Objectives To establish a model of carotid atherosclerotic (AS)stenosis in rabbits and to preliminarily investigate the expression of asparaginyl endopeptidase. Methods Fourteen New Zealand white rabbits were divided into either an model group (n = 8)or a sham operation group (n = 6)according to the random number table. The carotid intima was injured by operation in the model group. The rabbits in both groups were fed with high fat diets containing magnesium for 10 weeks. The rabbits were weighted and their blood lipids were tested every 2 weeks. At the end of the fifth and tenth weeks after procedure,the plaque and vessel stenosis of the rabbits were observed by MRI. At the end of the tenth week after proce-dure,the specimens were collected and sliced. Hematoxylin and eosin (HE)staining was used to observe the pathological changes. Immunohistochemical staining was used to analyze the expression of asparaginyl endopeptidase (AEP). Results One rabbit in the model group died of carotid artery injury. After being fed with high-fat diets,the body quality and the level of blood lipid were increased in the rabbits of both groups compared with those before procedure (all P < 0. 01). At the end of the fifth and tenth weeks after procedure,MRI revealed that the luminal stenosis rates in the operation group were 16 ± 11% and 53 ± 20% respectively. There was significant difference within the group (t = - 4. 83,P < 0. 01). MRI revealed no luminal stenosis twice in the sham operation group. HE staining showed intimal hyperplasia,AS plaque formation,lipid deposition in plaques,macrophage and smooth muscle cells migration and infiltration forming foam cells in the model group. No AS formation was observed in the sham operation group. The expression of AEP was higher in the rabbit carotid artery tissue in the model group,and it expressed rarely in the sham surgery group. The absorbance values were 0. 072 0 ± 0. 028 0 and 0. 002 0 ± 0. 000 9 respectively. There was significant difference (t = 6. 61,P < 0. 01). Conclusions The methods of injuring carotid intima combined with magnesium containing high-fat diet may exactly,reliably,and quickly establish an AS carotid artery stenosis model. AEP may associat with the occurrence of AS plaques.

OBJECTIVE To study the expression of the level of tumor necrosis factor alpha(TNF-?) and total IgE in Mycoplasma pneumoniae(MP) infection and bronchial asthma,and to analyze its relation with disease severity.METHODS The enzyme-linked immunosorbent assay(ELISA) was adopted to measure the level TNF-? and radioimmunoassay(RIA) to measure IgE in 42 children patients with asthma and MP infection as a test group,and 36 healthy children as a control group.RESULTS Among patients with asthma and MP infection TNF-? [(37.96?9.76)?g/L] and IgE [(310.57?30.39) mg/ml] had a significantly higher level than control group [(16.39?6.87)?g/L and (201.50?16.98)mg/ml],the statistical differences were significant(t=11.23;t=19.56 P

Objective To observe the clinical effect of Renzi Shengjing Decoction(Radix Ginseng and Fructus Schisandrae decoction for producing sperms)treating oligospermia and aspermia males with kidney yang deficiency through its influence on semen and hormones.Methods The 60 males with infertility due to oligospermia and aspermia were randomly divided into treated group and control group with 30 cases in each.Before and after treatment,the two groups were tested with the colored semen analysis system for detecting the density,motility and survival rate of sperms respectively.The follicle-stimulating hormone(FSH),luteinizing hormone(LH),testosterone(T),and prolactin(PRL)were measured by enzyme-labeling instrument.Results In the treated group,the improvement of semen parameters and sexual hormones and clinical effect were better than those of control group(P

Objective To evaluate the effect of Sengstaken-Blakemore tube in assisting cyanoacrylate injection combined with sclerotherapy and variceal ligation under emergency endoscopy in treating escophageal and gastric variceal bleeding. Methods 60 patients were randomly selected, and were evenly divided into 2 groups single blindly. Endoscopic cyanoacrylate injection, sclerotherapy and variceal ligation were performed in control group, and additional Sengstaken-Blakemore tube were used in experimental group. Results All the 30 patients in study group got effective hemostasis (30 of 30, 100.00 %), whereas only 23 patients in control groups stopped bleeding (23 of 30, 76.67 %) (P < 0.05). escophageal and gastric variceal were cured 19 patients in the experimental group (19 of 30, 63.33 %) , and only 4 were cured in the control group (4 of 28, 14.28 %) (P < 0.05). Meanwhile, occurence of fever, chest pain, hospital days and hospitalization costs of experimental group was significantly lower than that in control group (P < 0.05). Conclusions Treating the escophageal and gastric variceal bleeding, using endoscopic cyanoacrylate injection, sclerotherapy; and variceal ligation under emergency endoscopy combined with Sengstaken-Blakemore tube could significantly increase the therapeutic efficiency as well as reduce complications and therapy cost.

