Objective To study the effect of fusion protein FADDDEL-GFP in the treatment of type 1 diabetes by murine islet cell transplantion.Methods After transfecting the recombinate FADDDEL-GFP into mouse insulinoma cells NIT-1,insulin level secreted by the cells was examined.Type 1 diabetes was induced by STZ as animal model.NIT-fg cells,the NIT-1 cells modified by FADDDEL-GFP,were transplanted into the diabetic mice.Then the effects of islet cell transplantation on diabetic mice were checked.Results GFP was expressed in NIT-1 cells transfected with FADDDEL-GFP.Diabetic mice reached normoglycemia and prolonged the life span,which was obviously different from the control group.Conclusion FADDDEL-GFP may enhance the capability of resisting allogeneic transplantation rejection.

Objective To investigate awareness of hypertension prevention and treatment knowledge in physicians from hospitals at varied levels in Xinjiang. Methods In total, 150 voluntary physicians were selected randomly from hospitals at varied levels in Xinjiang for an anonymous close-book baseline survey on hypertension knowledge with questionnaire. Then, an intensive training on hypertension prevention and treatment based on Guidelines on Prevention and Treatment for Hypertension in China was offered for them.After training, another survey was conducted among them with the same questionnaire to examine improvement in their awareness of hypertension prevention and treatment knowledge and evaluate effectiveness of the training. Results At baseline, 89. 3 percent (134/150) of physicians could correctly know diagnostic criteria for hypertension, 78. 3 percent ( 18/23 ) of them from primary-care hospitals, and 52. 0 percent (78/150) could correctly know level of blood pressure under control, only 34. 8 percent (8/23) of them from primary-care hospitals. Only 67 (44. 7% ) physicians surveyed could know criteria for non-antihypertensive drug treatment, 27 of then from secondary-care hospitals and nine from primary-care ones, significant less in that among those from tertiary-care ones ( 88. 6%, 31/35 ) ( P ＜ 0. 05 ). After training, their awareness of hypertension prevention and treatment knowledge improved significantly ( P ＜0. 01). Conclusions Awareness of hypertension prevention and treatment knowledge differed considerably in physicians from hospitals at varied levels, poorer in those from primary-care hospitals, and more importance should be attached to them, especially to those from primary-care hospitals.

Aim To investigate the role of GlyR-α3 in postoperative pain.Methods The model of postoper-ative pain was established by incision of mouse skin and muscle.Pain responses including paw withdrawal thresholds(PWTs),paw withdrawal latencies(PWLs)and licking toes time were conducted and recorded.Protein contents and tyrosine phosphorylation levels of GlyR-α3 were detected by co-immunoprecipitation and immunoblotting.Bosutinib was used to inhibit Src ki-nase activity to verify whether Src was involved in tyro-sine phosphorylation of GlyR-α3.Results The PWTs and PWLs in postoperative pain model mice were sig-nificantly reduced,whereas the licking time increased significantly.The tyrosine phosphorylation level of GlyR-α3 in the dorsal root ganglia(DRG)from inci-sion sidesignificantly increased compared with contra-lateral side,while the protein expression showed no sta-tistical significance.The expression of Src in DRGin-creased after operation,and the interaction between Src and GlyR-α3 was enhanced in ipsilateral DRG com-pared with contralateral side.Inhibition of Src kinase activity helped to reduce tyrosine phosphorylation of GlyR-α3 from DRG and alleviate postoperative pain symptoms.Conclusion Src kinase is involved in the development of postoperative pain by enhancing tyro-sine phosphorylation of GlyR-α3.

