Objective:To explore the effects ofbufalin (BUF) combined with doxorubicin (DOX) on the proliferation and apoptosis in human lung cancer cell line A549 in vitro.Methods:Methyl thiazolyl tetrazolium (MTT) assay was used to measure the inhibitory effects of BUF,DOX and their combination on the growth ofA549 cells.Hoechst 33342 staining was used to observe the changes of nucleus.Flow cytometry was used to investigate the apoptosis and cell cycle distribution of A549 cells.Western blot was used to examine the expression of apoptotic protein.Results:BUF and DOX showed inhibitory effect on the A549 cells in a dose and time-dependent manner.Compared with BUF or DOX alone,combination of BUF (1,20,100 nmol/L) with DOX (1.0 μg/mL) could significantly increase the growth inhibition rate ofA549 cells at 24,36,72 h,respectively (all P＜0.05).BUF and DOX alone could induce apoptosis,and their combination could significantly increase the apoptosis ratio.In addition,BUF combined with DOX could block the cell stage of A549 cells,keep the cell stage stay in S stage and up-regulate the expression of caspase-3.Conclusion:BUF combined with DOX can significantly inhibit the proliferation ofA549 cells,which might be related to the induction of apoptosis,cell cycle S phase arrest and caspase-3 up-regulation.

Objective: To observe the rheological properties of octoxynol-9 vaginal thermosensitive in situ gel [(O-9)-VTG] for predicting the gelation behavior in vivo, and evaluate the retention ability in vagina.Methods: The rheological parameters of (O-9)-VTG and (O-9)-VTG diluted by stimulant vaginal fluid (SVF) were measured by a Haake Rheomix to characterize the rheological properties.The vaginal samples after the administration of self-made (O-9)-VTG and O-9 gel were withdrawn, and then the concentration of octoxynol-9 in the samples was determined to evaluate the retention ability.Results: (O-9)-VTG was Newtonian fluid with low viscoelasticity under room temperature and converted to gel at 32.6℃.The formula could still transform into gel at body temperature after diluted by SVF, and resided in the vagina of mice above 8 h.Conclusion: (O-9)-VTG has suitable gelation temperature and rheological properties.Compared with the self-made octoxynol-9 gel, (O-9)-VTG has satisfactory retention in vagina, which meets the requirements for vaginal topical use.

Objective:To prepare compound mupirocin film forming gel and establish the quality control methods. Methods:The compound film forming gel was prepared using tannic acid and salicylic acid as the esterifying agents interacting with hydroxypropyl cel-lulose followed by cross-linking with tartaric acid and sorbitol to form a new film forming material, and then antimicrobial mupirocin and anesthetic dyclonine were added to prepare compound mupirocin film forming gel. The contents of mupirocin and dyclonine were deter-mined by HPLC. The column was Hypersil ODS2 (250 mm × 4. 6 mm, 5 μm) and the mobile phase consisted of methanol-ammonium acetate buffer (1. 15 g ammonium acetate, 300 ml water, 1 ml glacial acetic acid) (75 :25). The flow rate was 1. 0 ml·min-1 and the detection wavelength was 230 nm. The column temperature was 30℃ and the injection volume was 20μl. Results:The film form-ing gel was yellow, brown and gelatinous at the normal temperature, and formed transparent film after coating on the skin. The linear range of mupirocin and dyclonine was 20-400μg﹒ml-1(r=0. 9999) and 10-200μg﹒ml-1(r=0. 9996), respectively. The aver-age recovery was 99.9%(RSD=0.54%, n=9) and 99.6%(RSD =1.45%, n =9), respectively. Conclusion: The preparation process is reasonable, simple and controllable in the preparation of ideal film forming gel.

