Objective To investigate the effect of hydrogen peroxide (H2O2) pretreatment on free mitochondrial cyto-chrome c (Cyt-c) release in mitochondrial levels, and reveal the mechanism of the ischemic preconditioning (IPC) on the ischemia reperfusion (IR) injury thereof. Methods The rat liver mitochondria was isolated and made free mitochondria. Free mitochondria were divided into 5 groups:control group (C) and different concentrations of Ca2+groups (12.5, 25, 50 and 100 μmol/L). The levels of Cyt-c and second mitochondria-defived activator of caspase (Smac) were detected after 10 min stimulation of free mitochondria. The free mitochondrial IPC reperfusion model was made and divided into seven groups:C group, IR group and different concentrations of H2O2 groups (2 μL H2O2 in 200 μL system respectively, final concentration of 1, 2, 5, 20 and 100 μmol/L respectively). 100 μmol/L Ca2+was used again on the simulation of IR group. The level of Cyt-c release was detected. The changes in the activity of cardiolipin were detected in IR group and H2O2 (1 and 2 μmol/L of final concentration) groups.Results Compared with C group, there were significantly higher levels of Cyt-c and Smac emission in 25, 50, and 100 μmol/L Ca2+groups (P<0.05). Compared with IR group, there was significantly decreased level of Cyt-c emission in H2O2 (1 and 2 μmol/L) groups (P < 0.05). The activity of cardiolipin was changed when reducing release of Cyt-c. Conclusion Cyt-c was bonded with cardiolipin more closely when the low concentration of H2O2 pretreatment in mitochondria. There was a lower level of Cyt-c emission in mitochondria after stimulation with high concentration of Ca 2+(100 μmol/L Ca2+). The blocking apoptotic pathway plays a key fact in the effect of IPC on IR injury.

Objective To study the effect of periodontitis on rats with chronic bacterial prostatitis. Methods A total of 80 male rats were randomly divided into the 4 weeks group (n=40) and the 8 weeks group (n=40), and then the two groups were randomly divided into the normal control group (N=10), the periodontitis group (PE=10), the chronic bacterial prostatitis group (CBP=10), and the peri-odontitis+chronic bacterial prostatitis group (CBP+PE=10) respectively. The pathological changes, inflammation score, level of TNF-αand IL-1β, and indicators of periodontal of all rats were observed. Results In the 4 weeks group, the indicators of periodontal in PE group and CBP+PE group were higher than that in N group and CBP group (P<0. 05), but there was no significant difference between PE group and CBP+PE group as well as between the N group and the CBP group (P>0. 05). The pathological changes, inflammation score,TNF-αlevel ,IL-1 β level in CBP+PE group and CBP group were higher than that in N group and PE group (P<0. 05), while there was no sing-nificant difference between N group and PE group as well as between CBP+PE group and CBP group (P>0. 05). In the 8 weeks group, the indicators of periodontal in PE group and CBP+PE group were higher than that in N group and CBP group (P<0. 05), but there was no sig-nificant difference between PE group and CBP+PE group as well as between the N group and the CBP group (P>0. 05). The pathological changes, inflammation score, TNF-α level , IL-1 β level in CBP +PE group and CBP group were higher than that in N group and PE group (P<0. 05),while there was no singnificant difference between N group and PE group (P>0. 05). The pathological changes, inflam-mation score,TNF-α level ,IL-1 β level in CBP+PE group were higher than those in the CBP group (P<0. 05). Compared between 4 weeks group and 8 weeks group, there was no obvious difference in N group in terms of periodontal indexes. Periodontal indexes in 8 weeks PE group was higher than that in 4 weeks PE group (P<0. 05), but there was no significant difference prostate tissue pathology, inflamma-tion score,TNF-α level,IL-1βlevel (P>0. 05). Pathology, inflammation score,TNF-αlevel, IL-1βlevel in 8 weeks CBP group were low-er compared to 4 weeks CBP group (P<0. 05), and there was no significant difference between the indexes of periodontal (P>0. 05). Prostate tissue pathology, inflammation score, TNF-αlevel, IL-1βlevel in 8 weeks CBP+PE group were lower than that in 4 weeks CBP+PE group (P<0. 05), but indicators of periodontal in 8 weeks CBP+PE group were higher than 4 weeks CBP+PE group (P<0. 05). Conclusion Chronic bacterial prostatitis combined with periodontitis can inhibit self-healing tendency of chronic bacterial prostatitis of rats and keep rats in chronic inflammatory phase.