BACKGROUND: The virulent factors ofEscherichia coli (E.coli) play an important role in the process of pathopoiesis. The study aimed to compare drug-resistant genes and virulence genes between extended spectrum β-lactamases (ESBLs)-producingE.coli and non-ESBLs-producing E.coli to provide a reference for physicians in management of hospital infection. METHODS: From October 2010 to August 2011, 96 drug-resistant strains ofE.coli isolated were colected from the specimens in Qingdao Municipal Hospital, Qingdao, China. These bacteria strains were divided into a ESBLs-producing group and a non-ESBLs-producing group. Drug sensitivity tests were performed using the Kirby-Bauer (K-B) method. Disinfectant gene, qacEΔ1-sull and 8 virulence genes (CNF2, hlyA, eaeA, VT1, est, bfpA, elt, and CNF1) were tested by polymerase chain reaction (PCR). RESULTS: Among the 96E.coli isolates, the ESBLs-producingE.coli comprised 46 (47.9％) strains and the non-ESBLs-producingE.coli consisted of 50 (52.1％) strains. The detection rates of multiple drug-resistant strain, qacEΔ1-sull, CNF2, hlyA, eaeA,VT1, est, bfpA, elt, and CNF1 in 46 ESBLs-producingE.coli isolates were 89.1％, 76.1％, 6.5％, 69.6％, 69.6％, 89.1％, 10.9％, 26.1％, 8.7％, and 19.6％, respectively. In the non-ESBLs-producingE.coli strains, the positive rates of multiple drug-resistant strain, qacEΔ1-sull, CNF2, hlyA, eaeA, VT1, est, bfpA, elt, and CNF1 were 62.0％, 80.0％, 16.0％, 28.0％, 64.0％, 38.0％, 6.0％, 34.0％, 10.0％, and 24.0％, respectively. The difference in the detection rates of multiple drug-resistant strain, hlyA and VT1 between the ESBLs-producingE.coli strains and the non-ESBLs-producingE.coli strains was statistically significant (P<0.05). CONCLUSION: The positive rate of multiple drug-resistant strains is higher in the ESBLs-producing strains than in the non-ESBLs-producing strains. The expression of some virulence genes hlyA and VT1 varies between the ESBLs-producing strains and the non-ESBLs-producing strains. Increased awareness of clinicians and enhanced testing by laboratories are required to reduce treatment failures and prevent the spread of multiple drug-resistant strains.

Objective: To summarize the influence patterns of related time factors on acupuncture effectiveness in different disease model rats/mice, and to provide reference for acupuncture clinical practice. Methods: Retrieved the relevant literatures on time-effect experimental studies of acupuncture in rats/mice in the recent 10 years. The correlations between the key time factors (such as different intervention timings of acupuncture, acupuncture moments, operation durations, needle-retaining times, intervals and treatment courses) and the acupuncture effect were analyzed and summarized. Results: From the mainstream perspective of quantification, the earlier the acupuncture intervention, the better. The proper time to implement acupuncture varied depending on disease models and points. The best operation time varied widely between different diseases. The most frequently needle-retaining time was 20-30 min. The frequency of acupuncture was usually 1 time/day; the length of the treatment course was determined according to practitioners' experience. Conclusion: Throughout the time-effect studies of acupuncture intervention in experimental rats/mice, conclusions are inconsistent, especially the lack of quantitative research on acupuncture operation time, acupuncture frequency, acupuncture treatment duration, and optimal stimulation amount. Future research should explore and determine the best time-quantity parameters that affect the effectiveness of acupuncture intervention, which is the key and goal of the acupuncture time-effect research. Independent intervention-time experiments throughout the entire course of a single disease (dominant disease) need to be done to guide clinical and disciplinary development.

