Objective To construct the prokaryotic expression vector for HD-5 and purify the recombinant HD-5 protein then analyze its antifungal activity. Methods The HD-5 gene was cloned by PCR, then was inserted into prokary-otic expression plasmid pQE-30Xa to construct pQE-30Xa/HD-5. After sequencing, pQE-30Xa/HD-5 was transformed in-to E.coli M15 cells. Its expression was induced by IPTG and confirmed by SDS-PAGE. The recombinant protein was purified through Ni-NTA affinity purification system. The antifungal activity was tested by disk diffusion method. Results HD-5 gene and pQE-30Xa/HD-5 vector were obtained successfully. E.coli M15 strains was used to express HD-5 fusion protein. After purification, the fusion protein was confirmed by Western blot. The disk diffusion test confirmed that the fusion pro-tein can inhibit Candida albicans. Conclusion Expression vector pQE-30Xa/HD-5 was successfully constructed. The HD-5 fusion protein was expressed in E.coli successfully, which showed a certain degree of antifungal activity.

Objective To study the effects of antimicrobial peptide LL-37 expressed and purified from prokaryotes on candida albicans growth. Methods (1) Thirty female Kunming mice were treated with estrogen and white candida yeast suspension were poured into vagina to establish a vulvovaginal candidiasis (VVC) murine model. After successful establishing the VVC mouse model, mice were randomly sorted into test group (n=15) and control group (n=15) . Suspension(30μl, 100μg/ml)of recombinant peptide LL-37 expressed and purified in Prokaryotes was given by intravaginal administration to the test group for 5 days, while the same amount of phosphate buffered saline (PBS) was given to the control group. (2) Tweenty-four hours after treatment, the fungal burden and colony-forming unit (CFU) of vaginal fluids were evaluated. All mice were subsequently sacrificed and vaginal tissues were harvested for tissue homogenate preparation. ELISA was used to determine the levels of nterleukin-10(IL-10)and interferon-γ(IFN-γ)in the isolated vaginal tissues. Results (1) VVC mouse model was established successfully in this study. Vaginal mucosa congestion, edema, vaginal plica disappearing were obviously observed in the control group. After treatment with recombinant protein LL-37 vaginal mucosa has no obvious change in the test group. (2) Fungal burden and CFU of vaginal fluids were significantly lower in the test group [(4.8±1.0)×104 CFU/ml] than that in the control group [(8.5 ± 2.1) × 104 CFU/ml, P=0.017]. IFN-γlevel of the test group was increased [(257 ± 11) vs (197 ± 4) pg/ml, P=0.000], while the level of IL-10 was reduced [ (287 ± 15) vs (379 ± 17) pg/ml P=0.000] resulting in a the ratio of IFN-γ/IL-10 was in significantly higher in test group (0.892±0.008 vs 0.496±0.013, P=0.000). Conclusion Recombinant protein LL-37 expressed and purified from prokaryotes inhibits the growth candida albicans and improves vaginal immunity by adjusting IFN-γand IL-10 secretion in the VVC mouse model, highlighting the therapeutic potential of LL-37 for VVC.

