Objective To estimate the degree of oxidative stress and atherosclerosis praecox in patients with systemic lupus erythematosus (SLE), and emphasize on exploring AOPPs' clinical significance in premature atherosclerosis in SLE. Methods The levels of AOPPs, Hey, MDA, SOD, baPMV and CIMT were detected by ELISA and spectropholometry in 44 non-menopausal female SLE patients and 31 healthy middle-aged women respectively, and baPWV. The results were compared with AOPPs of the two groups. Then each group was stratified based on disease duration (≥5 years or <5 years) and the disease activity(active and inactive) in SLE patients. The patients' TC, TG, LDL were analyzed. T test, t' test and Pearson correla-tion were selected. Results The levels of AOPPs, Hcy, MDA in SLE patients were significantly higher than those of the healthy controls (P<0.05). The activities of SOD were lower than controls (P<0.05). The levels of AOPPs, Hcy, MDA, SOD had statistical significance between SLE patients disease duration ≥5 years or <5 years, active or inactive groups. There were two cases with CAP in patients with SLE (more than 5 years disease duration),while there wasnone in healthy controls. The levels of baPWV and CIMT in patients with SLE were higher than healthy controls (P<0.05), and had statistical significance in SLE patients disease duration more than or less than 5 years (P<0.05). The oxidative stress targets (AOPPs, Hey, MDA, SOD) had significant correlation with the level of baPWV and CIMT (P<0.01). The level of serum AOPPs had significant positive correlation with the levels of Hcy, TC, TG and LDL (P<0.01～0.05). Conclusion SLE patients have increased oxidative stress , and significantly higher prevalence of atherosclerosis than healthy controls. The disease duration and oxidative stress play important roles in the duration of atherosclerosis. AOPPs probably involves in the accelerated atherosclerosis of SLE patients. It may be a predictor for SLE complicated with atherosclerosis praecox.

Objective To explore the clinic value and efficacy of color Doppler ultrasound guided percutaneous microwave/radiorequency thermoablation for secondary hyperparathyroidism. Methods Fifty-six cases of secondary hyperparathyroidism who came from nephrology department of Beijing Friendship Hospital in the period of September 2012 to September 2013 were treated by color Doppler ultrasound guided percutaneous microwave/radiorequency thermoablation. One hundred and thirty-eight glands underwent this treatment and were evaluated by contrast enhanced ultrasound before, during and after operation. The levels of serum parathyroid hormone (PTH) were measured before and at 1 week, 1 month, 6 months and 1 year after treatment. Results The whole 56 cases achieved 1 month follow-up and 34 cases and 12 cases achieved 6 months and 1 year follow-up, respectively. Two special cases had been excluded. The PTH levels of 54 cases were signiifcantly different between after and before treatment [(369.5±183.4) ng/L vs (1321.6±471.4) ng/L, t=10.727, P=0.000]. The PTH levels continued to fall at the ifrst week after treatment, but there was no statistical signiifcance between the ifrst week and the ifrst day after treatment [(324.6±172.8) ng/L vs (369.5±183.4) ng/L], while there was signiifcantly different between the ifrst week after treatment and before treatment [(324.6±172.8) ng/L vs (1321.6±471.4) ng/L, t=9.364, P=0.000]. The PTH levels increased from (324.6±172.8) ng/L to (332.5±164.9) ng/L at 1 week and 1 month after ablation, while there was no signiifcant difference.But there is signiifcantly different between 1 month after treatment and before treatment (t=11.348, P=0.000). The PTH level at 6 months after treatment [(205.2±136.3) ng/L] and 1 year after treatment [(177.1±171.5) ng/L] had signiifcant difference compared with the value before treatment (t=8.737, P=0.000; t=7.655, P=0.017). Conclusion Color Doppler ultrasound guided percutaneous microwave/radiorequency thermoablation can be considered as a feasible and effective nonsurgical alternative treatment for secondary hyperparathyroidism patients.

