Objective To evaluate the effect of hydrogen on the postoperative cognitive function in aged mice.Methods Sixty aged male Kunming mice,aged 12-14 months,weighing 35-45 g,were assigned into 3 groups (n=20 each) using a random number table:sham operation group (group Sham),partial hepatectomy group (group PH) and partial hepatectomy plus hydrogen-enriched saline group (group PH-H).On preoperative day 3,during operation and on postoperative day 3,hydrogen-enriched saline 10 ml/kg was intraperitoneally injected once a day in group PH-H,and the equal volume of normal saline was intraperitoneally injected once a day in S and PH groups.The spatial probe test was performed on postoperative day 7 to evaluate the cognitive function.Ten mice were selected on postoperative days 3 and 7 (after the end of the spatial probe test),and blood samples were collected for determination of serum tumor necrosis factor-alpha (TNF-α),interleukin-lbeta (IL-1β) and high-mobility group box 1 protein (HMGB1) concentrations by enzyme-linked immunosorbent assay.The animals were sacrificed after blood sampling,brains were removed and hippocampi were isolated for detection of the expression of TNF-α,IL-1β and HMGB1 by Western blot.Results Compared with group S,the percentage of time of staying at the target quadrant and the number of crossing the platform within 1 min were significantly decreased,the serum concentrations of TNF-α,IL-1β and HMGB1 were increased,and the expression of hippocampal TNF-α,IL-1β and HMGB1 was up-regulated in group PH (P＜0.05),and no significant change was found in the variables mentioned above in group PH-H (P＞0.05).Compared with group PH,the percentage of time of staying at the target quadrant and the number of crossing the platform within 1 min were significantly increased,the serum concentrations of TNF-α,IL-1β and HMGB1 were decreased,and the expression of hippocampal TNF-α,IL-1β and HMGB1 was down-regulated in group PH-H (P＜0.05).Conclusion Hydrogen can improve the postoperative cognitive function in aged inice.

Objective Autoimmune inner ear disease is one of the inner ear diseases that can be effectively treated clinically, and researches on it have significant clinical and practical value.This study explored the immunoregulating and therapeutic effects of the mi-croRNA-146a recombinant lentiviral vector (RLV) on immune-medicated inner ear disease (IMIED) in guinea pigs so as to avoid the adverse reactions of the currently used immunodepressants. Methods IMIED models were established in 30 guinea pigs by immuni-zation with keyhole limpet hemocyanin and divided into three groups: microRNA-146a RLV, empty lentiviral vector ( ELV) control, and surgery simulation control, microinjected via the scala tympani with microRNA-146a RLV, ELV, and PBS, respectively.Before and after immunization and at 7 days after microinjection, the auditory function and the level of specific anti-KLH antibodies in the ser-um were measured by auditory brainstem response ( ABR) audiometry and ELISA, respectively.Seven days after microinjection, 3 ani-mals in each group were subjected to HE staining and light microscopy, another 3 to fluoro-autography for realizing the situations of the inner ear transfected by the lentiviral vector, and the other 4 to measurement of the contents of microRNA-146a in the inner ear tissue. Results Compared with the baseline, immunization significantly increased the level of specific anti-KLH antibodies in the serum (0.09 ±0.01 vs 1.90 ±0.74 in the RLV group, 0.11 ±0.02 vs 2.20 ±0.75 in the ELV group, and 0.11 ±0.02 vs 2.10 ±0.64 in the surgery simulation control) as well as the average threshold of the ABRⅢwave (left ear 11.67 ±2.58 vs 61.67 ±5.16 and right ear 12.50 ±2.73 vs 60.00 ±4.47 in the RLV group;left ear 14.16 ±3.76 vs 64.33 ±9.17 and right ear 13.33 ±2.58 vs 60.83 ± 4.92 in the ELV group;left ear 15.83 ±3.76 vs 64.17 ±10.2 and right ear 15.00 ±5.47 vs 62.50 ±9.35 in the surgery simulation control) .The average threshold of the ABRⅢwave was decreased after local injection into the inner ear as compared with that after immunization, with no statistically significant difference between the ELV and surgery simulation control groups.Fluoro-autographic ob-servation showed that the main parts of the inner ear transfected with microRNA-146a recombinant lentiviruses were the spiral limbus, spiral ganglion afferent fibers, basal cells of the spiral ligament, Corti organ, and psalterial cords.The immune inflammatory reaction of the inner ear was significantly reduced in the RLV group as compared with the ELV and surgery simulation control groups, while the content of microRNA-146a in the inner ear tissue was obviously increased in the former group than in the latter two. Conclusion The microRNA-146a recombinant lentiviral vectors are widely distributed and transfected in the inner ear tissue after injected via the scala tympani.Injecting recombinant microRNA-146a lentiviral vectors into the inner ear can significantly reduce pathological damage and auditory dysfunction caused by inner ear immune inflammatory reaction.

