Association of Salivary Helicobacter pylori Infection with Oral Diseases: a Cross-sectional Study in a Chinese Population.

OBJECTIVE: The aim of this study was to detect the prevalence of oral H.pylori among adults and to investigate the correlation between H.pylori infection and common oral diseases. STUDY DESIGN: A cross-sectional study was performed among adults Chinese who took their annual oral healthy examination at The First Affiliated Hospital, Zhejiang University School of Medicine, China. RESULTS: The study included 1050 subjects in total and oral H.pylori infection occurred in 60.29% of the subjects. The prevalence rates of oral H.pylori in patients with periodontal diseases (63.42%) and caries (66.91%) were significantly increased than those without oral diseases (54.07%), respectively (P < 0.05), while the difference between subjects with recurrent aphthous stomatitis and controls was not significant. In addition, the differences of positive rates of H.pylori with or without history of gastric ulcer were statistically significant (69.47% vs 58.26%, P<0.05). Presenting with periodontal diseases (OR 1.473;95% CI 1.021 to 2.124), caries (OR 1.717; 1.127 to 2.618), and having history of gastric ulcer (OR 1.631; 1.164 to 2.285) increased the risk of H.pylori infection. CONCLUSIONS: Oral H.pylori infection is common in adult Chinese, which is significantly associated with oral diseases including periodontal diseases and caries.

Medium-chain fatty acid nanoliposomes suppress body fat accumulation in mice.

Medium-chain fatty acids (MCFA) are widely used in diets for patients with obesity. To develop a delivery system for suppressing dietary fat accumulation into adipose tissue, MCFA were encapsulated in nanoliposomes (NL), which can overcome the drawbacks of MCFA and keep their properties unchanged. In the present study, crude liposomes were first produced by the thin-layer dispersion method, and then dynamic high-pressure microfluidisation (DHPM) and DHPM combined with freeze-thawing methods were used to prepare MCFA NL (NL-1 and NL-2, respectively). NL-1 exhibited smaller average size (77.6 (SD 4.3) nm), higher zeta potential (- 40.8 (SD 1.7) mV) and entrapment efficiency (73.3 (SD 16.1) %) and better stability, while NL-2 showed narrower distribution (polydispersion index 0.193 (SD 0.016)). The body fat reduction property of NL-1 and NL-2 were evaluated by short-term (2 weeks) and long-term (6 weeks) experiments of mice. In contrast to the MCFA group, the NL groups had overcome the poor palatability of MCFA because the normal diet of mice was maintained. The body fat and total cholesterol (TCH) of NL-1 (1.54 (SD 0.30) g, P = 0.039 and 2.33 (SD 0.44) mmol/l, P = 0.021, respectively) and NL-2 (1.58 (SD 0.69) g, P = 0.041 and 2.29 (SD 0.38) mmol/l, P = 0.015, respectively) significantly decreased when compared with the control group (2.11 (SD 0.82) g and 2.99 (SD 0.48) mmol/l, respectively). The TAG concentration of the NL-1 group (0.55 (SD 0.14) mmol/l) was remarkably lower (P = 0.045) than the control group (0.94 (SD 0.37) mmol/l). No significant difference in weight and fat gain, TCH and TAG was detected between the MCFA NL and MCFA groups. Therefore, MCFA NL could be potential nutritional candidates for obesity to suppress body fat accumulation.

[Electroacupuncture-induced plasticity between hand and face representations in the motor cortex in healthy subjects].

OBJECTIVE: To observe the plasticity between hand and face representations of the motor cortex of healthy volunteers after electroacupuncture(EA) at Hegu(LI4), so as to provide a scientific basis for the theory of "Hegu is indicated for orofacial problems". METHODS: Using a cross-over design (self-controlled study), 10 healthy volunteers were randomly assigned to an acupuncture group and a sham acupuncture group (two-weeks wash-out period between the two groups). Subjects in the acupuncture group received EA stimulation (2 Hz, 0.5 to 1 mA, 30 min) at LI4 on their left hands. Adhesive pads sticked on the left hands of subject in the sham acupuncture group, thus, the placebo needle provided participants with a similar appearance to that in the acupuncture group but no skin penetration, and the placebo needles were connected to an EA device with a broken wire inside. Using transcranial magnetic stimulation technology, the motor evoked potentials (MEPs) of the first dorsal interosseous muscle and the orbicularis oculi muscle in hand and face representations in the contralateral motor cortex were recorded before and after EA and sham EA, and the total amplitude, effective stimulation area and center of gravity of MEPs were calculated. RESULTS: Compared with that before intervention, for acupuncture group, the total amplitude of MEPs in hand representation in the contrala-teral motor cortex was significantly increased(P<0.05),while the total amplitude of MEPs in face representation was significantly decreased (P<0.05). The effective stimulation area in hand representation was significantly increased(P<0.01), and there was no difference in face representation(P>0.05). The difference in the center of gravity of the X-axis in hand representation was statistically significant (P<0.05)，with the center of gravity moved an average of 0.6 cm to the outside, and there was no difference in face representation (P>0.05). There was no difference in the center of gravity of the Y-axis in hand and face representations(P>0.05). For sham acupuncture group, there were no differences in total amplitude of MEPs, effective stimulation area and the center of gravity in hand and face representations (P>0.05). CONCLUSION: EA at LI4 can induce plasticity between the hand and face representations of the motor cortex in healthy volunteers (exciting the hand representation of the motor cortex, while inhibiting the motor cortex representation), which provides a scientific basis for treating facial and mouth diseases by acupuncture at LI4 and the theory of selecting acupoints of the corresponding meridian distal to the disease location.