Objective To survey and analyze characteristics of brucellosis epidemic in Dalian City for the purpose of setting up prevention and control measures for the disease. Methods In 2007, the basic situation of people of 7-60 years old and in close contact with livestock was surveyed in Dalian according to the "Surveillance standard for brucellosis"(GB 16885-1997). Blood was collected in brucellosis suspicious or high-risk groups for the laboratory examination, using rose bengal test for qualitative detection and test-tube agglutination test(SAT) method for quantitative detection of serum antibodies; At the same time, brucellosis cases found in the routine monitoring and confirmed in and this survey in the department of endemic disease prevention in the center for disease control and prevention of Dalian City underwent questionnaire surveys. Results All 1563 people were epidemiologically surveyed of brucellosis, 1310 were male, 253 female,livestock care givers accounted for 56.05% (876/1563). 240 blood samples were serologically surveyed, the detection rate was 3.75%(9/240). This survey confirmed 2 cases of brucellosis patients, 3 cases confirmed in routine monitoring. 3 cases were infected via contacting infected brucellosis cows, and the 2 cases were imported. Conclusions To strengthen the quarantine of livestock and timely treat the infected livestock should be the key of control of brucellosis. At the same time, professionals of livestock should be educated to protect themselves.

Objective To construct and evaluate a nomogram for predicting the risk of peri-operative complications after radical retro-pubic prostatectomy (RRP) with standard classification criteria.Methods The peri-operative complications and clinicopathological data of 240 patients (50-82 years old) who had undergone RRP for prostate cancer from June 2006 to June 2011 were retrospectively collected.Gleason score:95 cases ＜ 7; 145 patients ≥ 7;Clinical staging:cT1 is 1 case (0.4％),T2a 5 cases (2.1％),T2b 7 cases (2.9％),T2c 162 cases (67.5％),T3a 26 cases (10.8％),T3b 39 cases (16.3％).The peri-operative complications (in 30 days after surgery) were classified by Clavien-Dindo Classification system (the occurrence rates are as follows:rectum injury 1.6％,wound infection 2.0％,deep venous thrombosis 1.2％,urinary leak 5.0％,lymphocele 4.5％,myocardial infarction 5.8％,second look operation 1.6％),and a logistic regression model was used to construct the nomogram.Results BMI,N staging and Blood loss more than 200ml during surgery were independent prognostic factors of RRP morbidity in multivariate logistic regression.The nomogram predicting the risk of peri-operative complications showed relative good concordance index (0.633) and good calibration. Conclusions Based on the clinicopathological factors,a nomogram to predict the probability of peri-operative complications in patients undergone RRP was constructed.This statistical tool may be beneficial in judging operation risk and help consulting with patient before or after surgery.

Objective To study the mechanism of adaptive immunity against Rhizopus arrihizus (R. arrihizus) infections. Methods Bone marrow derived dendritic cells (BMDCs) were separated from C57BL/6 mice and Card9-/- mice and then were cultured in vitro. Resting spores and swollen spores of R. arrihizus were in vitro co-cultured with BMDCs with or without Syk inhibition. Secretion of cytokines ( IL-23, IL-1βand IL-12) was analyzed by ELISA after 24 hours of culture. Na?ve T cells derived from C57BL/6 mice were in vitro co-cultured with spore-stimulated BMDCs for four days. Levels of IL-17A and IFN-γ in supernatants of cell culture were analyzed by ELISA. Flow cytometry was performed to analyze T cell differ-entiation. Confocal microscopy was used to observe the images of stained β-glucan on the surface of resting and swollen spores. Swollen spores were co-cultured with Dectin-1, Dectin-2, TLR2 and mannose receptor ( MMR) , and the binding results were analyzed by flow cytometry. Results Swollen spores but resting spores could induce the maturation of BMDCs and promote the secretion of cytokines (IL-23, IL-1βand IL-12). Co-culturing T cells with swollen spore-stimulated BMDCs enhanced their differentiation to Th17 and Th1. In addition, swollen spores promoted the secretion of Th1-related cytokine ( IFN-γ) and Th17-related cytokine (IL-17A). Adding Syk inhibitor to Card9-/-BMDCs or wild type BMDCs significantly inhibited the secretion of cytokines and T cell differentiation, especially in the Card9-/- group. β-glucan was overserved on the surface of swollen spores, but not on resting spores. On the surface of swollen spores existed pathogen associated molecular patterns ( PAMPs) that could bind with Dectin-1 and TLR2. Conclusion Swollen spores of R. arrihizus could active BMDCs to secrete cytokines of IL-23, IL-1β and IL-12 and trigger T cell responses in vitro. The possible mechanism might be associated with β-glucan exposed on the surface of swollen spores that binds with Dectin-1. The responses between BMDCs and R. arrihizus are Syk-Card9-dependent.

