OBJECTIVETo explore the expression of Ghrelin and high mobility group protein B1 (HMGB1) in the serum and the intestinal tissue of sepsis model rats, and to evaluate the effect of electro-acupuncture (EA) at Zusanli (ST36) on the expression of HMGB1 and Ghrelin.

METHODSForty-eight male Wistar rats were randomly divided into four groups, i.e., the sham-operation (sham), the cecal ligation and puncture group (CLP), the CLP + EA at Zusanli (ST36) group (EA), and the CLP + Ghrelin receptor blocking agent + EA group (GHSRA), 12 in each group. A sepsis rat model was prepared by CLP. The incision of the abdominal wall was immediately sutured along the ventral midline for rats in the Sham group. In the EA group EA at Zusanli (ST36) was performed 20 min after CLP surgery with the constant voltage (2 - 100 Hz, 2 mA) for 30 min. In the GHSRA group, Ghrelin receptor blocking agent, [D-Arg1, D-Phe5, D-Trp79, Leu11]-substance P (700 nmol/kg), was administered through intravenous injection immediately after CLP, and 20 min later, EA at Zusanli (ST36) was performed in the same way as for rats in the EA group. Blood samples were withdrawn 12 h after CLP. The serum levels of Ghrelin and HMGB1 were detected using ELISA. Ghrelin expressions and the number of Ghrelin immunopositive cell in the jejunum were determined by immunohistochemistry. HMGB1 contents of the jejunum tissue were detected by Western blotting.

RESULTSCompared with the Sham group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly increased and levels of Ghrelin and the expression rate of immunopositive cells significantly decreased in the CLP group (P < 0.05). Compared with the CLP group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly decreased, but levels of Ghrelin and the expression rate of immunopositive cells significantly increased in the EA group (P < 0.05). Compared with the EA group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly increased in the GHSRA group (P < 0.05), but there was no statistical difference in levels of Ghrelin between the two groups (P > 0.05). The serum level of HMGB1 was negatively correlated with Ghrelin in the Sham group, the CLP group, and the EA group (r = -0. 528, P < 0.01).

CONCLUSIONSEA at Zusanli (ST36) could inhibit the expression of HMGB1 in the jejunum of septic rats, and promote the expression of Ghrelin. The expression of HMGB1 was inhibited by Ghrelin receptor blocking agent, which suggested that the anti-inflammation of EA at Zusanli (ST36) might be associated with Ghrelin.

Animals ; Disease Models, Animal ; Electroacupuncture ; Ghrelin ; metabolism ; HMGB1 Protein ; metabolism ; Jejunum ; metabolism ; Male ; Rats ; Rats, Wistar ; Sepsis ; metabolism

Objective To study the influence of electro-acupuncture(EA)at Zusanli points(足三里穴) on the apoptosis of thymocytes in rats with abdominal infection and its mechanism.Methods A total of 40 Sprague-Dawley(SD)rats were randomly divided into four groups,including normal control group,model group,non-acupoint group and Zusanli group.The abdominal infection model of rat was made by cecal ligation and puncture(CLP).After abdominal cavity infection for 36 hours,the apoptosis of thymocytes was observed under electron microscope and light microscope,and the apoptosis ratio of thymocytes was determined by Annexin V-PI method with flow cytometry technique.The content of Bcl-2 protein of thymocytes and concentration of corticosterone in plasma were determined.Results Abdominal infection resulted from CLP could significantly increase the apoptosis of thymocytes and lead to the typical histopathological changes of apoptosis of thymocytes under electron microscope and light microscope.Apoptosis ratios of thyrnocytes in model group[(44.7?3.3)%],non-acupoint group[(42.7?3.0)%]and Zusanli group[(32.6?3.3)%] were significantly higher than the ratio in the control group[(21.2?2.3)%,all P0.05).Abdominal infection resulted from CLP also could reduce the content of Bcl-2 protein of thymocytes.The content of Bcl-2 protein of thymocytes in model group(71.2?5.6),non-acupoint group(73.5?5.9)and Zusanli group(82.4?6.8) were significantly lower than normal control group(95.3?6.3,all P

Objective To examine the expression of recombinant cytolethal distending toxin(CDT)produced by Actinobacillus actinomycetemcomitans(Aa).Methods CDT encoding gene cdtABC was amplified by PCR.Through TA clone and restriction endonuclease digestion,gene cdtABC and vector pQE60 were ligated to form pQE60-cdtABC expression system which transformed into competent cells.Protein expression was induced by IPTG and examined by SDS-PAGE and Western-blotting.Results Random colony PCR of pQE60-cdtABC transformed cells demonstrated that all strains contained cdtABC gene.The DNA sequence was blast with cdtABC gene from GenBank and 99%homology was obtained.SDS-PAGE and Western-blotting confirmed that recombinant CDT was obtained.Conclusions CDT protein expression system was reconstructed and recombinant protein was obtained.