[Tissue engineered bone reconstruction with modified PLGA/Type-I collagen compound scaffold].

OBJECTIVE: To fabricate bone grafts by bone marrow stromal cell combined with modified PLGA/Type-I collagen compound scaffold using tissue engineering method. METHODS: The modified PLGA/Type-I collagen compound scaffold was fabricated. The rabbit primary cultured osteoblasts were identified and seeded onto the modified compound scaffold for one week in vitro. The adhesion and growth of cells were observed with scanning electron microscope. The complex of cells and scaffold was implanted into the subcutaneous region of rabbits and new bone formation was evaluated. RESULTS: The rabbit bone marrow stromal cells were induced and differentiated into osteoblasts. The adhesion and growth of osteoblasts in cluster were observed on the surface of scaffolds. New bone formation was observed at one month postoperatively and active osteoblasts were found on the surface of the newly formed bone in vivo. CONCLUSION: The complex of PLGA and type-I collagen is an appropriate biodegradable scaffold and can be applied in bone tissue engineering.

Metastatic squamous cell carcinoma of the gingiva appearing as a solitary branchial cyst carcinoma: diagnostic role of PET/CT.

We herein present a case of a left cervical cystic mass, for which the initial pathological diagnosis was branchial cleft cyst carcinoma (following complete mass excision). Thorough postoperative examinations, including with FDG positron emission tomography/computed tomography (PET/CT), revealed a primary tumor in the retromolar region of the left mandible. A 52-year-old female presented with a 2-month history of a painless, progressively enlarged left-sided neck mass. Fine-needle aspiration biopsy suggested a branchial cleft cyst. Physical examination revealed a 3 × 3-cm smooth, tender mass in the upper-left neck and anterior border of the sternocleidomastoid muscle. Examination using nasendoscopy and a strobolaryngoscope revealed no abnormalities of the nasal cavity, nasopharynx, oropharynx, hypopharynx or larynx. MRI of the neck revealed a solitary, round, cystic mass under the left parotid gland. The mass was excised completely. Pathologic results indicated a branchial cleft cyst carcinoma. According to the diagnostic criteria for a branchial cleft cystic carcinoma, PET/CT was performed to detect the occult primary site. PET/CT revealed high FDG uptake in the tooth root of the left mandible. Frozen sections of the mass were indicative of moderate, differentiated squamous cell carcinoma. The carcinoma in the retromolar region of the left mandible was locally excised under general anesthesia. A partial left maxillectomy, partial mandibulectomy, and left radical neck dissection were performed. The patient received postoperative concurrent chemoradiotherapy, and was disease-free at the 8-month follow-up. True branchial cleft cyst carcinoma is rare: once diagnosed, it should be distinguished from metastatic cystic cervical lymph and occult primary carcinoma. FDG PET/CT is useful in the identification of occult primary tumor.

Use of the versatile sternocleidomastoid flap in oral and maxillofacial surgery: our experience.