Endostatin is a potent, endogenous inhibitor of angiogenesis, which corresponds to the C-terminal fragment of collagen ⅩⅧ. The human endostatin gene was amplified from a human fetal liver cDNA library by means of PCR and was cloned into pPIC9K vector. Soluble endostatin was superexpressed in Pichia pastoris (50.5mg/L) . The recombinant protein was purified by cation exchange chromatography with the final yield of more than 95%. Western blotting analysis showed positive immunoreactivity to the expressed product. Recombinant endostatin was able to suppress the angiogenesis on the CAMs. Also, the endostatin inhibited specifically of the migration of HMECs stimulated by bFGF, with EC50 being about 0.4 ?g/ml.

Objective To explore the expression,correlation and clinical significance of mammalian target of rapamycin (mTOR) and vascular endothelial growth factor (VEGF) in gastroenteropancreatic neuroendocrine neoplasm (GEP-NEN).Methods From October 1995 to December 2012,the paraffin tissue specimens of 114 pathological diagnosed GEP-NEN were collected.The expressions of mTOR and VEGF at protein level were detected with EnVision immunohistochemical staining method.Chi-square test was performed to compare the differences in the expressions of mTOR and VEGF among every clinicopathological factors of patients.Spearman's correlation analysis was used to find out the correlation between the expressions of mTOR and VEGF.Kaplan Meier method and Log-rank test were implemented for survival analysis.Results The overall positive expression rates of mTOR and VEGF in GEP-NEN tissues were 66.7％ (76/114) and 71.1％ (81/114),respectively.The expressions ofmTORand VEGF at protein level were not correlated with gender,age,functional status of tumor,location of tumor,grading and classification of tumor and distant metastasis (all P＞0.05).The co-expression rate of mTOR and VEGF in GEP-NEN tissues was 51.8％ (59/114).The expression of mTOR was positively correlated with the expression of VEGF at protein level (r=0.205,P=0.029).Among 82 followed-up patients,the mean survival time of patients with mTOR positive or negative expression was 8.3 and 7.9 years,and respectively,and there was no statistical significance (x2 =0.201,P=0.654).The mean survival time of patients with VEGF positive or negative expression was 9.6 and 5.9 years,respectively,and the difference was statistically significant (x2 =4.415,P=0.036).Among the patients with G1 grading tumor,classified as neuroendocrine tumor and without distant metastasis,the survival time of patients with positive VEGF expression was longer than that of patients with negative VEGF expression (11.8 years vs 7.3 years,x2 =5.349,P=0.021; 11.2 years vs 6.8 years,x2=6.375,P=0.012; 11.2 years vs 7.2 years,x2 =4.630,P=0.031).Conclusions mTOR and VEGF proteins are highly expressed in GEP-NEN with different location,grading and classification.The expression of them is positively correlated.The prognosis of GEP-NEN patients with positive VEGF expression is better.

Objective To explore the occurrence site, clinical manifestations, endoscopic and radiographicfeatures, pathologiccharacteristics, diagnosis, treatmentandprognosisof gastroenteropancreatic neuroendocrine neoplasm (GEP-NEN).MethodsFrom January 2000 to June 2010,medical records of 87 GEP-NEN patients in the First Affiliated Hospital of Sun Yat-sen University were collected. The neuroendocrine properties of the tumors were determined by immunohistochemistry staining of neuroendocrine markers Syn and CgA.Then according to histology and proliferation activity, the GEP-NEN tumors were classified as neuroendocrine tumor (G1 and G2 grade), neuroendocrine carcinoma (G3 grade) and mixed adenoendocrine carcinoma (G3 grade).Results36 cases (41.4％) of GEP-NEN tumors were in pancreas, followed by 18 cases in rectum (20.7％), 9 cases in stomach (10.3％) and 6 cases in duodenum (6.9％).Total 69 of 87 GEP-NEN cases (74.7％) were nonfunctional tumors, a variety of gastrointestinal symptoms or symptoms caused by local tumor compression were the most first symptoms, and no one case presented carcinoid syndrome.The manifestations of endoscopy and radiographic image were tumor occupying lesion.Of 87 GEP-NEN cases, 69 cases were neuroendocrine tumor (79.7％), 13 cases were neuroendocrine carcinoma (14.9％) and 5 cases were mixed adenoendocrine carcinoma (5.4％).The percentage of grade G1, G2 and G3 tumors were 64.9％、 14.9％ and 20.2％ respectively.Muscularis or serosa infiltration were found in 22 GEP-NEN patients (47.8％) at the first visit, lymphatic metastasis was in 18 cases (20.7％) and distant metastasis were in 18 cases (20.7％), mostly migrated to liver.Immunohistochemistry staining positive rates of CgA and Syn were 74.2％ and 88.1％.A total of 79 patients (90.8％) were underwent surgery.The 1-, 3-and 5-year survival rates were 76.9％, 54.2％and 41.7％ during follow-up.ConclusionsGEP-NEN may occur at any part of the digestive system,the clinical manifestations are variable.Endoscopy and radiographic examination play an important role in tumor diagnosis, but final diagnosis is mainly based on pathological detection, and surgery is the main treatment option.The prognosis of the tumor is correlated with the pathological classification,grading and condition of distant metastasis.