Objective To investigate whether human breast milk may bind to hepatitis B surface antigen (HBsAg) and its characteristics.Methods Breast milk samples from five women with negative HBsAg and hepatitis B surface antibody (anti-HBs) at one to two months post delivery were fractioned into cream and skimmed milk by centrifugation.The human breast milk and each fraction as well as cow and goat milk samples,served as controls,were separately incubated with highly purified yeast recombinant HBsAg,followed by determination of their binding capability to HBsAg by enzyme linked immunosorbent assay (ELISA) and the inhibition rate for binding of HBsAg to anti-HBs by quantitative chemiluminescence microparticle immunoassay.After boiled for 1 min or pasteurized in 65 ℃ for 30 min,the thermal stability of the active components of milk was detected.One-way ANOVA and SNK tests were performed for statistical analysis.Results The operative concentration of HBsAg was 0.1 μg/ml.Breast milk from all five women showed significantly better binding capability to HBsAg than cow or goat milk (1.306±0.300 vs 2.157±0.150 and 2.232±0.093,F=34.303,P＜0.01).The quantitative experiments showed that the inhibition rate of human breast milk was higher than that of the control group [(74.26± 17.26)％ vs (0.00±5.50)％,F=57.806,P＜0.01].The binding ability to HBsAg of skimmed milk was comparable with that of whole milk,indicating milk protein(s) played critical roles in binding to HBsAg (0.877 ± 0.486 vs 0.513 ± 0.069 and 0.376 ± 0.146,F=44.475,P＜0.01).After boiled for 1 min or Pasteurization,the binding ability to HBsAg of whole breast milk remained,but that of skimmed milk went down (F=16.598,P＜0.01).Both whole breast milk and skimmed milk could inhibit the binding of HBsAg to anti-HBs (F=278.341 and 269.408,both P＜0.01).Conclusions The inhibition of binding to HBsAg by human breast milk indicates that human milk may interact with HBsAg.The active components mainly exist in milk proteins and are thermal stable.

OBJECTIVETo investigate whether programmed death ligand 1 (PD-L1) expressed in the periodontal tissue of chronic periodontitis and the correlativity of PD-L1 and different degrees of chronic periodontitis, provide experience for immunoregulation mechanism, clinical treatment and prognosis of chronic periodontitis.

METHODSGingiva and periodontal tissue of healthy people and chronic periodontitis patients were collected. Based on clinical probing, periodontal tissue were classified into three groups: periodontal tissues of healthy people, periodontal tissue of mild chronic periodontitis, periodontal tissue of severe chronic periodontitis. Fluorescent quantitation polymerase chain reaction was applied to explore the expression of PD-L1 mRNA in the periodontal tissue of the different groups. Western blot and immunohistochemistry method were utilized to test the expression of PD-L1 protein in the periodontal tissue of the different groups. Combining with clinical image data, the relationship between differentially expressions of PD-L1 and different degrees of chronic periodontitis was analyzed.

RESULTSThe relative expression quantity of PD-L1 in the periodontal tissue of the mild chronic periodontitis was significantly higher that of the severe chronic periodontitis (P<0.01). The relative expression quantity of PD-L1 in the periodontal tissue of healthy subjects and severe chronic periodontitis had no statistical significance (P>0.05).

CONCLUSIONThe expression of PD-L1 in the periodontal tissue negativelv regulates inflammatory periodontal tissue damage.

B7-H1 Antigen ; Chronic Periodontitis ; Gingiva ; Humans ; Polymerase Chain Reaction ; RNA, Messenger

Objective:To investigate the effect of blocking the expression of receptor activity modifying protein 1 (RAMP1 )on calcito-nin gene-related peptide(CGRP)-induced MG-63 cell proliferation.Methods:RAMP1 siRNA was synthesized and screened by tran-scription in vitro.The subcultured MG-63 cells were divided into the following groups:RAMP1 siRNA interference group,empty vector group and blank control group.The mRNA expression and the membrane distribution changes of the calcitonin receptor-like receptor (CRLR)and the receptor component protein (RCP)in MG-63 cells were examined by real-time PCR and immunofluorescence method respectively.Results:RAMP1 and CRLR mRNA and the fluorescence intensity of MG-63 cells decreased after transfection by RAMP1 siRNA(P <0.05).In RAMP1 interference group,the expression of RCP mRNA and the fluorescence intensity were higher than those in the other two groups(P <0.05).After RAMP1 siRNA interference,the proliferation of MG-63 cells was inhibited(P <0.05). Conclusion:RAMP1 siRNA transfection may reduce CRLR expression and inhibite the proliferation of MG-63 cell.

OBJECTIVETo evaluate the inhibitory effect of high mobility group chromosomal protein N2 (HMGN2) on human tongue carcinoma tumor in nude mice.

METHODSA transplantation tumor model in nude mice was constructed by injecting Tca8113 cells. After a week, negative control groups, masculine control groups, and HMGN2 groups were established. Cell culture of the three groups were separately injected with washing buffer II, cis-dichlorodiamineplatinum (DDP), and HMGN2 protein. The tumors were moved after four treatments, and then analyzed by hematoxylin-eosin (HE) staining.