Objective To evaluate the changes of the upper airway of the patients with obstructive sleep apnea syndrome (OSAS) before and after operations and to know the effects of operations by MSCT. Methods The upper airway dimensions of 26 patients with OSAS were measured on multiplanar reformatted (MPR), curved-planar reformatted (CPR), volume rendering(VR) images of 16-slice spiral CT. The measurements include the anteroposterior calibres and the areas on the reformatted axial images on the pharyngeal cavity levels, the calibres and the minimum areas in retropalatal and retroglossal regions, the areas of the soft palate and uvula on the reformatted sagittal view with maximum thickness, the maximum wall thickness of the right and left the upper airway on the coronary images, the volume of the upper airway before and after the operations. The measurements were correlated with the polysomnography (PSG) records. The data were analyzed paired-samples t-test and Pearson correlations. Results By comparison, the anteroposterior calibres and the cross-sectional areas on the reformatted axial view of the lower retropalatal region (slice 4) of the upper airway increased significantly after operations. The anteroposterior diameter increased from 5. 9 mm before operations to 12.8 mm after operations, where t = - 5.506, P < 0.05. The areas increased from 51.0 mm~2 before operations to 275.0 mm~2 after operations, where t = -5.011, P <0.05. In the higher retropalatal region (shce 2) of the upper airway, the anteroposterior diameter decresased from 14.8 mm before operations to 9.2 mm after operations, where t = 2.867, P < 0.05. The areas decreased from 241.0 mm~2 before operations to 128.0 mm~2 after operations, where t = 3.087, P < 0.05. The anteroposterior calibres of retroglossal region (slice 7) decreased from 12.7 mm before operations to 10.3 mm after operations,where t = 3.718, P <0.05. The L-R calibres and the minimum areas of of retropalatal increased significantly from 6.4 mm, 33.0 mm~2 before operation to 10.9 mm, 76. 0 mm~2 after operation, where t = -3.413, -2. 216, respectively and P < 0.05. Of the 9 cases whose apnea and hypopnea index (AHI) ≤5 events/hour after operations, the minimum areas of retropalatal region, the anterio-posterior diameter, L-R calibres increased significantly. The areas increased from 41.0 mm2 before operations to 76.0 mm~2 after operations, were t = -4. 932, P <0.05. The anteroposterior calibres increased from 4.6 mm before operations to 6.6 mm after operations, where t = - 7. 308, P < 0.05. The L-R calibres increased from 8.3 mm before operations to 13.6 mm after operations, where t = - 4.320, P < 0.05. Conclusions MPR、CPR、VR of MSCT can evaluate the not only the morphology but the function changes of the upper airways on the OSAS patients. The increasing of the minimum cross-sectional area may be one of the important indications for evaluating operations. The narrowing of the higher retropalatal region of the upper airway after operations should be an alert to the clinicians.

Objective To study the inhibitory effect of antimicrobial peptide LL-37 on Candida albicans through its ability to promote the secretion of immune factors by vaginal epithelial cells. Methods (1) LL-37 prokaryotic expression vector pET-Duet/LL-37 was constructed and its expression was induced in Escherichia coli M15. The expressed LL-37 fusion protein was purified and identified by western blot. Antifungal activity of the purified protein was initially identified by Kirby-Bauer (K-B) method. (2) Purified LL-37 protein was added to human vaginal epithelial cells co-cultured with Candida, and inhibitory effect on Candida growth was determined by the glucose consumption method. Interferon γ (IFN-γ), interleukin 10 (IL-10) concentration and IFN-γ/IL-10 ratio were measured by ELISA at different time points. Results (1) LL-37 fusion protein was purified to 96% purity at a concentration of 433.92 μg/ml, and was shown to possess anti-fungal activity confirmed by the K-B method. (2) A Candida-vaginal epithelial cells co-culture system was successfully constructed. LL-37 recombinant protein inhibited the growth of Candida with absorbance values significantly higher in the treatment group compared to the control group at all measured time points (12-hour:3.008±0.003 versus 2.967±0.003, 24-hour:2.941±0.003 versus 2.601±0.003, 48-hour:2.893 ± 0.004 versus 2.409 ± 0.003; all P<0.01). Furthermore, the rate of decrease was also much slower compared to the control group. In both control and experimental groups, IFN-γ and IL-10 secretion levels were observed to rise at first peaking at 24 hours and subsequently decrease. For each time period, IFN-γconcentration in the experimental group was significantly higher at 24 hours compared to the control group [(104.00 ± 1.07) versus (85.17 ± 0.28) pg/ml, P<0.01]. In contrast, IL-10 concentrations were significantly lower than the control group at all time points (P<0.01). IFN-γ/IL-10 ratio was also observed to be significantly higher than the control group at all measured time points (P<0.01). Conclusions (1) Recombinant protein LL-37 could significantly inhibit the growth of Candida. (2) By influencing the secretion of immune factors such as IFN-γ, IL-10, etc, recombinant protein LL-37 is able to adjust vaginal epithelial cells local immunity, and enhance resistance to Candida infection.