Objective To explore the value of serum chromogranin A (CgA) in clinical diagnosis and efficacy evaluation of gastroenteropancreatic neuroendocrine neoplasm (GEP-NEN).Methods From January 2011 to December 2012,87 GEP-NEN patients were enrolled,which included 50 patients with lesions and 37 patients without lesions after surgery.Eighty-four healthy subjects were taken as control group.The serum CgA levels of all subjects were measured with enzyme linked immunosorbent assay.Nonparametric test,receiver operating characteristic curve and Cox regression univariate analysis were performed for analysis.Results The median of serum CgA level of GEP-NEN patients with lesions was 97μg/L,which was significantly higher than that of patients without lesions after surgery (42 μg/L,Z=3.451,P=0.001) and healthy control (47 μg/L,Z=3.149,P=0.002).The value of 95 μg/L was taken as diagnostic cut-off value for healthy controls,patients without lesions and patients with lesions,the sensitivity and specificity were 54.0％ and 90.1％,respectively.The median of serum CgA level of the patients with distant metastasis was 231 μg/L,which was higher than that of patients without distant metastasis (46 μg/L,Z=3.340,P=0.001).After treatment,the serum CgA levels of 12 patients with complete remission,partial remission or stable tumor decreased more than 20％ of baseline values.The serum CgA levels of five patients increased more than 20 ％ of baseline values and the tumors of them showed progress.Cox regression analysis showed that there was no correlation between CgA levels of patients with lesions and prognosis (r=1.000,P=0.252).Conclusion Neuroendocrine marker CgA has high diagnostic value in GEP-NEN,and can be used as an important biomarker in clinical diagnosis and efficacy evaluation of GEP-NEN.

Objective To investigate the effects and mechanisms of 1,25-Dihydroxyvitamin D3 [1,25(OH)2 D3] on hyperoxia-induced lung injury of neonatal rats.Methods Neonatal Sprague-Dawley rats were randomly divided into air group,hyperoxia group and 1,25(OH)2D3 group within 12 hours after birth,eight in each group.Rats in air group were exposed to air,while those in hyperoxia and 1,25 (OH) 2 D3 group were exposed to hyperoxia (≥85 ％ oxygen concentration).Rats in 1,25(OH)2D3 group were injected with 1,25(OH)2D3 0.5 μg/(kg · d) intraperitoneally once a dayfor seven days,meanwhile the rats in the other two groups received 0.9 ％ saline in the same way.All rats were sacrificed on day 7.Lung tissue sections were HE stained in order to assess lung histological changes and lung radical alveolar count (RAC).The levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-1β and IL-6 mRNA were measured by real time-polymerase chain reaction.Analysis of variance and LSD test were applied for statistics.Results Compared with the air group,the weight of rats in the hyperoxia group was significantly lower on day 7 [(8.48±1.34) g vs (12.51±0.47) g,t=8.05,P＜0.05],while the weight of rats in 1,25(OH)2D3 group [(10.29±1.00) g] was higher than that in the hyperoxia group (t=3.61,P＜0.05).Lung tissue structure was normal in the air group.In the hyperoxia group,inflammatory exudation was observed in pulmonary interstitial,the alveolar size was uneven,and the RAC was lower than that in the air group (5.6±0.1 vs 6.8±0.2,t=21.45,P＜0.05).The RAC in 1,25(OH)2D3 group (6.2±0.1) was significantly increased compared with that in the hyperoxia group (t=11.76,P＜0.05),but still lower than that in the air group (t=9.69,P＜0.05).The expressions of TNF-α,IL-1β and IL-6 mRNA in hyperoxia group (0.0348±0.0006,0.0269±0.0003 and 0.0368 ± 0.0006) were higher than those in the air group (0.0111±0.0007,0.0040±0.0003 and 0.0162 ±0.0007,t=56.54,111.12 and 49.26,P＜0.05,respectively).The expressions of TNF-α,IL-1β and IL-6 mRNA in the 1,25 (OH)2D3 group (0.0203±0.0009,0.0141±0.0004 and 0.0251±0.0009) were lower than those in the hyperoxia group (t=34.44,61.93 and 27.99,P＜0.05,respectively),but higher than those in the air group (t=22.10,49.19 and 21.27,P＜0.05,respectively).Conclusions 1,25(OH)2D3 could attenuate hyperoxia-induced lung injury by inhibiting the expression of inflammatory cytokines.

