance in the sub-lethal pH environment.Acid-resistant Streptococcus sanguis in the replacement therapy may provide some new ideas for the treatment of dental caries.