Objective To analyze the risk factors of mortality in hospitalized patients with peptic ulcer bleeding (PUB).Methods From January 2003 to December 2012,1 210 patients with PUB were collected.Among them,1 170 patients were cured (cured group) and 40 patients died (dead group).The general information and clinical data of patients were collected,which included gender,age,smoking history,non-steroidal anti-inflammatory drugs intake,haematemesis,shock,blood infusion and rebleeding.The information of patients combined with other diseases was also collected.The lab findings and gastroendoscopy findings were also collected,including hemoglobin,platelets,serum albumin,blood urea nitrogen,serum creatinine levels,coagulation,location of ulcer and Forrest classification.Chi-square test was performed for comparison between groups of count data.The t-test was used for comparison between groups of measurement data and normally distributed,and Mann-Whitney rank sum test was used for non-normal distribution.The risk factors of mortality of patients with PUB were analyzed by univariate and multivariate Logistic regression analysis.Results The rates of age over 65,combined with other diseases,shock,rebleeding,abnormal coagulation,Forrest classification above Ⅱ b,medicine spraying to stop bleeding under gastroendoscope,gastroendoscopic hemostasis with titanium clip and operation of dead group (57.5％,23/40; 27.5％,11/40; 25.0％,10/40; 42.5％,17/40; 25.0％,10/40; 35.0％,14/40; 15.0％,6/40; 12.5％,5/40 and 17.5％,7/40) were all higher than those of cured group (25.0％,293/1 170; 7.4％,86/1 170; 12.5％,146/1 170; 13.1％,153/1 170; 5.1％,60/1 170; 20.9％,244/1 170; 4.8％,56/1 170; 4.1％,48/1 170 and 6.5％,76/1 170).The differences were statistically significant (x2 =21.117,18.651,5.400,27.728,9.203,4.613,6.332,4.661 and 5.710,all P＜0.05).The serum albumin level of dead group ((28.71±7.13) g/L) was lower than that of cured group ((32.82±7.55) g/L) and the difference was statistically significant (t=2.215,P＜0.05).Between the groups,there were no significant differences in gender distribution,rate of patients smoking,rate of patients taking non-steroidal anti-inflammatory drugs,rate of patients with haematemesis,volume of blood infusion,location of ulcer,dosage of proton pump inhibitor (PPI),average hemoglobin level,blood urea nitrogen level,serum creatinine and platelet count (all P＞0.01).Age over 65,combined with other diseases and rebleeding were the independent risk factors of mortality of patients with PUB during hospitalization (OR=4.821,12.959 and9.627,all P＜0.01).Conclusion Age over 65,combined with other diseases and rebleeding are the independent risk factors of mortality of patients with PUB during hospitalization.

Objective To evaluate the role of cyclooxygenase-2 (COX-2) in ventilator-induced lung injury in rats.Methods Thirty healthy adult male Sprague-Dawley rats,aged 3.0-3.5 months,weighing 300-350 g,were randomly divided into 3 groups (n =10 each) using a random number table:traditional tidal volume group (group T,VT =8 ml/kg),large tidal volume group (group L,VT =40 ml/kg) and NS398 (selective COX-2 inhibitor,VT =40 ml/kg) group (group N).In group N,8 mg/kg NS398 (in 2 ml of 10％ dimethyl sulfoxide) was injected intraperitoneally at 1 h before ventilation,while dimethyl sulfoxide 2 ml was administrated instead of NS398 in T and L groups.After 4 h of mechanical ventilation,arterial blood samples were obtained for blood gas analysis and PaO2 was recorded.The animals were sacrificed and lungs removed for microscopic examination and for determination of wet/dry lung weight ratio (W/D ratio) and concentrations of total protein,intercellular adhesion molecule-1 (ICAM-1),tumor necrosis factor-alpha (TNF-α),NO and 6-keto-prostaglandin F1alpha (6-keto-PGF1 α) in bronchoaveolar lavage fluid (BALF).Pulmonary permeability index (PPI) and TXB2/6-keto-PGF1 α ratio were calculated.Results Compared with group T,PaO2 was significantly decreased,W/D ratio and PPI were increased,the total protein,ICAM-1,TNF-α and NO levels and TXB2/6-keto-PGF1 α ratio in BALF were increased in group L(P ＜ 0.05).PaO2 was significantly higher,W/D ratio and PPI were lower,and total protein,ICAM-1,TNF-α and NO levels and TXB2/6-keto-PGF1 α ratio in BALF were lower in group NS than in group L (P ＜ 0.05).The damage to lung tissues was severe in group L,and obviously alleviated in group N.Conclusion COX-2 is involved in ventilator-induced lung injury in rats.