[Objective] To investigate the expression of NF-κB and IL-lra in gastric cancer, and to explore their correlation with the clinicopathological parameters and prognosis of gastric cancer (GC). [Methods] Expression of NF-κB and IL-1ra was detected by immunohistochemistry in tissue microarry of 359 cases of GC. [Results] The expression rates of NF-κB in the patients with metastasis of lymph node, TNM Ⅲ-Ⅳ stage, and poorly differentiated of histology were 80.2%, 80.0%, and 79.2%, respectively. The expression rates of IL-1ra in the patients with tumor size ≤3 cm, early stage, non-metastasis of lymph node, and TNM Ⅰ-Ⅱ stage were 61.7%, 75.0%, 66.4%, and 61.9%, respectively. The 5-year survival rate of the cases with non-expressed NF-κB was statistically higher than that of the cases with expressed NF-κB(P=0.036). The 5-year survival rate of the patients with negative expression of NF-κB and positive expression of IL-1ra was statistically higher than the others (P=0.021). [Conclusions] NF-κB is the adverse predictors of prognosis of gastric cancer. IL-1ra maybe play a protective role in early gastric cancer stage, but it is necessary to study deeply and get more evidences.

To prepare virus-like particles containing FluA/B mRNA as RNA standard and control in Influenza RNA detection, the genes coding the coat protein and maturase of E. coli bacteriophage MS2 were amplified and cloned into D-pET32a vector. Then we inserted 6 histidines to MS2 coat protein by QuikChange Site-Directed Mutagenesis Kit to construct the universal expressing vector D-pET32a-CP-His. In addition, the partial gene fragments of FluA and FluB were cloned to the down-stream of expressing vector. The recombinant plasmid D-pET32a-CP-His-FluA/B was transformed to BL21 with induction by IPTG. The virus-like particles were purified by Ni+ chromatography. The virus-like particles can be detected by RT-PCR, but not PCR. They can be conserved stably for at least 3 months at both 4 degrees C and -20 degrees C. His-tagged virus-like particles are more stable and easier to purification. It can be used as RNA standard and control in Influenza virus RNA detection.
Escherichia coli ; genetics ; metabolism ; Influenza A virus ; genetics ; metabolism ; Influenza B virus ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; RNA, Viral ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Ribonucleases ; chemistry ; Virion ; genetics ; metabolism

Purpose To study the malignant transformation after treating rat oval cell line ( WB-F344 ) with chemical carcinogen N-methyl-N′-nitro-N-nitrosoguanidine ( MNNG) . Methods WB-F344 cells were cultured with MNNG for severe times. The biological characteristics of induced cells were detected through the following methods:to check proliferation activity by flow cytometry analysis, to examine malignant transformation degree of induced cells by soft agar assay and tumor formation in NOD/SCID mice, and to investi-gate the transcriptional and protein levels of hepatocellular carcinoma marker GGT, GST-P by real time-PCR. Results Oval cells in-duced by MNNG showed changes in biological characteristics and malignant molecular markers. Conclusion Hepatic oval cells model is successfully established, which can be confirmed by tumor formation in NOD/SCID mice.