Objective To localize Der f 2 in the body of Dermatophagoides farinae.Methods Live mites were embedded.Serial mite paraffin sections were made and checked under light microscope.The sections were incubated with anti-recombinant Der f 2 monoclonal antibody as the first antibody.They were then reacted with the fluorescent isothiocyanate conjugated goat anti-mouse IgG as the secondary antibody.The sections were examined with fluorescent light microscopy.Results Der f 2 reacting with immunofluorescent antibodies was found localized in the tissue and contents of the mid-gut of the mite.Conclusion The major allergen Der f 2 distributes in the gut and fecal pellets of Dermatophagoides farinae.

AIM: To establish an apoptotic model induced by 1-methyl-4-phenylpyridinium ion(MPP~+) in PC12 cell and assess the effect of Semen Cuscuta extracts on protecting PC12 cell from apoptosis induced by MPP~+. METHODS: The cell viability was analyzed by MTT method,the morphological changes were observed by Hoechst 33258 staining assay,and the apoptosis rate was measured by flow cytometry analysis(FCM).(RESULTS:) Treatment of 0.15 mM MPP~+ for 48 h induced apoptosis in PC12 cells;concentration of 50 mg/L Semen Cuscuta extracts could protect cells from apoptosis induced by MPP~+. CONCLUSION: MPP~+-induced apoptosis in PC12 cell is greatly inhibited by Semen Cuscuta extracts.

Objective To observe the process of Vibrio vulnificus inducing dendritic cell strain apoptosis.Methods We established the mixed culture model of mouse dendritic cell ( DC 2.4 strain) and Vibrio vulnificus( Vv1.1758 strain ),analyzed morphological characteristics of cell apoptosis by DAPI fluorescence staining,detect DNA fragmentation level of apoptosis cells by DNA Ladder assay,analyze DC2.4 apoptosis rate by Annexin V FITC/PI staining,determine activities of caspase-3 and caspase-8 by means of spectrophotometric method and detect changes of mitochondrial transmembrane potential ( △ Ψm ) by JC-1 fluorescence labeling.Results After Vv1.1758 strain and DC2.4 cell were mixed and cultured for 4 h,DAPI fluorescence staining showed typical apoptosis characteristics-chromatin condensation and marginalization; DNA agarose gel electrophoresis showed apoptosis band; apoptosis rates at 2,4 and 6 h were respectively (37.8±9.8) ％,(54.3 ± 12.7 ) ％ and ( 68.2± 14.6 ) ％ ; Mitochondrial transmembrane potentials (△Ψm) at 1 h,2 h and 4 h reduced by 7.1％,16.1％ and 46.7％ respectively; caspase-8 activity increased at 1.5 h and reached the peak at 2 h [ (2.48±0.19) U/μg],while caspase-3 activity started to increase at 3h and reached the peak at 4 h [ ( 1.91 ±0.16) U/μg ].Conclusion Vibrio vulnificus could induce dendritic cells by two pathways: reducing mitochondrial transmembrane potential and activating caspase-8 promoter and finally activate effector caspase-3 to promote apoptosis.

Objective To investigate the invasion,internalization and the organelles damage of the cultured dendritic cells ( DC2.4 strain) during Vibrio vulnificus (Vv) infection.Methods The study model was the cultured DCs infected by Vv 1.1758 strain.Electron microscopy was used to observe the localization of bacteria in different time point of infection,cell morphology and the process of organelles changes.The cytoskeleton structure including the microfilaments and the microtubules rearrangement was examined by the fluorescence microscope.Results The Vv were pinocytosed into the DC cells through double-sides,and localized at 1-2 μm of the inner side membrane.It cost 1.27,1.87,3.43 hours reaching the infection ratio of 25％,50％,75％,respectively.Using electron microscopy,the DCs had been observed the phagosome formation within 1h,chromatin activation within 2 h,chromatin aggregation 4 h,and the significant cytoskeleton structure disruption within 6 h.Endoplasmic reticulum,mitochondria and lysosomes became swollen.In DCs,the protruding filaments gradually reduced,and their shape changed from the point-like to the linearlike aggregation at the inner side of the plasma membrane,extended microtubules disappeared,the microtubules at the outside nuclear membrane striking rearranged.Conclusion After DC was infected by Vv,the bacteria were pinocytosed into the inner side of DC membrane,and the microfilaments were observed to move from the cytoplasm to cell membrane.In addition,the microtubules moved from the synapse and the cell membrane to the nuclear membrane.The high lethality of Vv could provoke to the DCs cytoskeleton rearrangements.