Objective To evaluate the efficacy of 10-day compound allantoin containing quadruple regimen in the treatment of chronic gastritis with Helicobacter pylori (H.pylori) infection,and to compare with the bismuth-containing quadruple therapy.Methods Altogether 173 patients with H.pylori positive chronic gastritis confirmed by gastric endoscope were divided into 10-day compound allantoin containing quadruple regimen group (n =43),24-day compound allantoin containing quadruple regimen group (n =46),10-day bismuth-containing quadruple regimen group (n =42) and 24-day bismuth-containing quadruple regimen group (n =42).After the treatment,the eradication rate of H.pylori,the rate of gastrointestinal symptoms (epigastric pain,bloating and belching) relief and the adverse effects of each group were observed.Intention-to-treat (ITT),per-protocol (PP) statistical analysis and chi-square analysis were performed for statistical analysis.Results H.pylori eradication rates of 10-day compound allantoin containing quadruple regimen group,24-day compound allantoin containing quadruple regimen group,10-day bismuth-containing quadruple regimen group and 24-day bismuth-containing quadruple regimen group analyzed by ITT were 90.7％ (39/43),91.3％ (42/46),90.5％ (38/42) and 88.1％ (37/42),respectively; while analyzed by PP were 90.7％ (39/43),93.3％ (42/45),90.5％ (38/42) and 90.2％ (37/41),respectively.And there were no statistical differences between groups (all P＞0.05).Ten days after the treatment,the rates of epigastric pain relief of 24-day compound allantoin containing quadruple regimen group and 24-day bismuth-containing quadruple regimen group were 81.1％ (30/37) and 78.8％ (26/33),respectively,the rates of bloating relief were 82.4％ (28/34) and 71.0％ (22/31),respectively,and the rates of belching relief were 76.9％ (20/26) and 75.0％ (21/28),respectively.There were no statistical differences between the two groups (all P＞ 0.05).However after 24-day treatment,the rates of epigastric pain relief of 24-day compound allantoin containing quadruple regimen group and 24-day bismuth-containing quadruple regimen group were 91.9 ％ (34/37) and 87.9％ (29/33),respectively,the rates of bloating relief were 94.1％ (32/34) and 87.1％ (27/31),respectively,and the rates of belching relief were 96.2％ (25/26) and 85.7％ (24/28),respectively.There were no statistical differences between the two groups (all P＞0.05).And the rates of epigastric pain and bloating relief increased after 24-day treatment compared with those of 10-day treatment,however the differences were not statistically significant between the two groups (all P＞ 0.05).In 24-day compound allantoin containing quadruple regimen group,the rate of belching relief was higher after 24-day treatment compared with that of 10-day treatment,and the difference was statistically significant (x2=4.127,P=0.042).No severe adverse effects were observed in each group,and there were no adverse effects such as oral metal odor,tongue black and melena in compound allantoin containing quadruple therapy.Conclusions Ten-day compound allantoin containing quadruple therapy as first-line approach in the treatment of chronic gastritis with H.pylori infection can get better H.pylori eradication and the efficacy is similar to bismuth quadruple therapy.Meanwhile the symptom relief rate is high and no obvious adverse effects were found.

As a novel concept for cell delivery,cell sheet may retain the extracellular matrix and adhesive proteins,avoid the use of bioma-terials for delivery,and increase cell survival rate while reduce cell loss following cell transplantation.This review summarizes the use of cell sheet technology for periodontal and pulp-dentin complex regeneration,highlights recent progresses and future challenges in this field.

Objective To evaluate the role of spinal phosphatidyl-inositol 3-kinase/Akt (PI3k/Akt) signaling pathway in the maintenance of bone cancer pain (BCP) in rats and its relationship with microglial activation.Methods Forty healthy female Sprague-Dawley rats,weighing 180-200 g,were randomly divided into 5 groups (n =8 each):sham operation group (group S) ; PI3K inhibitor LY294002 group (group L) ; group BCP; BCP + dimethyl sulfoxide (DMSO) group (group BCP + D) ; BCP + LY294002 group (group BCP + L).BCP was induced by inoculating Walker 256 mammary gland carcinoma cells into the medullary cavity of the left tibia.At 7-9 days after inoculation,LY294002 2.5 μg/10 μl was injected intrathecally in L and BCP + L groups,normal saline 10 μl was injected intrathecally in S and BCP groups,and 5％ DMSO 10 μl was injected intrathecally in BCP+ D group once a day.Mechanical paw withdrawal threshold (MWT) was measured at 1 day before inoculation and 1,3,5,7,8 and 9 days after inoculation.The rats were sacrificed after MWT was measured on day 9 after inoculation and the L4-6 segments of the spinal cord were removed to determinate the activation of spinal microglia using immunofluorescence.Results Compared with group S,MWT was significantly decreased,and the activation of spinal microglia was increased in BCP,BCP + D and BCP+ L groups.Compared with BCP and BCP + D groups,MWT was significantly increased,and the activation of spinal microglia was decreased in BCP + D group.Conclusion Spinal PI3K/Akt signaling pathway is involved in the maintenance of BCP possibly through activating microglia in spinal dorsal horns of rats.

Along with recent advances in biological signal molecule and tissue engineering technology,periodontal regeneration has been gained more and more new opportunities,but also faces many challenges.This paper briefly reviewes the preclinical and clinical studies of periodontal tissue regeneration,highlighting the latest achievement and progress in the clinical study of biological signal molecules and stem cell therapy in the treatment of periodontal disease worldwide.

Objective To quantify the changes of tongue color with the development of the disease according to the characteristics of human vision.Methods Psychological scales of tongue color perception were obtained from thc psychophysical experiments.Polynomial regression and support vector regression (SVR) were used to establish the mathematical model between the physical values and the psychological scales of tongue color.Results The psychological scales exported from the model could correspond to the visual perception of tongue color change, and SVR has higher accuracy.Conclusions The psychological scale, can not only quantitatively evaluate the tongue color in the development of the disease, but also quantify the degree of disease, to assist the doctors for disease diagnosis and treatment.