OBJECTIVETo compare the protein profiles between decapeptide-treated and untreated planktonic cells of Streptococcus mutans (S. mutans) by differential proteomic analysis to determine and identify the key proteins.

METHODSIn our previous study, we investigated decapeptide (KKVVFKVKFK-NH2), which was a novel adenosine monophosphate. Compared with other oral pathogens tested, decapeptide had a preferential antibacterial activity against S. mutans. It also inhibited S. mutans biofilm formation and reduced the one-day developed biofilm. In the present study, we first synthesized decapeptide, and then compared the protein profiles between decapeptide-treated and untreated planktonic cells of S. mutans by two-dimensional gel electrophoresis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. We also verified different expressions of key protein enolase in the protein level.

RESULTSThe results showed that decapeptide altered the protein expression of planktonic S. mutans. These proteins were functionally involved in carbohydrate degradation by glycolysis, protein folding, conjunction, transport, translation, adenosine triphosphate binding, protein binding, sequence-specific DNA binding, transcription factor activity, and two-component response regulator activity. Western blot results showed that enolase protein expression decreased obviously in decapeptide-treated cells of S. mutans.

CONCLUSIONThe protein expression of S. mutans significantly changed after synthetic antimicrobial decapeptide treatment, suggesting that decapeptide may present a preferential effect on oral caries by changing the expression of certain key proteins, such as enolase protein.

Anti-Bacterial Agents ; Anti-Infective Agents ; Biofilms ; Dental Caries ; Depsipeptides ; genetics ; Electrophoresis, Gel, Two-Dimensional ; Oligopeptides ; genetics ; Proteomics ; Streptococcus mutans ; metabolism

Objective:To optimize the alcohol precipitation technology of Qingmai granules. Methods:The relative density of liq-uor,the concentration and time of alcohol precipitation were chosen as the factors,the yield of dry extract and content of diosgenin as the indices,the alcohol precipitation technology was optimized by orthogonal test. Results:The optimal alcohol precipitation technology was as follows:the extraction liquid was concentrated till the relative density was 1. 13-1. 18 g·ml-1 ,ethanol was added until the concen-tration was up to 60% with the alcohol precipitation time of 24h. Conclusion:The optimized technology is stable,reasonable and feasi-ble,which can provide experimental basis for the clinical application of Qingmai granules.

Objective To study the expression and clinical significance of BRCA1 and BRCA2 in sporadic breast cancer in women of Jiaodong peninsula. Methods Immunohistochemistry and tissue array were used to detect the expression of BRCA1 and BRCA2 in 100 cases of sporadic breast cancer and 30 cases of benign breast tumor in women of Jiaodong peninsula. Results ① The expression rate of BRCA1 and BRCA2 was 49% (49/100) and 50% (50/100) in breast cancer, 80% (24/30) and 83.33% (25/30) in benign breast lesions respectively. The expression rate of BRCA1 and BRCA2 in breast cancer was lower than that in benign breast lesions (P<0.05). ② The expression of BRCA1 and BRCA2 was uncorrelated with factors such as tumor size, lymphatic metastases, age and menopause or not(P>0.05). ③ There was no dependency between the expression of BRCA1 and BRCA2 (P> 0. 05). Conclusions The expression rate of BRCA1 and BRCA2 in breast carcinoma in women of Jiaodong peninsula was lower than that in benign breast lesions, suggesting that the expression of BRCA1 and BRCA2 was related to the occurrence of sporadic breast carcinoma in women of Jiaodong peninsula. However, the role of BRCA1 and BRCA2 in the genesis and development of the breast carcinoma is independent.