RESULTSA transplanted tumor model was established successfully. The volumes of HMGN2 groups and masculine control groups were smaller than those of the negative groups. Mouse weight did not differ among the three groups. Average tumor weight of the negative group was (0.38 +/- 0.19)g, that of the HMGN2 group was (0.21 +/- 0.15)g, and that of the DDP group was (0.23 +/- 0.16)g. These factors indicated no statistically significant difference among the three groups. The tumor inhibitory rate of HMGN2 group was 45.71%, and that of the positive group was 39.44%. Based on evaluation by naked eye, the tumor in the negative group was larger than that in other groups. In addition, cell necrosis was observed during HE staining.

CONCLUSIONHMGN2 could significantly inhibit growth of the transplanted tumor in nude mice.

Animals ; Carcinoma ; Cell Line, Tumor ; High Mobility Group Proteins ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Tongue Neoplasms

Objective To optimize the prescription of GA and GB hydrophilic gel matrix tablets; To study the in vitro release mechanism. Methods On the basis of the results of the mono-factor investigation, mixture uniform design was used to optimize the handicraft molding prescription of GA and GB hydrophilic gel matrix tablets. The release mechanism was investigated by the vitro of the GA and GB hydrophilic gelmatrix tablets to accumulate releasing rate to conduct linear fitting. Results The optimized prescription of GA and GB hydrophilic gel matrix tablets was: powder: HPMC: lactose=23:24:53. Conclusion Mixture uniform design can be used to optimize the prescriptions of GA and GB hydrophilic gel matrix tablets, and the results are accurate. The hydrophilic gelmatrix tablets release medicine by non-Fick mechanism, and the medicine release is in accordance with zero-order.

Objective To analyse clinical application of hepatitis B virus core antibody(HBcAb)detected by using the chemilu-minescence microparticle immunoassay.Methods A total of 1 6 830 specimen with positive HBcAb detected by using the two pairs of semi-hepatitis test from January 2012 to November 2014 were collected,and divided into three groups according to the cut off in-dex(COI)of detection results of HBcAb,including group 1.0-<9.0,group 9.0-<1 1.0 and group COI≥1 1.0,and detection re-sults were statistically analysed.The hepatitis B virus(HBV)DNA test was carried out in specimen with negative hepatitis B surface antigen(HBsAg)and hepatitis B surface antibody (HBsAb)and COI≥1 1.0.Results The detection rate of HBsAg(+)HBsAb(-) (13.84%)was significantly higher than other expression patterns in group ≥1 1.0(P <0.05).There was no statistically significant differences in positive rate among all expression patterns of HBsAg and HBsAb in the group 9.0-<1 1.0(P >0.05).The detec-tion rate of HBsAg(+)HBsAb(-)of group 9.0-<1 1.0 was significantly lower than that of the other two groups(P <0.05).A total of 304 specimen were HBsAg(-)HBsAb(-)and COI≥1 1,among them 64 specimen were HBV DNA postive and the posi-tive rate was 21.0%.Conclusion In the detection of HBcAb,COI≥1 1 and 1.0-<9.0 could be reference indicators for diagnosiing current and past HBV infection respectively,which should be combined with other laboratory indicators of HBV clinical data for comprehensive analysis.

Objective To observe the effects of low frequency ultrasound on carotid artery plaque and artery stenosis. Methods 156 patients with carotid atherosclerosis were divided into treatment group (n=80) and control group (n=76). The control group was administered routine medicine, while the treatment group accepted low frequency ultrasound therapy in addition. The size and shape of carotid artery plaque, severity of stenosis and the level of lipid were observed before and after treatment, and the side-effects were recorded. Results The intima-media thickness (IMT), diameter of plaque, plaque score decreased after treatment in both groups, and decreased more in the treatment group than in the control group (P<0.05); while the frequence of moderate stenosis and severe stenosis was less (P<0.05). The levels of low density lipoprotein- cholesterol and total cholesterol decreased in both groups after treatment (P<0.05), and decreased more in the treatment group than in the control group (P<0.05). No serious side-effect was observed. Conclusion Low frequency ultrasound can reduce the atherosclerotic plaques in carotid artery and relieve the stenosis.

Objective To evaluate the long-term effect of surgical procedures for congenital choledochal cyst (CCC).Methods From 1986 to 2000, 120 cases of CCC were admitted and 73 of them underwent the primary operations in General Hospital of PLA. Three types procedures were performed,type I: external drainage of CCC in 7 cases; type II:cystojejunal Roux-en-Y anastomosis in 5 cases; type III: cyst excision with cystojejunal Roux-en-Y anastomosis or cystoduodenostomy in 57cases,and other procedures in 4 cases.Results 68 cases were followed-up for 6 months to 5 years (median 2.7 years). Three cases undergoing type I operations accepted reoperations;two cases undergoing type II operations accepted reoperations due to severe complications as cholongitis and hepatolithiasis; 57 cases treated by type III operation with the good results 88.7% and none reoperation.Conclusions External drainage is only a first-aid management on emergency basis. Internal drainage should never be done,because the effect is temporary,and severe complications result in reoperations. Cyst excision with biliary tract reconstruction is recommended as the optimal treatment of CCC.