Objective To develop new methods to cultivate, retrieve and purify mouse mesenchymal stem cells (mMSCs). Methods Bone marrow was collected from 2-month-old Kunming mice by flushing femurs and tibias with complete medium of DMEM-LG. Cells were plated in a Petri dish. After 24 hours, non-adherent cells were removed by two to three washes with PBS, adherent cells were further cultured in complete medium and retrieved by trypsinisation with 0.25% trypsin for 5 min at 37 ℃. The treated adherent cells were cultivated with 3?dilution for further generations. CD11b-negative cells were retrieved from the collected adherent cells of 3rd generation by using immunomagnetic microbeads, and continued to be cultured in complete medium. After the cultured cells were retrieved, their morphology and their ability of osteoblastic differentiation and adipocytic differentiation were examined. Results Most of mMSCs from 1st generation were of shuttle shape, some of irregular shape. After treatment with magnetic microbeads and several generations, mMSCs were of spindle, star and irregular shape. These cells were of rich cytoplasma, clear nucleolus, and grew in parallel or vortex. The cultured adherent cells from the first and subsequent generations had plenty of CD11b-positive blooding-making cells. After 20-day osteoblastic induction, mMSCs differentiated into bone cells, which showed orange phosphate in extracellular matrix by Alizarin red S staining. mMSCs could differentiate into lipocytes. The size of cells increased along with fat-developing induction period. These cells showed many orange fatty follicles with O Red Oil dyeing. Conclusion Pure mMSCs can not be retrieved by either adhering method or generation cultivation method separately. The combined methods of adhering, immunomagnetic microbeads, and serial subcultivation is effective in vitro in retrieve mMSCs.

Objective: To observe the effect of 2 Hz electroacupuncture (EA) on long term depression (LTD) of synaptic transmission in the spinal dorsal horn in rats with neuropathic pain, so as to explore the central mechanisms of the antinociceptive effects of 2 Hz electroacupuncture on neuropathic pain. Methods: The neuropathic pain models were produced by tight ligation of the L5/L6 spinal nerves in Sprague Dawley rats. The C fiber evoked field potentials in the spinal dorsal horn were recorded with extracellular recording techniques. The parameters of the electroacupuncture were as follows: frequency of 2 Hz, wavelength of 0.6 ms, intensity of 1,2,3 mA lasting 10 min for each intensity, stimulation time of 30 min. The positive stimulating electrode was placed in acupoint “sanyinjiao” and the negative electrode in “zusanli”. Results: (1) 2 Hz electroacupuncture significantly decreased the amplitudes of C fiber evoked field potentials in the spinal dorsal horn in rats with neuropathic pain to (49.4?0.6)% of the control, compared with that (100.1?1.2) % of the control before EA (unpaired t test, P

BACKGROUND:Adipose-derived stem cels are regarded as the potential seed cels for tissue engineering. Colagenase digestion is used to isolate adipose-derived stem cels from fat pads currently. However, there are some problems, such as cumbersome operation and high cost.
OBJECTIVE: To study the basic biological characteristics of adipose-derived stem cels by tissue explants culture and to explore the differentiation potential into osteoblasts, adipocytes and endothelial progenitor cels in vitro.
METHODS:Adipose-derived stem cels were isolated by tissue explants technique from the bilateral groin fat pads of rats under aseptic conditions, and cultured in vitro. Cel counting kit-8 was used to detect the proliferative activity, and flow cytometry was employed to analyze the expression of cel surface markers. Passage 4 adipose-derived stem cels were cultured in osteogenic medium, adipogenic medium and endothelial progenitor cel medium for 2-3 weeks, and then the cels were identified.
RESULTS AND CONCLUSION:Adipose-derived stem cels that were isolated by tissue explants culture were easily cultured, and after subculture, cels were mainly spindle-shaped and grew in clone-like manner with swirling arrangement. Cels that experienced repeated subcultures stil kept stronger proliferative ability and the cel growth curve was shaped as a parabola. Immunochemical staining analysis revealed that adipose-derived stem cels were positive for CD44, CD90 and CD29, but negative for CD31, CD45. After adipogenic/osteogenic induction, the cels were respectively positive for oil red O staining and alizarin red staining. Induced endothelial progenitor cels were identified with CD34 and the ability to uptake Dil-ac-LDL and FITC-UEA. These findings indicate using the using tissue explants culture, high-purity adipose-derived stem cels easy to proliferate can be harvested, highly express stem cels-related antigens, and have the ability to differentiate into osteoblasts, adipocytes and endothelial progenitor cels, which meet the needs of seed cels in tissue engineering research.