Objective To compare the immune regulation of syngenic and allogenic mesenchymal stem cells (MSCs) in the transplantation combined with islets. Methods After induction of diabetes in 30 Lewis rats with streptozotocin (STZ), the recipient Lewis rats received islets from SD rats combined with syngenic (group B) or allogenic (group C) MSCs injection via the portal vein. The group of islets transplanted alone served as control (group A). The survival time of grafts in all groups was assessed by the level of blood glucose. ELISA was used to detect the levels of interferon-γ (IFN-γ), interleukin 2 (IL-2), IL-4 and IL-10 in the peripheral blood on the 1st and 3rd day after transplantation. Results The blood glucose levels in all three groups were decreased in a normal range (13. 9 mmol/L) and the survival time of grafts in group B (11.38 ± 4. 03 days) was significantly longer than in group C (5. 50± 2. 07 days) as well as group A (2. 88 ± 1.25 days). On the 1 st and 3rd day after transplantation, the levels of TH 1 cytokines IFN-γ and IL-2 in group A were significantly higher than in groups C and B (P＜0.05). Meanwhile the levels of TH 2 cytokine IL-10were increased in group B, but there was no significant difference between groups A and C (P＞0.05). The levels of IL-4 had no significant difference among these three groups (P ＞ 0.05).Conclusion Islet transplantation combined with MSCs could prolong the survival time of grafts.Syngenic MSCs, superior to allogenic ones, were more effective in changing the balance of TH1/TH2to TH2. Decreased expression of TH1 cytokine (IFN-γ, IL-2), which was closely related to the induction of immune tolerance, was beneficial to the long-term survival of grafts.

Objective To establish a HPLC method for the determination of quercetin from different parts of L. cuncata(Dum. Cours.)G.Don.,including roots,old branches,young shoots,leaves and seeds,and to build the fingerprints. Methods The HPLC method determination of quercetin and fingerpints were establised. Results The restults showed that there were great differences be?tween the quercetin contents from different parts,with the highest contents found in seeds,followed by young shoots,leaves,old branches,and roots. The similarities of HPLC fingerprints of the medicinal material were quite different from the parts of L. cuncata (Dum.Cours.)G.Don.. The similarities were all above 0.95 for L. cuncata(Dum.Cours.)G.Don. samples,roots,old branches and young shoots below 0.85,leaves and seeds similarities below 0.60. Conclusion It was concluded that different parts(such as roots, old branches,young shoots,leaves,seeds)of L. cuncata(Dum.Cours.)G.Don. should be divided in clinical and productive practice so as to supply the scientific basis for enhancing the curative effect and reasonable utilization of the resource of L. cuncata(Dum.Cours.)G. Don.

Objective To observe the role of nitric oxide(NO) in the immune regulatory effect of bone marrow mesenchymal stem cells(MSCs). Methods Bone marrow MSCs were isolated from Lewis rats by adherence screening. Concanavalin A (ConA) was adopted as the stimulator and T-lymphocyte isolated from peripheral blood of SD rats as the reactive cells. The changes of the ability of T-lymphocyte proliferation, when co-cultured with MSCs, were measured by CCK-8 assay. The inducible nitric oxide synthase (iNOS) mRNA and protein expression on MSCs and were detected by RT-PCR, Western blot. The contents of nitrites and the levels of Th1 type cytokine IFN-γand Th2 type cytokine IL-4 were measured by Griess test and ELISA respectively, in the co-cultured supernatant. Results T-lymphocyte proliferation was inhibited by co-cultured MSCs, which was concentration-dependent. In this study, the inhibition was most obviously group[ (79.03 ± 1.70)% ] (P ＞ 0.05 ). The I NOS mRNA expression, protein and nitrite levels were signifigroups, the proliferation rate of T-lymphocyte recovered. The content of IFN-γwas increased with the ratio decline of MSCs in the experimental group and IL-4 in each group has no significant difference( P ＞ 0.05 ).Conclusion MSCs inhibited T-lymphocyte proliferation by influencing Th1/Th2 balance, and the secretion of soluble factor NO, which secreted by MSCs, may plays an important role in the immune regulation.