OBJECTIVE:To establish the method for the content determination of Ciclosporin ophthalmic emulsion and to probe into its stability.METHODS:HPLC method was used to determine the content of ciclosporin.The separation was performed on Eclipse XDB-C8 column.The mobile phase consisted of acetonitrile-methanol-water(67.5 :5 :27.5) with the flow rate of 0.8 mL?min-1.The detection wavelength was set at 210 nm and column temperature was 55 ℃.Its stability was studied by classic isothermal accelerated test and highlight struck test.RESULTS:The linear range of ciclosporin was 5～60 mg?L-1(r=0.999 9) with average recovery of 100.39%(RSD=1.15%).The preparation was stable under heat and light,and its shelf life at 25 ℃ was predicted about 6.88 years.CONCLUSIONS:The method is accurate and feasible for the content determination of Ciclosporin ophthalmic emulsion.The preparation can be stored at room temperature.

OBJECTIVE:To observe the protective effect of the traditional Chinese medicine Yiqimingmu oral solution on photochemical damage of retina in rats. METHODS:SD rats were divided into four groups randomly:negative control group, model group, low dose Yiqimingmu oral solution group and high dose Yiqimingmu oral solution group. All the groups except the control one were continually exposed to Lux (1 900?106.9) green fluorescent light for 24 hours to establish retinal photochemical damage model. The low dose and high dose groups were administered intragastrically with Yiqimingmu oral solution (15mL?kg-1?d-1 and 30mL?kg-1?d-1) 7 days before light expose. Normal saline solution was administered intragastrically in control and model group. At 6 hours and 6, 14 days after light expose, the content of malandialdehyde (MDA) and activity of SOD in rats' retina were measured. The histopathologic changes were observed by transmission electron microscopy and light microscopy. RESULTS:At 6 and 14 days after light expose, the activity of SOD in Yiqimingmu solution high dose group were higher than in the model control group(P

Aim To study the distributive character of mitomycin(MMC) magnetic nanoparticles and MMC normal saline solution in mice.Methods A HPLC method for the determination of MMC in tissues and serum were established and applied to determine MMC in biological samples.Results Under the guidance of an external magnetic field,targeting rates of MMC to liver viscus are 82.72% after 30 minutes tail vein administration of(1 mg?kg~(-1)) mitomycin magnetic nanoparticles,it was 2.37 times larger than targeting rates of MMC normal saline solution group.The distributions of heart and kidney of mitomycin magnetic nanoparticles were less than that of MMC normal saline solution group.Compared with the results of tail vein administration mitomycin nanoparticles,interaction between the external magnetic field and magnetic nanoparticles is significantly effective to increase targeting rates of MMC to liver.Compared with the results of tail vein administration mitomycin magnetic nanoparticles without the external magnetic field,Mitomycin magnetic nanoparticles under the guidance of an external magnetic field is significantly effective to increase targeting rates of MMC to liver.Conclusions Mitomycin magnetic nanoparticles under the guidance of an external magnetic field is significantly effective to increase targeting rates of MMC to liver and prolonging effect on the action in vivo.

Objective The Rex shunt has been used to treat children with extrahepatic portal hypertension by creating a venous bypass between the superior mesenteric vein and the left portal vein to circumvent the extrahepatic portal venous obstruction.This retrospective study aimed to analyse our results in the use of this novel approach.Methods 52 patients with cavernous transformation and obstructed main portal veins were treated by Rex shunts.Results The age of children was 1.4 ～ 12 year,the mean age was 3.7 years.The patients were followed up from 1.5 to 5 years.In 48 patients,there was no recurrence of gastrointestinal bleeding after surgery making an efficacy rate of 92.3％.In the recurrence group,the postoperative venous pressure in the superior mesenteric vein was (27.6 ± 3.2) cmH2O (1 cmH2O =0.098kPa) which was significantly higher than the non-recurrence group (23.5 ± 3.1)cmH2O.The difference between the pre-and post-Rex shunting was significantly lower in the recurrence group (5.7 ± 1.8)cmH2O than the non-recurrence group (11.7 ± 3.3) cmH2 O,P ＜ 0.05.Thus,a low reduction in postoperative pressure was an early manifestation of poor prognosis.Conclusions The Rex shunt was safe and efficacious.The degree of reduction in postoperative venous pressure in the superior mesenteric vein could be used to predict recurrence of gastrointestinal bleeding.