BACKGROUND:The phenomenon of osteoinduction by biomaterials has been proven in animal experiments. OBJECTIVE:To investigate whether the ability of a biomaterial to initiate bone formation in ectopic implantation sites improves the performance of osteoinductive biomaterial as a scaffold for tissue-engineered bone. METHODS:We compared ectopic bone formation by combining autologous adipose-derived stromal cells with an osteoinductive and a nonosteoinductive biphasic calcium phosphate ceramic to create a tissue engineering construction in the muscle of dogs. Al implants were implanted in the back muscle of 10 adult dogs for 8 weeks and 12 weeks, including osteoinductive biphasic calcium phosphate ceramic+adipose-derived stromal cells (osteoinductive complex group), osteoinductive biphasic calcium phosphate ceramic (osteoinductive broup), nonosteoinductive biphasic calcium phosphate ceramic+adipose-derived stromal cells (nonosteoinductive complex group), and nonosteoinductive biphasic calcium phosphate ceramic (nonosteoinductive group). Micro-CT analysis and histomorphometry were performed to evaluate and quantify ectopic bone formation. RESULTS AND CONCLUSION:Ectopic bone formation was visible in the osteoinductive complex group and osteoinductive group, and the former group was superior to the latter one in quality of new bone (P<0.05). However, there was no ectopic bone formation in the other two groups. Micro-CT results were consistent with the histomorphological detection. These findings indicate that osteoinductive biphasic calcium phosphate ceramic, as a kind of bone tissue engineering scaffold material, has a better osteogenic capacity, while adipose-derived mesenchymal stem cells serve as seed cells to promote the ectopic bone formation.

Objective To analyze the association of blood uric acid level with the severity of coronary artery stenotic changes, metabolic syndrome (MS), and its components. Methods A total of 343 individuals ( male 223,female 120) who underwent coronary angiography and had complete data on MS and serum uric acid were collected. The severity of coronary artery disease (CAD) was assessed by the coronary stenesis index (CSI). MS was diagnosed according to the Guideline on Prevention and Treatment of Blood Lipid Abnormality in Chinese Adults. Results (1)The mean uric acid level was significantly lower in women than in men [ ( 306.3±76.9 vs 358.9±85.2 ) μmol/L, P＜0.01 ]. The prevalence of MS and its components showed no difference between men and women. (2) The uric acid level in women with 3 components was higher than those with1( P＜0. 01 ) or 2 ( P＜0.05 ) components of metabolic disorders, but not in men. (3) Quartiles of concentration of uric acid were computed. Compared with those in the lowest quartile of uric acid, women in the highest quartile had higher CSI score [ 7.0 (2.5-12.0) vs 2. 0( 0.0-6.0), P= 0. 025 ]. Moreover, the uric acid level was higher in women with multivessel lesions than nonCAD patients [ (327.0±81.9 vs 284.9±78.6) μmol/L, P = 0.033 ]. However, no correlation was found between uric acid level and the severity of coronary artery lesion in men. (4) Logistic regression showed that age (β=0.042, P=0. 007) and dyslipidemia(β=0.836, P=0. 037 ) were the independent risk factors of CAD in men, and hypertension(β=1. 127, P=0.039) and dyslipidemia(β=0.901, P=0.009)in women. Conclusions In women with higher uric acid level, the clustering of metabolic abnormalities was increased, and the coronary artery lesion was more severe. High uric acid level might be a marker of CAD for women.