Objective To discuss the distributional characteristics and drug-resistance of Enterococcus species isolated from urine specimens.Methods 3096 middle-segment urine specimens were collected since January to December in 2011 for culture.The identification of pathogenic bacteria and antibiotics sensitivity tests were carried out with VITEK2-compact combined with GN,AST-GN13,GP,AST-GP67,and antibiotics sensitivity tested performed by K-B and E-test at the same time.The results were determined by CLSI 2011.Results 1248 of 3096 pathogenic bacteria were isolated (40.31％).549 strains of Escherichia coli were detected (43.99％) which was the most common and 159 strains of Enterococcus were detected (12.74％) which was the second most common.The Enterococcus detection rate in woman (15.02％)was higher than that in man (10.35％),in out-patients (15.54％) than the that in hospitalized patients (12.49％),and in the patients of non-surgical departments (13.65％) than those of surgical departments (11.38％).The Enterococcus was absolutely sensitive to tigecycline,and the sensitive rate to vancomycin and linezolid were over 90％.The antibiotics sensitivity was higher for Enterococcus faecalis than that for Enterococcus faecium,and in surgical departments than non-surgical departments.Conclusions The detection rate of Enterococcus in urinary tract infection patients is quite high and varied between sexes and departments.The difference of drug resistance between species is obvious,and the bacteria species should be identified in order to use the antibiotics reasonably to postpone the development of drug resistant strains.

ObjectiveTo evaluate the value of inferior petrosal sinus sampling (IPSS) in the diagnosis of adreno-corticotropic hormone ( ACTH ) dependent Cushing syndrome ( CS ) with unknown origin. MethodsIPSS was carried out for the diagnosis of 16 cases with ACTH dependent CS who had not been identified after a series of dexamethasone suppression tests and radiological examinations. The ratio of inferior petrosal sinus/peripheral ACTH was assayed.The sensitivity and specificity of diagnosis of the Cushing disease were estimated. ResultsThe inferior petrosal sinus/peripheral ACTH ratio was over 2. 0 in 13 cases. Twelve cases underwent surgery with pathological diagnosis of pituitary ACTH adenoma, 1 patient relieved after γ knife treatment. The ratio was ＜ 2.0 in 3 cases including 2 pulmonary carcinoid and one pituitary ACTH adenoma. The sensitivity and specify of IPSS for the diagnosis of Cushing disease were 13/14and 2/2 respectively.ConclusionIPSS was a safe technique with high sensitivity, specify and infrequent complications in the diagnosis of ACTH dependent Cushing disease.It had great clinical value in the differential diagnosis of ACTH dependent Cushing disease with unknown origin.

ObjectiveTo investigate the effect of bone marrow-derived mesenchymal stem cells (MSCs) on gene and protein expression of toll-like receptor, TLR2/4 of the rat alveolus macrophages so as to discuss the effect of MSCs transplantation on the early inflammatory mediators and the regulatory mechanism of the inflammatory cascade.MethodsThe alveolus macrophages were randomly divided into the PBS control group (n = 24), MSCs control group (n = 24), lipopolysaccharide (LPS) control group (n=24) and LPS + MSCs treatment group (n =24).All groups were taken time-points of 1, 3,6, 12 hours.Positive cell rate of MSCs with surface mark and the expression level of TLR2/4 protein on macrophage were detected by flow cytometry.The mRNA level of TLR2/4 expression on macrophage was detected by RT-PCR and the concentrations of TNF-o in cell culture supernatants by ELISA at the different time points.ResultsCompared with the PBS control group and MSCs control group, TLR2/4 mRNA and protein levels were significantly up-regulated, markedly increased at one hour and peaked at 12 hours (P ＜0.01) and the TNF-α concentrations in cell culture supernatants were significantly up-regulated and peaked at 6 hours in the LPS control group (P ＜ 0.01).Compared with the LPS control group, mRNA and protein level of TLR2/4 and TNF-o concentrations in the cell culture supernatants of the LPS + MSCs group were markedly down-regulated (P ＜ 0.01).ConclusionsWith LPS stimulation, TLR2/4 mRNA and protein expressions as well as TNF-αt concentration increase, when the peak of the TNF-αt concentration appears slightly earlier, indicating that there may be a positive feedback loop between TLR2/4 expression and TNF-ot.MSCs can significantly reduce TLR2/4 mRNA and protein expressions as well as TNF-α concentration after LPS stimulation, indicating that MSCs can break up the positive feedback loop.