Our aim was to evaluate the reliability of sternocleidomastoid (SCM) flaps in the reconstruction of defects after oral and maxillofacial resections, and summarise the ways in which morbidity can be reduced. We retrospectively enrolled 65 patients who had malignant tumours resected, and assessed the postoperative viability of the SCM flap. All complications were recorded during a follow up period of 1-64 months. We also investigated the relation between recurrence in regional lymph nodes and their preoperative histological state. The conventional SCM flap, the split SCM flap with only the sternal head, and the SCM flap with a half-thickness clavicular graft, were used to repair different defects. No flaps necrosed completely, and in only 5 cases was there partial loss of the skin paddle. The skin paddle avulsed in 2 cases 2 patients developed wound infections. Only 9 patients developed complications (14%, 9/65). Use of the split SCM flap overcomes the problem of bulk. The combination of the SCM flap and clavicular bone enables early dental implantation. The SCM flap is convenient, reliable, and technically easy for the reconstruction of intraoral or mandibular tissue loss. Preservation of the branch of the superior thyroid artery and precise surgical technique contribute to a higher success rate.

Theoretical and experimental investigation on dissolution and regeneration of cellulose in ionic liquid.

Density functional theory calculations and atoms in molecules theory were performed to investigate the mechanism of cellulose dissolution and regeneration in 1-ethyl-3-methylimidazolium acetate ([emim]Ac), and (1,4)-dimethoxy-ß-D-glucose (Glc) was chosen as the model for cellulose. The theoretical results show that the interaction of [emim]Ac with Glc is stronger than that of Glc with Glc. Further studies indicate that the anion acetate of [emim]Ac forms strong H-bonds with hydroxyl groups of Glc. It is also observed that the H-bonds between [emim]Ac and Glc are weakened or even destroyed by the addition of water. In addition, both the original and regenerated cellulose samples were characterized with FT-IR, XRD, TGA and SEM. The experimental results prove that cellulose can be readily reconstituted from the [emim]Ac-based cellulose solution by the addition of water and the crystalline structure of cellulose is converted to cellulose II from cellulose I in the original cellulose.

Augmentation of transgenic expression by ultrasound­mediated liposome microbubble destruction.

Non-invasive, efficient and tissue-specific transgenic technologies could be valuable in gene therapy. Although non-viral carriers may be safer and cheaper, they have a much lower transfection efficiency than viral gene carriers. The present study was designed to test the transgenic expression and safety of red fluorescent protein (RFP) in HeLa cells in vitro and in transplanted tumors of nude mice in vivo under ultrasound-mediated liposome microbubble destruction (UMLMD) conditions. Plasmids containing RFP were gently mixed with liposome microbubbles (LMs). The mixture was added to HeLa cells or injected into BALB/c mice by the tail vein under various ultrasound exposure and LM parameters, and then the transfection efficiencies were examined. The results in vivo and in vitro demonstrated that, following a comparison of the plasmid group, the ultrasound + plasmid group and the LM + plasmid group, UMLMD significantly increased the transgenic expression (P<0.01) without causing any apparent detrimental effect. From the study, we concluded that UMLMD could be a non-invasive, effective and promising non-viral technique for gene therapy and transgenic research.

Medium-chain fatty acid nanoliposomes for easy energy supply.

OBJECTIVE: Developing a nanoliposome delivery system for an easy energy supply of medium-chain fatty acids (MCFAs) to improve oral doses and bioavailability. METHODS: Bangham's method and high-pressure microfluidization were used to prepare MCFA liposomes. The easy energy-supply property of MCFA nanoliposomes was estimated by the anti-fatigue experiments of mice including a weight-loaded swimming test and its corresponding parameters (serum urea nitrogen, blood lactic acid, and hepatic glycogen). For comparison, nanoliposomes without MCFAs and MCFAs not entrapped in nanoliposomes were used throughout. RESULTS: Compared with crude MCFA liposomes according to Bangham's method, the MCFA nanoliposomes made by high-pressure microfluidization exhibited great advantages in their characteristics, with a small average diameter (76.2 ± 34.7 nm), narrow size distribution (polydispersity index 0.207), high ζ-potential (-50.51 mV), great entrapment efficiency (70.5%) and drug loading (9.4%), and good stability. The high-dose group and the MCFA group (680 mg/kg) showed a longer weight-loaded swimming time (104 ± 29 min, P = 0.087, and 108 ± 11 min, P = 0.047, respectively) and significantly higher hepatic glycogen (16.40 ± 1.45 mg/g, P < 0.001 and 17.27 ± 2.13 mg/g, P < 0.001, respectively) than the control group (59 ± 11 min and 8.79 ± 2.76 mg/g, respectively). Moreover, serum urea nitrogen (891.5 ± 113.4 mg/L, P = 0.024, and 876.6 ± 70.8 mg/L, P = 0.015, respectively) and blood lactic acid (6.05 ± 1.40 mmol/L, P = 0.001, and 5.95 ± 1.27 mmol/L, P < 0.001, respectively) in the high-dose group and the group with an equivalent MCFA dose were significantly lower than those in the control group (1153.6 ± 102.5 mg/L and 12.53 ± 1.86 mmol/L, respectively). CONCLUSION: Similar to MCFAs, MCFA nanoliposomes prepared by high-pressure microfluidization showed a strong easy energy-supply property, which suggested that MCFA nanoliposomes could be a potential drug candidate for an easy energy supply.