Objective To investigate the expression of Foxp3~+ lymphocytes in breast carcinoma tissues and their correlation with other pathological factors,and to investigate the mechanism of action of Treg cells.Methods The expression of Foxp3~+ lymphocytes in the breast cancer tissue and non-cancerous tissue was detected by flow cytometry (FCM) in 30 breast carcinoma patients, and its correlation with other pathological factors was statistically analyzed by multiple linear regression analysis.The expression of TGF-β and IL-10 in the lymphocytes infiltrated in breast cancer tissue and non-cancerous tissue was measured by immunohistochemistry, and their correlation with the expression of Foxp3~+ lymphocytes was statistically analyzed by linear correlation dependability analysis. Results There was significant difference in the expression of Foxp3~+ lymphocytes between the malignant and non-cancerous breast tissues(P<0.05),and it was positively correlated with the clinical stage,blood vessel invasion and the matter of axillary lymph node metastasis(P<0.05). The expression of IL-10 in the tumor infiltrating lymphocytes was positively correlated with the expression of Foxp3~+ lymphocytes(P<0.05).Conclusion The expression level of Foxp3~+ lymphocytes is correlated with invasion and metastasis of breast carcinoma, and the IL-10 secreted by Foxp3~+ lymphocytes may be involved in this effect.Foxp3~+ lymphocytes can be used as an assistant marker for prediction and new therpeutic target of breast cancer.

Objective To analysis the quality of life of patients with active ankylosing spondylitis (AS).Methods The quality of life was assessed in 52 patients with active AS using SF-36 and was compared with the general population.The correlation between the quality of life and the clinical measures of disease,in- cluding the Bath AS disease activity index(BASDAI),Bath AS functional index(BASFI),Bath AS metrology index(BASMI),patient's global assessment(PGA),spinal inflammation,total back pain,nocturnal back pain, and enthesis index(EI),were determined.Results The patients with active AS reported significant decreased scores on all dimensions of SF-36.The score of physical health components was poorer than that of mental health components.BASFI was the strongest factor correlated with the score of SF-36,followed by BASDAI and PGA.BASMI and total back pain were correlated with three dimensions only.In multivariate regression analysis,BASFI showed relative closer relationship to the qulity of life with active AS than other clinical mea- sures of disease and it accounted for 50.3%,35.2% and 47.9% of the variance in the physical health compo- nents,the mental health components and the overall score of SF-36,respectively.Conclusion The quality of life in patients with active AS is significantly declined compared with general population.The physical aspects seem to be more severely affected.Functional status of the patients with active AS are correlated with the quality of life closely.

Objective To investigate immunophenotypic characteristics of uterine natural killer (uNK)cells and helper T cell 1/helper T cell 2(Th1/Th2)immunity in third trimester decidua in preeclampsia.Methods The proportions of uNK cell subsets,expression of CD_(69)and CD_(94)on uNK cells and Th1/Th2 immunity in decidua were determined in 20 cases of preeclampsia patients and 11 cases of normal term pregnancies by flow cytometric analysis.Results The percentage of CD_(56)~(bright)CD_(16)~-uNK cell subset in preeclampsia patients and the controls was(17.3?11.1)% vs(17.9?16.8)%,that of CD_(56)~(dim) CD_(16)~+uNK cell subset was(16.3?8.7)% vs(16.2?8.8)%;that of CD_(56)~+CD_(69)~+uNK cells was(37.9 ?18.9)% vs(36.8?19.7)%,that of CD_(56)~+CD_(94)~+uNK cells was(34.9?15.2)% vs(32.7?16.2)% and the ratio of CD_(56)~+CD_(69)~+/CD_(56)~+CD_(94)~+was 1.1?0.2,1.2?0.6.No statistical difference was shown in the above values between the preeclampsia patients and controls.The percentage of cytoto xic T cell(Tc)2 cells was significantly lower in the decidua of preeclampsia patients [(3.0?1.0)% vs(4.3?0.9)%,P= 0.001 ],and the ratio of Tc1/Tc2 in preeclampsia patients was significantly higher than that of normal term pregnancies(17.8?3.4 vs 11.8?4.6;P=0.001);the ratio of Th1/Th2 was increased(15.1?2.4 vs 13.2?3.1;P=0.06).Conclusions The immunophenotypie characteristics of uNK cells do not present any significant change in preeclampsia patients.Owing to Tc2 cell decrease,the Th1/Th2 immunity shifts to Th1 type immunity in the decidua,which might contribute to the pathogenesis of preeclampsia.