ObjectiveTo observe the effects of electro-acupuncture combined neuromuscular electrical stimulation (NMES) on dysphagia in patients after acute cerebral infarction (ACI).Methods180 ACI patients with dysphagia were divided into the function training group (n=45)，electro-acupuncture group (n=45), NMES group (n=45) and comprehensive rehabilitation group (electro-acupuncture combined with NMES group, n=45). They were assessed with drinking water test before and 3 weeks after treatment. ResultsThe drinking water test scores improved after treatment in all the 4 groups (P<0.05), and more in the electro-acupuncture group, NMES group and comprehensive rehabilitation group than in the function training group (P<0.05). The drinking water test scores improved the most in comprehensive rehabilitation group (P<0.05). ConclusionThe electro-acupuncture combined NMES can effectively improve the therapeutic effect on dysphagia in patients after ACI.

OBJECTIVE:To study pharmacognosy of Melilotus officinalis.METHODS:Qualitative identification of M.officinalis was conducted in respects of morphology,macroscopic characters,microscopic identification,UV spectra for medicinal extracts and IR spectra for medicinal powder.RESULTS:Nineteen bundles of rays were distributed in the radial bundle of root cross section of M.officinalis.The main vein vascular bundle of leaf cross section had a palisade tissue passing over it.The pith of the cross section of the stem occupied 4/5 of the whole cross section.The cross section of the leafstalk was heart-shaped,and unequal vascular bundles arranged in a triangular array.There were glandular hairs,which consist of three cells,and unicellular non glandular hairs in leaf epidermis.The crystal fibers of calcium oxalate crystals and infinitive blowhole were found in the leaves;glandular hairs and non glandular hairs could be found in the leafstalk under tissue dissociation;verrucous like protuberance and threaded catheter were found on the surface.The UV spectrum of extract and two order derivative IR spectrum of the medicinal powder showed obvious characteristics.CONCLUSIONS:Established method can be used for pharmacognostic identification of M.officinalis.

Objective: To investigate the abnormalities of the blood system in landfill workers. Methods: A cohort study was conducted for 224 landfill workers who were followed up for 6 consecutive years with abnormal routine blood test results and a low platelet count as the outcome events. The life-table method was used to analyze the incidence rates of these two outcome events, and the incidence rates were compared between first-and second-line workers. Results: A total of 71 workers had abnormal routine blood test results, among whom 29 had abnormal leukocyte count, 14 had abnormal erythrocyte count, 40 had abnormal platelet count, 17 had abnormal hemoglobin, and 29 had a reduction in platelet count. For these landfill workers, the 6-year abnormal rate of routine blood test results was 43.2%, and the incidence rate of low platelet count within 6 years was 13.5%. The first-line workers had a significantly lower abnormal rate of routine blood test results than the second-line workers (P=0.020) , and the relative risk of abnormal routine blood test results in the first-and second-line workers was 0.592. There was no significant difference in the incidence rate of low platelet count between the two groups of workers (P≥0.05) . Conclusion The landfill has an adverse effect on the blood system of landfill workers, and the second-line workers have greater impairment than the first-line workers.

Objective: To explore the clinical application of the deep inferior epigastric artery perforator flap in bilateral breast reconstruction of patients with breast cancer. Methods: Deep inferior epigastric artery perforator flap was applied for breast reconstruction in four cases of breast cancer patients received radical or modified radical surgery in Hunan Provincial Cancer Hospital. All patients are female, age ranged from 31 to 53 years old (36.2±5.9). Results: The length of flap was (15.1±0.4) cm, the width of flap was (12.6±0.3) cm, the thickness of flap was (4.3±0.5) cm. The length of pedicle was (12.6±0.3) cm, the outer diameter of artery was (1.8±0.2) mm, the outer diameter of vein was (2.1±0.4) mm.The average weight of flaps was 235 g(ranged from 195 g to 335 g). In one case flap fat necrosis occurred and in other one donor site fat necrosis was noted. The two flaps both healed with dressing treatment and no other complications were found. The reconstructed breasts′shape, texture and elasticity were good and no flap contracture deformation happened. Only linear scar left in the donor sites, the function of abdomen did not affected. All 4 patients were followed up for 14 to 33 months (21.7 months on average) with satisfied result. No local recurrence happened. Conclusions Free deep inferior epigastric artery perforator flap is suitable to reconstruct bilateral breast for breast cancer patients.