OBJECTIVETo study the mechanism and significance of peripheral blood mononuclear cell (PBMC) of neonates infected with hepatitis B virus (HBV).

METHODSEighty-four HBsAg-positive and HBeAg-negative mothers and their newborns were recruited in this study. Sixteen hepatitis B virus markers (HBVM)-negative mothers and their neonates were served as control. All these cases had no symptoms of hepatitis, serious pregnancy complications and preexisting disease. Age, gestational age and the method of delivery were matched in two groups (P > 0.05). Five ml blood samples were taken from the peripheral vein of the pregnant women before delivery and from neonates within 24 hours after birth, before inoculation of HBV vaccine (HBVac). Serum and PBMC were isolated from 2 ml and 3 ml samples respectively. The sera, PBMC and the last supernatant of PBMC washing were stored at -80 degrees C. HBVM of neonates were detected by using enzyme linked immunosorbent assay (ELISA). HBV DNA in serum, PBMC and the last supernatant of PBMC washing of mothers and neonates were detected by using a nested-polymerase chain reaction (n-PCR). Two pairs of oligonucleotide primers, the outer primer pair for first PCR and inner primer pair for second PCR, designed according to region S of HBV genome were synthesized at Shanghai Cell Biology Institute of Chinese Academy of Sciences. The neonates who were HBV DNA positive in PBMC but HBsAg and HBV DNA negative in serum were followed up for one year, HBsAb in serum and HBV DNA in PBMC were observed in the neonates.

RESULTS(1) The positive rate of HBV DNA in 84 serum and PBMC of mothers were 53.57% and 40.48%, respectively (chi(2) = 2.891, P > 0.05). All the results were weakly positive. (2) Twenty-four (28.57%) newborns in the study group were infected, including 7 who were only HBV DNA positive in serum, 11 only HBV DNA positive in PBMC and 6 in both, all the results were weakly positive. HBsAg was negative in all the newborns. None of the neonates in control group was infected with HBV. There was significant difference between the two groups (chi(2) = 4.55, P < 0.05). (3) Of all the study cases, 11 (13.10%) neonates were HBV DNA weakly positive in PBMC but HBsAg and HBV DNA negative in serum. Of their mothers, 5 were only HBV DNA positive in serum, 2 only positive in PBMC and 4 positive in both serum and PBMC. Seven of the 11 neonates were followed up for one year and at the end of follow-up, 4 were HBsAb positive and HBV DNA negative in PBMC; 3 were HBsAb negative, and among the 3 cases HBV DNA in 2 was still positive in PBMC, HBsAg and HBV DNA in serum were negative in all the 7 neonates.

CONCLUSION(1) HBV DNA positivity either in serum or in PBMC in mothers can result in infection of PBMC with HBV in their neonates. (2) PBMC infection with HBV can exist for a long time in neonates while HBsAg and HBV DNA are negative in serum, and may result in vaccination failure in neonates.

Case-Control Studies ; DNA, Viral ; blood ; Female ; Hepatitis B ; diagnosis ; immunology ; Hepatitis B Vaccines ; administration & dosage ; Hepatitis B virus ; Humans ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; Leukocytes, Mononuclear ; virology ; Pregnancy

Objective:To explore the value of different echocardiographic indices on the detection of left ventricular ejection fraction (LVEF) in patients with left ventricular anterior and anterolateral wall motion abnormalities .Methods:Twenty-one patients with left ventricular anterior and anterolateral wall motion abnormalities were enrolled from Jan .2010 to Dec .2015 .The left ventricular ejection fraction (LVEF) of all subjects were measured by M-mode echocardiography ,two-dimensional echocardiography and apical biplane Simpson’s method ,and were compared with left ventricular angiography . Results:There were obvious differences in LVEF evaluation between left ventricular angiography method and M-mode echocardiography or two-dimensional echocardiography (P<0 .05) ,while the correlation between the angiographic method and apical biplane Simpson’s measurement was good .Further the subgroup analysis with the M-mode measurement of 55% as the boundary showed that there were no significant differences among the four kinds of methods when LVEF is more than or equal to 55% .And in patients with LVEF less than or equal to 55% ,LVEF measured by two-dimensional method and M-mode was higher than that of left ventricular angiography (P< 0 .05) ,and there was no statistical difference between Simpson's method and left ventricular angiography (P=0 .061 1) .Conclusions:For patients with left ventricular anterior and anterolateral wall motion abnormalities ,M-mode or two-dimensional echocardiography can be directly adopted when LVEF is more than or equal to 55% measured by M-mode;but if LVEF is less than or equal to 55% ,apical biplane Simpson’s method is recommended for determination .