Objective:To compare the wear resistance between dentin and dental restoration material in vitro.Methods:The friction and wear behaviors of natural tooth dentin respectively against highly polished polymer ceramic,glass ceramic and zirconia were investigated in an artificial saliva test environment by UMT-2 friction and wear testing machine.Worn surface morphology was observed by scanning electron microscopy(SEM),roughness was measured by roughness instrument,the rigidity value was weighed by electronic balance.Results:The roughness of the 3 materials was similar to that of dentin(P ＞ 0.05),the rigidity of dentin was less than that of the 3 materials (P ＜ 0.05).After friction and wear test,the abrasion quantity of dentin respectively against the 3 materials was less than that of the materials (P ＜ 0.05).There was positive correlation between the wear loss and the hardness of the 3 materials and dentin(r =0.846).The mass loss of the dentin against Poly Ceramic after grinding abrasion was the closest among the 3 materials.Conclusion:Different materials have varying degrees abrasion against dentine.The Polymer Ceramic has closer abrasion performancez with dentin than the other 2 materials.

Objective To study the distribution and clinicopathological characteristics between VEGF-C and pefitumoral lymph vessels density(PLVD) in breast cancer tissue, and to investigate the development and the mechanism of breast cancer-related lymphoedema (BCRL). Methods VEGF-C and VEGFR-3 were detected by using immunohistochemical technique for the detection of VEGF-C and its receptor VEGFR-3 in forty-seven breast cancer specimens. We measured the patients' circumferences of bilateral upper limbs to determine whether there was lymphoedema and made classification in the follow-ups. Results VEGF-C was positive in 33 out of 47 cases. PLVD significantly increased in VEGF-C positive groups (30.39±10. 46) than in negative groups (23.16±11.67) (P<0.05). VEGF-C semi-quantitative score was in a positive correlation with PLVD (r=0.334). The positive expression rate (42.55%) and semi-quantitative score (3.68±1.59) of VEGF-C increased in the lymph node positive group than in the negative group, PLVD increased in the lymph node positive group compared with that in negative group (32.12±10.29 vs. 24.82±11.06), P<0.05. The risk of lymphoedema increased in the VEGF-C negative group (5/14) compared with that in the positive group (3/33) (P<0.05). Conclusion VEGF-C has a high rate of positive expression in breast cancer, and is positively correlated with PLVD. High expression of VEGF-C can reduce the risk of BCRL in breast cancer.

Objective To examine the effect of modified electroconvulsive therapy (MECT) on the plasma oxida-tive stress level in bipolar depression. Methods Forty-two patients with bipolar depression were randomly divided into two groups. The intervention group (n=18) received antidepressants and 12 times MECT for 6 weeks and the control group (n=24) received antidepressants and Li2CO3 for 6 weeks. The Chinese versions of the 17 items Hamilton Depression Rating Scale (HAMD-17), Young Mania Rating Scale (YMRS), Clinical Global Impression Scale (CGI-S) and Treatment Emergent Symptom Scale (TESS) were used to assess participants at baseline and after 6 weeks of treatment. The plasma levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA)were detected at baseline and after 6 weeks to assess the level of oxidative stress. Results Repeated measures analysis of variance showed that the plasma level of SOD was higher in MECT group than in Li2CO3 group (F=15.26, P<0.01), and the level of MDA was higher in Li2CO3 group (F=18.18, P<0.01). The interactive effect of group and time was significant in GSH-Px level (F=6.39, P=0.02). The level of GSH-Px was lower in MECT group than in Li2CO3 group after 6 weeks (P<0.05). The CAT level was higher in the response patients than in non-response patients after 6 weeks (P<0.05). Con-clusions Both MECT treatment and Li2CO3 treatment can alter oxidative stress levels in patients with bipolar depression. The mechanisms underlying its therapeutic regimen may correct the imbalance of the plasma CAT level.