OBJECTIVE To study the isolation and resistance tendency of Acinetobacter calcoaceticus-baumannii to antimicrobial agents from 1998 to 2004 to provide valuable data for infection prevention and therapy. METHODS We reviewed the isolation rates,distribution in clinical specimens and wards,and the resistance rates of(A.calcoaceticus-baumannii)to 14 kinds of antimicrobial agents from 1998 to 2004. RESULTS There was an increasing tendency of isolation rates of A.calcoaceticus-baumannii every year,which was 0.18% in 1998 but 1.48% in 2004.In the seven years,there was the highest isolation rate of 70.58% in specimens from respiratory tract,the next was from the urine(9.42%),and blood(4.63%).Concerning the wards distribution,ICU had the highest rate of 47.28%.In 1998,A.calcoaceticus-baumannii had resistance rates more than 50% only to one kind of antimicrobial agents(aztreonam),but in 2004,it had increased to thirteen kinds(except cefoperazone/sulbactam).About the fourteen kinds of antimicrobial agents we inspected,that were increased in their resistance rate.The highest increasing of resistance rate was ceftazidime from 11.1% in 1998 to 88.9% in 2004,the imipenem was second for 0.0% to 64.8%,and the third was sulfamethoxazole/trimethoprim form 0.0% to 64.0%,while there still was an increasing resistance tendency to them. CONCLUSIONS The clinical isolation rate of A.calcoaceticus-baumannii is increasing,and it has higher resistance rates to many antimicrobial agents as well as an increasing resistance tendency to relatively susceptive antimicrobial agents every year.So physicians should prescribe on the basis of antimicrobial agents susceptibility tests in vitro.

BACKGROUNDRheumatoid arthritis (RA) is a chronic, systemic autoimmune inflammatory disorder. Many methods have been used to observe the progress of RA. The purpose of this study was to observe the progress of RA in rats with 18F-fluorodeoxyglucose (FDG) positron emission tomography/computed tomography (PET/CT), magnetic resonance (MR) imaging and arthritis score, and analyze the relationships among different methods in evaluation of RA.

METHODSSixteen healthy Sprague Dawley (SD) rats about 8-week old were randomly assigned to a RA group and a control group. Bovine type II emulsified incomplete Freud's adjuvant was used to induce arthritis in the RA group. Arthritis score of the rats in two groups were recorded, and (18)F-FDG PET/CT, MR imaging were performed both on the corresponding rats every 3 days. All the rats were sacrificed at week 5, and histopathological examination was performed on rat knees stained with haematoxylin and eosin.

RESULTSThe arthritis score and the standard uptake value (SUV) of knee joints in RA rats increased with the progression of arthritis gradually. Both peaks of arthritis score and SUV appeared at 21 days after the first immune injection, then the arthritis score and SUV of knee joints decreased slowly. The arthritis scores of knee joints in RA rats were positively correlated with their SUV changes. The MR images were confirmed by the histopathological studies.

CONCLUSIONPET/CT can detect the earliest molecular metabolism changes of RA, and MR imaging can follow up the dynamical anatomical changes of RA, all of which indicated that PET/CT and MR imaging may be applied as useful tools to monitor the progress of RA.

Animals ; Arthritis, Rheumatoid ; diagnosis ; pathology ; Fluorodeoxyglucose F18 ; Magnetic Resonance Imaging ; Positron-Emission Tomography ; Radiopharmaceuticals ; Rats ; Rats, Sprague-Dawley ; Tomography, X-Ray Computed

OBJECTIVETo investigate the acquaintance to orthodontics and possible factors that obstruct orthodontic treatment in middle school students in zones with different economic levels in Jiangjin, Chongqing.

METHODSThe students were randomly divided into four groups: A class from a junior high school in town (group 1), a class from a senior high school in town (group 2), a class from a junior high school in the countryside (group 3), a class from a senior high school in the countryside (group 4). The information was collected by questionnaire, and the statistical analysis of results was carried out by SPSS 17.0 software.

RESULTSSex made difference in the acquaintance to orthodontics treatment. Girls worried more about teeth extraction and pain that may occur during treatment than the boys. Students in the city had more access to related consultations compared to those in the countryside. Different economic conditions made much contribution to the proportion to go for orthodontics treatment in cities and rural areas. The evaluation from families and friends may affect patients' subjective estimation to themselves.

CONCLUSIONOrthodontists should learn more about student patiens from various aspects, and make different therapies according to their sexes, economic conditions of family, education levels, etc. Only so can orthodontists get better compliance from their patients.