Objective To analyze the clinical characteristics of coronary artery disease (CAD) in patients with nonalcoholic fatty liver disease ( NAFLD). Methods Totally 234 subjects underwent coronary angiography, including 148 men and 86 women with complete data on metabolic syndrome ( MS) and abdominal ultrasonography; the mean age was 66. 6 years. The severity of CAD was assessed by coronary stenosis index (CSI). Metabolic syndrome was diagnosed according to the Guideline on Prevention and Treatment of Blood lipid Abnormality in Chinese Adults. Results Ultrasonography revealed that 62 patients had NAFLD (26. 5%). In patients with NAFLD, the prevalence of central obesity was higher than those without NAFLD (75. 8% vs 50. 0% , P< 0.01). With regards to age, CAD patients with NAFLD were more common in patients under 60 years (27.4% vs 13.7% , P=0.005). CSI score was similar in CAD subgroup and CAD & NAFLD subgroup (P>0.05), however the age of patients in CAD & NAFLD subgroup was significantly lower compared to CAD subgroup ( P = 0.006). According to the results of logistic regression, central obesity was the independent risk factor of NAFLD (β= 1.701, P<0.001). Logistic regression demonstrated that age was independently associated with CAD (β = 0.032, P=0.027). Further more, multiple stepwise regression analysis showed that age was the single parameter that best predicted CSI score (β= 0. 125, P = 0. 022). Conclusions It is important to screen coronary artery disease in middle aged patients with central obesity or NAFLD.

As a pivotal part of the elbow joint structure,coronoid process of the ulna plays a vital role in maintaining stability of the elbow joint.Loss of coronoid process height probably causes instability of the elbow joint.Diagnosis and treatment of coronoid process fracture is a widespread concern for orthopaedic surgeons.Nevertheless,reconstruction of the defects following coronoid process fracture has been rarely reported though they affect elbow stability.It is very challenging to treat the elbow instability induced by coronoid process defects,mostly complicated with other elbow injuries.Moreover,the clinical treatment efficacy remains elusive.This review summarizes the research advances in reconstruction of coronoid process defects.

BACKGROUND:Chronic visceral pain is one of major complaints of irritable bowel syndrome which seriously affects patient’s quality of life. Recent researches have shown that moxibustion therapy has positive effects on aleviating chronic visceral pain in irritable bowel syndrome patients.
OBJECTIVE: To study the clinical utility of moxibustion in coping with chronic visceral pain of irritable bowel syndrome patients, and shed light on the theoretical basis of moxibustion analgesia, thereby to give insights into the further research and application on moxibustion.
METHODS: With the key words of “moxibustion, irritable bowel syndrome, visceral pain, abdominal pain” in Chinese and in English, respectively, a computer-based search was performed in CNKI, VIP, Wanfang and PubMed databases for articles published from January 1990 to October 2014. After the initial screening, the remained articles went through further selection and categorization.
RESULTS AND CONCLUSION:The result shows promising results of moxibustion on relieving chronic visceral pain for both two subtypes of irritable bowel syndrome patients, diarrhea type and constipation type. Moxibustion may exert an analgesic effect on chronic visceral pain in irritable bowel syndrome patients through regulation of visceral hypersensitivity, gastrointestinal motility disorders, brain-gut axis and neuroendocrine system disorders, immune dysfunction and low-grade inflammation in the gut, psychological abnormalities, and alteration of intestinal flora. However, to fuly understand the analgesia effect of moxibustion and elucidate its mechanism, more standardized randomized controled trials employing advanced scientific techniques and equipments wil stil be needed in the future.

Objective To obverse the changes ofmacular choroidal thickness (CT) in patients with mild to moderate Alzheimer's disease (AD).Methods This was a case-control study.Twenty-one patients with mild to moderate AD confirmed by Neurology Department of Jinhua Central Hospital from November 2016 to June 2018 and 21 age-matched control subjects were concluded in the study.There was no significant difference in age (t=0.128),intraocular pressure (t=0.440) and axial length (t=1.202) between the two groups (P＞0.05).There was significant difference in mini-mental state examination score (t=8.608,P＜0.05).CT was measured by OCT with enhanced depth imaging technique in the subfoveal choroid,at 0.5 mm and 1.0 mm from the center of the fovea nasal (NCT0.5,1.0mm),temporal (TCT0.5,1.0mm),superior (SCT0.5,1.0mm),and inferior (ICT0.5,1.0mm).Independent-samples t test was used to compare the results obtained from these two groups.Results SFCT (t=2.431),NCT0.5,1.0 mm (t=3.341,2.640),TCT0.5,1.0 mm (t=3.340,2.899),SCT0.5.1.0 mm (t=3.576,3.751) and ICT0.5 (t=2.897,2.903) were significantly thinner in AD eyes than those in control eyes.Conclusion Compared with healthy subjects,patients with mild to moderate AD showed a significant reduction in CT.