Objective To explore the current status and influencing factors of oral health knowledge and behavior among the elderly living in Gongshu District of Hangzhou City.Methods A total of 600 elderly individuals were selected by multi-stage stratified cluster random sampling and interviewed with a self-designed questionnaire which included demographic characteristics and oral health knowledge and behavior.The status and influencing factors of oral health behavior were analyzed by single and multiplefactor analysis methods.Results A questionnaire survey was conducted among 600 elderly residents,with a response rate of 99.3％ (596/600) and an effective response rate of 94.1％ (561/600).The rate of good oral hygiene was 50.4％.Those elderly with different age,education level,medical insurance and oral hygiene showed significant difference in qualified rate of oral health behavior (x2 values were 10.79,21.32,5.72 and 16.33,respectively; all P＜0.05).In Logistic regression model,education level was positively correlated with oral health behavior among the elderly,and the qualified rate of oral health behavior of the elderly with education level of junior school or above was 2.69 times higher than that of illiterate elderly (x2=10.53,P=0.001).Conclusion The awareness rate of oral health knowledge among the elderly living in Gongshu District of Hangzhou City is at a relatively higher level,though oral health behavior is moderate.Age,education level,medical insurance and oral hygiene could be impacting factors of oral health behavior.

Objective To observe the effects of MSH2 gene re expression on estrogen-induced apoptosis of colon cancer cells LOVO,and to explore its mechanisms.Methods According to different plasmid and whether with estradiol intervention,colon cancer LOVO cells were divided into empty plasmid with ethanol group,empty plasmid with estradiol group,MSH2 with ethanol group,MSH2 with estradiol group,estrogen receptor (ER) β with ethanol group,ERβ with estradiol group,ERβ with MSH2 and ethanol group and ERβ with MSH2 and estradiol group,and received corresponding treatment.The expression of MSH2,ERβ protein and apoptosis related caspase 3 protein were detected by Western blotting.Cell viability was measured by cell counting kit-8.Cell DNA fragments of each group were isolated with apoptosis DNA fragments isolation kit.And the DNA ladder was observed.The rate of apoptosis was detected by flow cytometer.Single factor variance analysis was performed for comparison among multiple groups,and t test was used for comparison between the two groups.Results After transfection,the expression of the MSH2 and ERβ at protein level in LOVO cells significantly increased and neither of their expression was effected by estradiol.The expression levels of caspase 3 cleavaged active fragments of ERβ with estradiol group and ERβ with MSH2 and ethanol group were higher than other groups,and there was no significant difference between these two groups.The LOVO cell viability of empty plasmid with ethanol group,empty plasmid with estradiol group,MSH2 with ethanol group,MSH2 with estradiol group,ERβ with ethanol group,ERβ with estradiol group,ERβ with MSH2 and ethanol group and ERβ with MSH2 and estradiol group was 1.72 ±0.25,1.74 ± 0.31,1.77 ± 0.35,1.74±0.33,1.70±0.34,1.02±0.48,1.71±0.31 and 1.07±0.18,respectively,and the differences between the groups were statistically significant (F=3.791,P＜0.05).Among them,the LOVO cell viability of ERβ with estradiol group was lower than that of ERβ with ethanol group,accordingly,that of ERβ with MSH2 and estradiol group was lower than that of ERβ with MSH2 and ethanol group,that of ERβ with estradiol group was lower than that of empty plasmid with estradiol group,that of ERβ with MSH2 and estradiol group was lower than that of MSH2 with estradiol group,and the differences were statistically significant (t=3.158,3.075,3.648,3.253,all P＜0.05).DNA ladder formed from DNA fragments of apoptosis cells was seen in ERβ with estradiol group and ERβ with MSH2 and estradiol group.The apoptosis rate of empty plasmid with ethanol group,empty plasmid with estradiol group,MSH2 with ethanol group,MSH2 with estradiol group,ERβ with ethanol group,ERβ with estradiol group,ERβ with MSH2 and ethanol group and ERβ with MSH2 and estradiol group was 7.86±0.19,7.87±0.39,8.39±1.02,9.05±1.54,7.54±0.99,19.77±2.35,7.76±1.32 and 19.30±1.75,respectively,and the differences between groups were statistically significant (F=45.436,P＜0.05).Among them,the apoptosis rate of ERβ with ethanol group was lower than that of ERβ with estradiol group,that of ERβ with MSH2 and ethanol group was lower than that of ERβ with MSH2 and estradiol group,that of empty plasmid with estradiol group was lower than that of ERβ with estradiol group,that of ERβ with MSH2 and estradiol group was lower than that of MSH2 with estradiol group,and the differences were statistically significant (t =8.260,9.133,8.596,7.617,all P＜ 0.05).Conclusions Estrogen may promote colon cancer cell apoptosis through ERβ pathway.The process of apoptosis maybe related with caspase protein,MSH2 may not be involved in the regulation of this signal pathway.