OBJECTIVETo investigate the correlation between moderately and severely chronic periodontitis and coronary heart disease, as well as the role of fibrinogen in the mechanisms responsible for the correlation between periodontitis and coronary heart disease.

METHODS95 subjects who were systemic health or patients of coronary heart disease with or without periodontitis were enrolled. All the subjects were placed into 4 groups based on their periodontal status and cardiovascular health. The 4 groups were healthy control group (HC), moderately and severely chronic periodontitis group (MSP), coronary heart disease group(CHD), and MSP coexisted with CHD group (MSP+CHD). Clinical periodontal index were examined, at the same time, plasma fibrinogen levels and serological changes used in diagnosing of cardiovascular disease routinely were determined. ANOVA and ANCOVA were used in the statistical analysis.

RESULTSFibrinogen levels of HC, MSP, CHD, and MSP+CHD group were (2.36+/-0.37), (3.63+/-0.73), (4.08+/-0.84), and (4.14+/-0.96) g/L, respectively. Fibrinogen levels of MSP and MSP+CHD group were significantly higher than that of healthy controls (P<0.01). The patients with moderately to severely chronic periodontitis were more likely to have coronary heart disease as compared to periodontally healthy controls (OR=2.527, P=0.047) after adjusted for blood pressure and body mass index.

CONCLUSIONModerately and severely chronic periodontitis maybe a risk factor of coronary heart disease and fibrinogen could be one of the biological basis which links periodontitis with coronary heart disease.

Chronic Periodontitis ; Coronary Disease ; Female ; Humans ; Male ; Middle Aged ; Periodontal Index ; Periodontitis ; Risk Factors

OBJECTIVEPorphyromonas gingivalis (P. gingivalis) is considered to be major putative periodontal pathogens. The purpose of the study was to evaluate P. gingivalis and clinical effects of scaling and root planning (SRP) in 20 subjects after 3 months.

METHODSTwenty periodontitis patients were selected. The mean age was (44.33 +/- 13.86) years old. Clinical assessments of probing depth (PD), clinical attachment loss (CAL) and bleeding on probing (BOP) were made prior to SRP and at 3 months post-therapy. Subgingival plaque samples were taken at each visit and analyzed using TaqMan real-time polymerase chain reaction for the presence and levels of P. gingivalis. The quantification for P. gingivalis was also performed with the help of the species-specific primers/probes and the serial dilution of the plasmid standards.

RESULTSMean probing depth, mean clinical attachment loss and bleeding on probing showed significant reduction at 3 months (P<0.001). The prevalence and level of P. gingivalis were significantly reduced after SRP (P<0.001). A positive correlation was found between the numbers of P. gingivalis and PD at baseline (P<0.001). There were no correlation between the initial level of P. gingivalis at baseline and the clinical improvement after therapy. But the number of P. gingivalis at responding sites was more decreased than non-responding sites after SRP (P<0.05).

CONCLUSIONSRP produced a good clinical improvement. The prevalence and level of P. gingivalis were significantly reduced after SRP. The effect of SRP may be determined by the degree of P. gingivalis is decreased. The real-time polymerase chain reaction can be used to evaluate the effect of periodontal therapy.

Adult ; Dental Plaque ; Dental Plaque Index ; Dental Scaling ; Female ; Humans ; Male ; Middle Aged ; Periodontal Attachment Loss ; Periodontal Pocket ; Periodontitis ; Porphyromonas gingivalis ; Root Planing

OBJECTIVETo investigate the relationship between plasma levels of fibrinogen, the-beta455 G/A fibrinogen gene polymorphism and the severity of periodontal inflammation and to explore the possible role of fibrinogen in the association of periodontitis with coronary heart disease (CHD).

METHODSA total of 121 patients with moderate to severe periodontitis and periodontally healthy and gingivitis controls were enrolled in the study. Peripheral blood samples were collected and the plasma fibrinogen levels were determined by the clotting method of Clauss. Polymerase chain reaction and restriction fragment length polymorphism analysis with Hae III were used to examine the -beta455 G/A fibrinogen gene polymorphism.