[Acupuncture for treatment of kinetic insufficiency of kidney-qi and study on the mechanism].

OBJECTIVE: To observe the therapeutic effect of acupuncture on kinetic insufficiency of kidney-qi and to study on the mechanism. METHODS: Forty-four healthy professional sportsmen were randomly divided into an acupuncture group and a control group. They trained for 4 weeksq with regular detection. In the latter 2 weeks, the acupuncture group received scalp and body-acupuncture. RESULTS: Three weeks later, signs of insufficiency of kidney-qi appeared in most of the sportsmen in the two groups. Two weeks after treatment, in the acupuncture group the signs improved, and testosterone (T) and estradiol (E2) levels increased (P < 0.05); the symptoms did not significantly improve and the T and EZ levels decreased in the control group (P < 0.05). CONCLUSION: Acupuncture therapy can regulate excitatory and inhibitory functions of the nervous system, improve symptoms of kinetic insufficiency of kidney-qi, and has regulative and repairing action on the sexual glands and adrenal gland.

[Biodistribution of (99m)Tc-labeled anti-VEGF mAb 5-FU loaded polylactic acid nanoparticles in human gastric carcinoma xenografts].

OBJECTIVE: To prepare (99m)Tc-labeled Anti-VEGF mAb 5-FU loaded polylactic acid nanoparticles ((99m)Tc-Ab-5-FU-NPs) and investigate its biodistribution in human gastric carcinoma xenografts. METHODS: (99m)Tc-Ab-5-FU-NPs were prepared by labeling Ab-5-FU-NPs with (99m)Tc using improved Schwarz method. After isolation of (99m)Tc-Ab-5-FU-NPs using SephadexG250 column, the labeling ratio and radiochemical purity were determined using chromatography. The immunocompetence of (99m)Tc- Ab-5-FU-NPs was detected by ELISA and immunohistochemistry. (99m)Tc-Ab-5-FU-NPs were then injected via the tail vein into SCID mice bearing human gastric carcinoma, and (99m)Tc labeled mice-derived monoclonal IgG loaded polylactic acid nanoparticles were used as the control, followed by radioimmunoscintigraphic imaging at 2 and 6 h. The radioactive count and radioactive ratio of the tumor and non-tumor tissue (T/NT) in the animal models were calculated using ROI technique. After imaging at 24 h, SCID mice were sacrificed and the radioactive distribution, the %ID/g, as well as the T/NT radioactive ratio were examined, respectively. The concentrations of 5-FU in the tumor and blood were also detected using HPLC method. RESULTS: The labeling ratio of (99m)Tc-Ab-5-FU-NPs was 90%-95%. (99m)Tc-Ab-5-FU-NPs were detected in the tumor tissues by radioimmunoimaging 2 h after the injection. ID%/g in the tumor tissues at 2 and 6 h were both significantly higher than that of the control group. Both the ID%/g in tumor tissues and radioactive ratio of tumor and blood at 6 h were higher than those at 2 h, and the concentration of 5-FU in experimental group increased continuously with time and was significantly higher than that in control group. CONCLUSIONS: (99m)Tc-Ab-5-FU-NPs prepared in this study can meet the demands of radioimmunoimaging, and the anti-VEGF monoclonal antibody possesses reliable immune targeting ability. Six hours after injection, (99m)Tc-Ab-5-FU-NPs can specifically accumulate in the tumor tissues in human gastric carcinoma xenografts at high concentration.

[Study of the anti-tumor effect of anti-vascular endothelial growth factor McAb 5-fluorouracil loaded polylactic acid nanoparticles].