Objective To establish a new rat model of biliary atresia by pure ethanol injection into the common bile duct.Methods A catheter was inserted and fixed in the common bile duct in male SD rats .Saline (8 rats) or pure ethanol (16 rats) was injected through the catheter ,respectively, and the biochemical and pathological changes in the rats were examined .Results SD rats in the experimental group were divided into a persistent injury and a restoration of liver dysfunction groups according to pathological and biochemical detection .In the persistent injury group , biochemical impair-ments were significantly higher at 8 weeks after ethanol injection than those in the control group and restoration group .Dis-tinct pathological changes in the liver were observed using HE , SMA, and Masson staining .Conclusions It is a reliable animal model of biliary atresia induced by injection of pure ethanol into the common bile duct in the rat .It will provide a useful tool in future studies of biliary atresia .

Objective To explore the clinical effect of periarticular knee fractures combined with anterolateral ligament (ALL) avulsion fracture.Methods From June 2014 to November 2015,13 patients with ALL avulsion fracture in periarticular knee fractures were fixed by the star plate,cannulated or screws suture anchor,including 9 male and 3 female with the mean age of 39.5 years (range from 28 to 51 years).6 patients were diagnosed distal femoral fracture,4 tibial plateau fracture,and 3 tibial intercondylar eminence avulsion fracture which all combined with ALL avulsion fracture.Postoperatively,all patients were treated with the adjustable knee brace for 3 months.Results All patients were followed up for 4-20 months,the mean follow-up time was 13.5 months.The average surgical time was 130 min (range from 90 to 210 min).In all 13 patients,ALL was found and identified.Bone union was obtained in all patients,and the bone union time was 11.4 weeks (10-12 weeks).7 cases were fixed with star plate,5 cases were fixed with hollow screw,and 1 case was fixed with anchor.All follow-up patients underwent anteroposterior and lateral X-rays.Pain,swelling and functional recovery of the knee joint were observed.Patients were assessed postoperatively with International Knee Documentation Committee (IKDC) score,Lysholm score,and Tegner activity scale to evaluate the clinical effect.At the last follow-up,all patients had full range of motion of the knee joint which from 0° to approximate 120°.The Lysholm,objective IKDC were 86.5 and 84.6 respectively.Among the 13 patients evaluated,9 patients were graded A,and 4 patients were graded B by subjective IKDC.The Tegner activity scale at the last follow-up was 6.7.Incision of poor healing in 1 case,and got healed after 32 d cleaning dressing.Conclusion This study confirmed the presence of the ALL.The fixation of ALL avulsion fracture in periarticular knee fractures can be an effective procedure at a minimum follow-up of 13.5 months.

Objective To study the differentiation and proliferation ability of the spinal neural stem cells (NSCs) at different gestational ages in fetal rats. Methods Sprague-Dawley fetal rats were divided into group A (12 days of pregnancy), group B (14 days of pregnancy) and group C (16 days of pregnancy). NSCs were separated with enzyme-assisted microdissection. The diameter and numbers of NSCs balls were measured at different time. The cell growth curve was drawn with CCK8 colorimeter. NSCs were identified with BrdU/Nestin immunohistochemical staining. They were induced with 10% fetal bovine serum for 10 days, and the expression of β-tubulinⅢ and glial fibrillary acidic protein was detected with immunocytochemistry. Results There were cells expressed BrdU, Nestin, β-tubulinⅢ and GFAP in all the group. The most cells (22.74±0.79%) expressed β-tubulinⅢ in the group B, but no significant difference between group B and group C. The cell vitality on the 5th day of third-generation neural stem cells was the most in group B. Conclusion For enzyme-assisted microdissection, it may obtain more neurons to isolate the neural stem cells from 14 days of pregnancy pregnant rats.