Refractoriness of acute myeloid leukemia (AML) cells to chemotherapeutics represents a major clinical barrier. Suicide gene therapy for cancer has been attractive but with limited clinical efficacy. In this study, we investigated the potential application of herpes simplex virus thymidine kinase/ganciclovir (HSV-TK/GCV) based system to inhibit chemoresistant AML cells. We first generated Ara-C resistant K562 cells and doxorubicin-resistant THP-1 cells. We found that the HSV-TK/GCV anticancer system suppressed drug resistant leukemic cells in culture. Chemoresistant AML cell lines displayed similar sensitivity to HSV-TK/GCV. Moreover, HSV-TK/GCV killing of leukemic cells was augmented to a mild but significant extent by all-trans retinoic acid (ATRA) with concomitant upregulation of Connexin 43, a major component of gap junctions. Interestingly, HSV-TK/GCV killing was enhanced by expression of vesicular stomatitis virus G glycoprotein (VSV-G), a fusogenic membrane protein, which also increased leukemic cell fusion. Co-culture resistant cells expressing HSV-TK and cells stably transduced with VSV-G showed that expression of VSV-G could promote the bystander killing effect of HSV-TK/GCV. Furthermore, combination of HSV-TK/GCV with VSV-G plus ATRA produced more pronounced antileukemia effect. These results suggest that the HSV-TK/GCV system in combination with fusogenic membrane proteins and/or ATRA could provide a strategy to mitigate the chemoresistance of AML.
Cell Fusion ; Cell Line ; Coculture Techniques ; Connexin 43 ; Cytarabine ; Gap Junctions ; Genetic Therapy ; Glycoproteins* ; Homicide* ; K562 Cells ; Leukemia, Myeloid, Acute ; Membrane Proteins ; Simplexvirus* ; Suicide ; Thymidine* ; Tretinoin ; Up-Regulation ; Vesicular Stomatitis*

Bacterial biofilms have been demonstrated to be closely related to clinical infections and contribute to drug resistance. Berberine, which is the main component of Coptis chinensis, has been reported to have efficient antibacterial activity. This study aimed to investigate the potential effect of a combination of berberine with ciprofloxacin (CIP) to inhibit Salmonella biofilm formation and its effect on expressions of related genes (rpoE, luxS, and ompR). The fractional inhibitory concentration (FIC) index of the combination of berberine with CIP is 0.75 showing a synergistic antibacterial effect. The biofilm's adhesion rate and growth curve showed that the multi-resistant Salmonella strain had the potential to form a biofilm relative to that of strain CVCC528, and the antibiofilm effects were in a dose-dependent manner. Biofilm microstructures were rarely observed at 1/2 × MIC/FIC concentrations (MIC, minimal inhibition concentration), and the combination had a stronger antibiofilm effect than each of the antimicrobial agents used alone at 1/4 × FIC concentration. LuxS, rpoE, and ompR mRNA expressions were significantly repressed (p < 0.01) at 1/2 × MIC/FIC concentrations, and the berberine and CIP combination repressed mRNA expressions more strongly at the 1/4 × FIC concentration. The results indicate that the combination of berberine and CIP has a synergistic effect and is effective in inhibiting Salmonella biofilm formation via repression of luxS, rpoE, and ompR mRNA expressions.
Anti-Infective Agents ; Berberine ; Biofilms ; Ciprofloxacin ; Coptis ; Drug Combinations ; Drug Resistance ; Drug Resistance, Multiple ; Repression, Psychology ; RNA, Messenger ; Salmonella

Objective To compare the difference of cutpoint and clinical significance of HbA1C for the diagnosis of abnormal glucose metabolism in two population groups with different ages.Methods According to oral glucose tolerance test(OGTT),the cutpoint and clinical significance of HbA1C for the diagnosis of type 2 diabetes and impaired glucose regulation(IGR)were investigated in the two population groups.Results The mean HbA1C of 1 064 young subjects in an academy and 1 671 aged subjects in a community were 5.31% ±0.41% and 5.79% ±0.71%,respectively.The cutpoints of HbA1C for diagnosis of diabetes were 5.7%(specificity 86.7%,sensitivity 66.7%)and 5.9%(specificity 73.8%,sensitivity 80.1%)in the two population groups,and 5.6% for diagnosis of IGR (specificity 82.8%,sensitivity 55.8%)and 5.7%(specificity 60.9%,sensitivity 64.3%),respectively.87.8%,78.7%,and 38.5% were diagnosed diabetes by current OGTT criteria at HbA1C levels of ≥5.7%,≥5.9%,and≥6.5%,IGR being 61.6%,39.6%,and 4.1%,and normal glucose tolerance being 24.4%,10.0%,and 0.4%.Conclusion The cutpoints of HbA1C for diagnosis of diabetes and IGR are different in populations with different ages and HbA1C levels.As one of diagnostic criteria for diabetes,HbA1C 6.5% with relatively higher specificity and lower sensitivity must be combined with fasting blood glucose,random blood glucose,and OGTT.