RESULTSFibrinogen levels were significantly higher in moderately or severely chronic periodontitis patients [(3.45 +/- 0.68) g/L] than periodontally healthy and gingivitis controls [(2.47 +/- 0.42) g/L, P < 0.001]. The carrier status of the A allele at position -455 in the beta fibrinogen gene was associated with elevated fibrinogen levels and the frequency of the-A455 allele in the beta fibrinogen gene in the patient group was significantly higher than in the control group (P = 0.032). Carriers of the -A455 allele were about 3-fold more likely to have moderate or severe periodontitis as compare to individuals without the -A455 allele( OR = 3. =135, P= 0.008).

CONCLUSIONSFg-beta455 G/A polymorphism may contribute to the elevated plasma fibrinogen levels and put individuals at higher risk of having severe periodontitis. As the independent risk factor of CHD, fibrinogen levels and Fg-beta455 G/A polymorphism may play a role in the pathogenesis of periodontitis.

Adult ; Alleles ; Case-Control Studies ; Chronic Periodontitis ; genetics ; Coronary Disease ; genetics ; Female ; Fibrinogen ; analysis ; genetics ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length

OBJECTIVEThe aim of this study was to investigate the prevalence of T. forsythensis in Chinese chronic periodontitis patients, and correlate it with clinical parameters.

METHODS108 chronic periodontitis patients were selected according to clinical criteria. In each patient, subgingival plaque samples were collected from two of the periodontitis involved molar sites with the deepest periodontal pocket depth(diseased sites) and one healthy gingival sulcus (healthy sites). T. forsythensis was detected by 16S rRNA polymerase chain reaction. Clinical assessments including probing depth, clinical attachment loss and bleeding on probing, were made at sampling sites.

RESULTSThe prevalence of Tforsythensis in diseased sites and healthy sites was 59.72% and 3.70% respectively (P < 0.001). The prevalence of T. forsythensis was significantly higher in deeper pockets and more serious clinical attachment loss, Spearman rank correlation coefficient was 0.48 and 0.51 respectively. The prevalence of T. forsythensis in BOP (+)sites was 61.31%, and in BOP(-) sites was 8.80% (P < 0.001).

CONCLUSIONThe prevalence of T. forsythensis was significantly correlated with clinical parameters, suggesting that T. forsythensis is closely related to chronic periodontitis in the Chinese population.

Adult ; Bacteria ; Chronic Periodontitis ; Dental Plaque ; Female ; Gingiva ; Humans ; Male ; Middle Aged ; Periodontal Pocket ; Periodontitis ; Polymerase Chain Reaction ; Prevalence ; RNA, Ribosomal, 16S

OBJECTIVETo investigate the prevalence of H. actinomycetemcomitans in Chinese chronic periodontitis (CP) patients and periodontally healthy adults.

METHODS116 chronic periodontitis patients and 111 periodontally healthy adults were included. In each CP patient, subgingival plaque samples were collected from two sites of different molars with the greatest probing depth (PD) and one periodontally healthy site (PD < or =3 mm). The samples of periodontally healthy adults were obtained from the mesio-buccal site of one first upper molar. Bacteria DNA were extracted for detection of H. actinomycetemcomitans by 16S rRNA PCR.

RESULTSThe prevalence for H. actinomycetemcomitans of diseased sites (33.62%) was significantly higher than that of healthy sites from CP patients (0.86%) and the periodontally sites (0.90%) (P < 0.01). No significant difference was observed between male and female CP patients (P > 0.05). A decreasing trend of H. actinomycetemcomitans was observed as the age increased. And the pocket depth and clinical attachment losswas associated with the occurrence of H. actinomycetemcomitans in a positive mode. And H. actinomycetemcomitans was more often detected in the bleeding sites on probing.

CONCLUSIONH. actinomycetemcomitans was more frequently detected in periodontitis sites than periodontally healthy sites. For CP patients, a higher prevalence was associated with the seriously involved sites than those moderate and mild implicated sites. H. actinomycetemcomitans is considered to be the one of the periopathogens involved in the etiology of chronic periodontitis.

Adult ; Aggregatibacter actinomycetemcomitans ; Chronic Periodontitis ; DNA, Bacterial ; Dental Plaque ; Female ; Healthy Volunteers ; Humans ; Male ; Periodontitis ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S