Adolescent ; Female ; Humans ; Male ; Orthodontics ; Students

Objective To investigate the effects of quercetin with a special focus on their effect on macrophage po-larization after SCI.Methods Adult C57 mice underwent T10 spinal cord clip compression injury,then were ran-domly divided into SCI group and quercetin group.1-14 d after SCI received quercetin by intraperitoneal injection(once a day)in quercetin group,and received same normal saline in the SCI group.3,7 and 14 d after SCI, the lesion section of SCI group and Quercetin group stained by immunofluorescent:M1 macrophages pheno-type cell labeled with iNOS, and M2 macrophages phenotype cell labeled with Arg1.The mRNA expression of inflammatory factors in lesion site was detected by RT-PCR.The motor function was evaluated by Basso mouse scale(BMS)after SCI.Results Compare with SCI group,the expression of arginase-1(associated with M2 mac-rophage phenotype)significantly increased in quercetin group(P<0.05).The expression of inducible nitric oxide synthase-iNOS(M1 phenotype marker)was down-regulated as demonstrated using immunohisto-chemistry(P<0.05).Furthermore,the production of NOS2 and tumor necrosis factor alpha was significantly reduced whereas the level of interleukin 10 and TGF-β were elevated in quercetin group(P<0.05).The time course of functional recov-ery revealed a gradual recovery in the subacute phase in quercetin group, little improvement was observed in SCI group(P<0.05).Conclusions It is found that quercetin may promote the shift of M1 to M2 phenotype and amelio-rate the inflammatory microenvironment.Furthermore,the roles of quercetin in immunity modulation may enhance neuroprotective effects and partially contribute to the locomotor functional recovery after SCI.

Objective: Whether metabolic redistribution occurs in patients with white matter hyperintensities (WMHs) on magnetic resonance imaging (MRI) is unknown. This study aimed 1) to propose a measure of the brain metabolic network for an individual patient and preliminarily apply it to identify impaired metabolic networks in patients with WMHs, and 2) to explore the clinical and imaging features of metabolic redistribution in patients with WMHs. Materials and Methods: This study included 50 patients with WMHs and 70 healthy controls (HCs) who underwent 18F-fluorodeoxyglucose-positron emission tomography/MRI. Various global property parameters according to graph theory and an individual parameter of brain metabolic network called “individual contribution index” were obtained. Parameter values were compared between the WMH and HC groups. The performance of the parameters in discriminating between the two groups was assessed using the area under the receiver operating characteristic curve (AUC). The correlation between the individual contribution index and Fazekas score was assessed, and the interaction between age and individual contribution index was determined. A generalized linear model was fitted with the individual contribution index as the dependent variable and the mean standardized uptake value (SUVmean) of nodes in the whole-brain network or seven classic functional networks as independent variables to determine their association. Results: The means ± standard deviations of the individual contribution index were (0.697 ± 10.9) x 10-3 and (0.0967 ± 0.0545) x 10-3 in the WMH and HC groups, respectively (p < 0.001). The AUC of the individual contribution index was 0.864 (95% confidence interval, 0.785–0.943). A positive correlation was identified between the individual contribution index and the Fazekas scores in patients with WMHs (r = 0.57, p < 0.001). Age and individual contribution index demonstrated a significant interaction effect on the Fazekas score. A significant direct association was observed between the individual contribution index and the SUVmean of the limbic network (p < 0.001). Conclusion The individual contribution index may demonstrate the redistribution of the brain metabolic network in patients with WMHs.

In the fifth scientific and technological revolution， information technology is the first productivity， which has a great impact on the supply and demand of medical services. Generally， internet medicine is equivalent to the combination between health industry and information technology. "Healthy China" strategy is China's "priority development strategy"， which adheres to the principle of health equity， emphasizes the integration of health into all policies， and takes co-construction and sharing as the basic path. With implementing internet medicine， "Healthy China" strategy promotes the mobility of medical service with three tools： interconnection， data and artificial intelligence. This enhances the operation efficiency of overall medical and health system， and optimizes the allocation of medical resources. The future development of internet medicine follows the double helix mode driven by technology and policy， and the policy determines the development boundary of the industry. On the basis to ensure medical safety， we should explore the possibility of internet diagnosis and treatment further， and pay attention to the fairness of resource allocation while improving efficiency， so as to realize the co-construction and sharing of health services.