The expression of N-myc down-regulated gene 1 (NDRG1) has previously been reported to be involved in the proliferation, differentiation, invasion and metastasis of cancer cells, but its role in cervical cancer is still unclear. This study aimed to investigate the expression of NDRG1gene in human cervical cancer and its effect on aggressive tumor behaviors. The NDRG1 expression in cervical tissues and cells was detected by RT-PCR. Specific expression plasmid pEGFP-N1-NDRG1-GFP was used to enhance the expression of NDRG1 in human cervical cancer cell lines. The mRNA and protein level of NDRG1 was assessed by RT-PCR and Western blotting, respectively. Its effects on cell proliferation, migration, invasion, cell cycle and apoptosis were detected by MTT, transwell migration assay and flow cytometry (FCM), respectively. The results showed that the expression of NDRG1 in cervical cancer tissues and cells was significantly lower than in normal cervical tissues (P<0.001). After transfection with pEGFP-N1-NDRG1-GFP, the mRNA and protein expression of NDRG1 was up-regulated in Siha cells, which suppressed cell proliferation (P<0.001), induced cell cycle arrest (P<0.05), reduced invasion and migration of Siha cells (P<0.05), but caused no cell apoptosis. Moreover, vascular endothelial growth factor (VEGF), a tumor-induced angiogenesis factor, was markedly reduced and E-cadherin, a cell adhesion molecule, was increased in the cells transfected with pEGFP-N1-NDRG1-GFP. It was concluded that up-regulated NDRG1 may play a role in the suppression of malignant cell growth, invasion and metastasis of human cervical cancer.

Objective To evaluate the role of complement receptor 3 (CR3) on murine macrophages in the recognition of Penicillium marneffei.Methods RAW264.7 murine macrophage cells were cultured in vitro,and divided into four groups to be cocultured with inactivated and live Penicillium mameffei yeast cells as well as inactivated and live Penicillium marneffei conidia respectively at 37 ℃ in 5％ CO2 for one hour.The RAW264.7 cells incubated with phosphate-buffered saline (PBS) served as the blank control group.Then,reverse transcription-PCR was conducted to detect CR3 mRNA expression,Western blot to measure CR3 protein expression,flow cytometry to determine phagocytosis rate,enzyme-linked immunosorbent assay (ELISA) to quantify cytokine levels in culture supernatant.Some RAW264.7 macrophages were transfected with a specific siRNA targeting CR3 gene and cocultured with inactivated Penicillium marneffei conidia,subsequently,phagocytosis rate and supematant cytokine levels were determined.Data were processed by the SPSS 16.0 software,and one-way analysis of variance (ANOVA) was conducted for inter-group comparisons of these parameters.Results No significant differences were observed in the mRNA or protein expressions of CR3 among the four groups of RAW264.7 cells cocuhured with different forms of Penicillium marneffei (both P ＞ 0.05).The phagocytosis rate was 95.14％,89.56％,91.03％ and 90.78％ in RAW264.7 cells cocultured with inactivated conidia and yeast cells,as well as live conidia and yeast cells of Penicillium marneffei,respectively (P ＞ 0.05).The levels of interleukin (IL)-2,interferon (IFN)-γ,IL-4 and IL-10 in culture supernatant were increased at different degrees after one-hour coculture in the four coculture groups compared with the blank control group,but no statistical difference was noted among the four coculture groups in the supernatant levels of these cytokines (all P ＞ 0.05).After coculture with inactivated Penicillium marneffei conidia,the siRNA-transfected RAW264.7 cells showed a statistical decrease in phagocytosis rate (10.89％ vs.92.78％,P ＜ 0.05) and supernatant levels of IL-2,IFN-γ IL-4 and IL-10 compared with untransfected RAW264.7 cells.Conclusions In early stage of innate immunity,CR3 on macrophages may be one of the pattern recognition receptors participating in the recognition and mediation of phagocytosis of Penicillium marneffei.It's possible that both Thl-and Th2-type cytokines,such as IL-2,IFN-γ,IL-4 and IL-10,are involved in the immune response of macrophages against Penicillium marneffei.