OBJECTIVE: To explore the anti-tumor efficacy of anti- vascular endothelial growth factor (VEGF) McAb 5-fluorouracil (5-FU) loaded polylactic acid (PLA) nanoparticles (NPS) in human gastric carcinoma xenografts of nude mice. METHODS: Anti-VEGF McAb 5-FU loaded PLA NPS were made by ultrasound emulsification. Nude mice model of human gastric carcinoma xenografts was established. Therapeutic effects of drugs on human gastric carcinoma xenografts and side effects concerned were observed. RESULTS: The tumor inhibition rates of control group, nanosphere without 5-FU group, 5-FU (20 mg/kg) group, anti-VEGF McAb nanosphere without 5-FU group, anti-VEGF McAb group, nanosphere with 5-FU group, 5-FU (20 mg/kg) combined with anti-VEGF McAb group, anti-VEGF McAb 5-FU loaded nanosphere group was 0, 6.61%, 24.26%, 27.94%, 35.29%, 37.50%, 39.71% and 52.21% respectively, and there were no significant differences between anti-VEGF McAb 5-FU loaded nanosphere group and nanosphere group without 5-FU in WBC count, serum alanine transferase level or creatinine level. Compared with control group and anti-VEGF McAb 5-FU loaded nanosphere group, the 5-FU group decreased by 34.43% and 37.38% respectively in WBC count (P< 0.05), and increased by 93.17% and 66.56% respectively in alanine transferase. There were significant differences between experimental groups and control group in apoptosis index, especially between anti-VEGF McAb 5-FU loaded nanosphere group and control group (P< 0.05). The microvessel density (MVD) of experimental groups containing anti-VEGF McAb was significantly lower than that of control group or groups containing 5-FU (P< 0.05). CONCLUSION: Anti-VEGF McAb 5-FU loaded nanosphere can increase the tumor inhibitory rate of 5-FU, induce apoptosis by inhibiting tumor angiogenesis with less side effect, and then enhance therapeutic effect, which indicate its potential as a novel, safe nano-tumor-targeting drug.

[Preparation and release efficiency of polylactic acid nanoparticle].

BACKGROUND & OBJECTIVE: As a new drug delivery carrier, medical nanoparticle (NP) appears to be very promising and are widely studied. Compare with microparticle, nanoparticle possesses several advantages, such as ultramicroscopic size, could be ingested by the cells after crossing the tissue matrix, and can penetrate the arterial wall and cross the blood-brain barrier. This study was to prepare polylactic acid (PLA) nanoparticle, and observe its morphology, diameter, structure, surface elements, and ability of in vitro drug release. METHODS: The biodegradable PLA was used as the carrier, and 5-fluorouracil (5-FU) was used as the model drug. 5-FU-PLA nanoparticle (5-FU-PLA-NP) was prepared by matrix and ultrasound emulsification. Morphology of 5-FU-PLA-NP was observed under scanning electron microscope; its surface elements were detected by X ray photoelectron spectroscopyû its drug loading (DL), embedding ratio (ER), and ability of in vitro drug release were assessed by ultraviolet spectroscopy. RESULTS: The nanoparticle was uniformly spherical with average diameter of (191+/-17) nm, DL of 15.2%, and ER of 45.6%. The nanoparticle showed sustained release character in the experiment of in vitro drug release: the cumulative drug release rate in analog body fluid was 94.3% at the 10th day. CONCLUSION: PLA-NP may serve as a carrier of 5-FU, and can change the pharmacokinetics of 5-FU, slower down drug release; 5-FU-PLA-NP can be prepared as intravenous injection, and may prolong the in vivo circulation time of 5-FU, so as to play more efficient antitumor effects.

[Preparation and application of gel chip].

A new method for manufacturing three-dimensional gel film-coated chips was described in this paper and its advantages were evaluated by its application. A patch of polyacrylamide gel (15mm x 15mm x 20 microm) was fixed on the glass surface with Bind-Silane treatment, then activated by glutaraldehyde. The aldehyde groups in gel provided reactive sites that allowed covalent immobilization of molecules containing amino groups. Oligonucleotides were mechanically spotted by GMS 417 Arrayer. After hybridization with Cy-3 labeled probes, fluorescence signals of perfect binding can be discriminated from mismatched ones. Compared with two-dimensional glass chip, the capacity of oligonucleotides immobilized on gel film-coated chip is over 100 times. And the gel film-coated chip have lower background and shorter hybridization time. Monoclonal antibodys of cytokine IL-4, IL-5, IL-6, IL-7, ANG, I-309 and VEGF were also immobilized on the gel film-coated chips to make protein microarrays. After incubation with serum of breast cancer patients or normal persons, the microarray reacted with biotin-labeled second antibodys of cytokines and Cy-3-labeled streptavidin sequentially. Results show IL-4, IL-5, I-309 and VEGF of patients have higher expression level than normal persons. This kind of protein microarrays can be potentially helpful to clinical diagnosis. Furthermore different oligonucleotides or proteins can be performed in parallel in a single reaction with minimal amount of binding reagents. Such gel film-coated chips can be used widely in the fabrication of oligonucleotides and proteins microarrays.