Objective To observe the occurrence of vascular calcification in maintenance hemedialysis (MHD) patients. Method Sixty-nine MHD patients were enrolled in this study. Vascular calcification was evaluated by plain X-my films of chest, upper and lower extremites. Analyzed the relationship of vascular calcification and cardiovascular disease. Results Vascular calcification on X-ray films presented in 38 cases (55%). Of the 23 cases with calcification on arteries of extremites, 14 cases were found to have both intimal calcification and medial calcification. Patients with vascular calcification were older. Ischemic heart disease and cardiac failure appeared more frequently in the patients with vascular calcification. Conclusions Both intimal calcification and medial calcification are frequently found in the MHD patients. Vascular calcification is probablly associated with cardiovascular disease in MHD patients.

Objective To explore the possible correlations under the interaction of multi-genes action at different stages and analyze the primary histophotomacrographic changes of hind buds tissue in congenital clubfoot and the pathodynamic developmental procedure.MethodsSeventy-seven female Wistar rats were administered with retinoic acid on the 10th day after pregnancy. And the hindlimb, buds and spinal cord were detected through transmission electron microscopic and molecular biological experiments.ResultsThere was clubfoot-like deformity in 61.8% of the experimental animals. Persistence of the embryonic position of the talus and tibia in fetuses was observed. Poor overlapping between talus and calcaneus was seen. Cell apoptosis at the anterior corner of spinal cord and hind buds were seen.ConclusionCongenital clubfoot deformities appear at early stage and exaggerate along with developmental procedure.

Objective To explore the effect of three-dimensional conformal radiation dose fractionation treatment of advanced non-small cell lung cancer(NSCLC).Methods 75 NSCLC patients were treated with hypofractionated 3DCRT( observation group),while 73 cases were treated with conventional fractionated radiotherapy (control group).The efficacy and adverse reactions were observed;Survival after treatment were followed up in 1,2,3 years.Lung function was detected before and after radiotherapy treatment,including FVC,FEV1 and CLCO.Results The total effective cases of control group were 47 cases patients ( 64.4％ ),observation group's was 60 cases ( 80.0％ ),total effective rate had statistically significant difference ( x2 =4.50,P ＜ 0.05 ).Survival of control group after treatment in 1,2,3year were 50.7％,24.7％,8.2％,the median survival was 13 months,observation group's were 73.3％,45.3％,20.0％,and 19 months.The survival of these two groups was statistically different (x2 =8.07,6,94,4.22,all P ＜ 0.05 ).The patients blood system side effects of observation group were significantly lower than the control group ( x2 =4.73,P ＜0.05 ) ;acute radiation pneumonia,esophagitis of these two groups had no significant difference in the incidence( P ＞ 0.05 ).Conclusion Hypofractionated 3DCRT treatment of advanced NSCLC had good effect,and its adverse reactions was low,and it was worthy of clinical application.

Objective To investigate the relation between serum adiponectin and coronary artery calcification score (CACS), and find the risk factors for CACS in patients with chronic kidney disease (CKD). Methods Twenty-nine patients with 3-5 stage CKD were selected. The serum adiponectin was measured by enzyme linked immunosorbent assay. The heart was scanned by 64-row spiral CT, and the CACS was calculated. The blood calcium, phosphorus, intact parathyroid hormone, total cholesterol, low density lipoprotein cholesterol, albumin, urea nitrogen, creatinine, uric acid and high sensitive C reactive protein levels were measured, and the calcium-phosphorus product and estimation glomerular filtration rate were calculated. Results In 29 patients with CKD, 24 cases (83%) had coronary artery calcification with different degree (CACS>0 score), and the average CACS was 508 (0-3 363) scores. There were statistical differences in systolic blood pressure, urea nitrogen and estimation glomerular filtration rate between CKD patients with CACS≥100 scores (15 cases) and CKD patients with CACS<100 scores (14 cases):(146.00± 13.00) mmHg (1 mmHg=0.133 kPa) vs. (123.00±9.00) mmHg, (15.44±8.36) mmol/L vs. (9.71±2.52) mmol/L and (21.77 ±11.81) ml/ (min·1.73 m2) vs. (38.71 ±11.56) ml/ (min·1.73 m2), P<0.01 or <0.05. Pearson correlation analysis results showed that CACS had positive correlation with systolic blood pressure, creatinine and uric acid, and negative correlation with albumin and estimation glomerular filtration rate. Mult-stepwise regression analysis results showed that systolic blood pressure and estimation glomerular filtration rate were the independent risk factors of CACS. Conclusions The patients with 3-5 stage CKD have severe coronary artery calcification. The systolic blood pressure and estimation glomerular filtration rate are the independent risk factors of coronary artery calcification.

Objective To investigate the clinical significance of body composition monitor (BCM) in evaluating the dry weight of hemodialysis patients.Methods Fifty-four hemodialysis patients who reached the dry weight according to clinical assessment were enrolled.The volume load was measured with BCM,and the pre-dialysis blood pressure,plasma B type natriuretic peptide (BNP) and echocardiogram parameters were recorded.Results BCM results showed that,in 54 hemodialysis patients,15 patients presented overhydration and 13 patients underhydration.According to the results of BCM,all of the 15 overhydration patients and some of the 13 underhydration patients adjusted their dry weight.According to the BCM results and the pre-dialysis blood pressure,the patients were divided into 4 groups:group A (without overhydration,the pre-dialysis blood pressure ＜ 150 mmHg,1 mmHg =0.133 kPa),23 cases;group B (without overhydration,pre-dialysis blood pressure ≥ 150 mmHg),16 cases;group C (overhydration,pre-dialysis blood pressure ＜ 150 mmHg),7 cases;and group D (overhydration,pre-dialysis blood pressure ≥ 150 mmHg),8 cases.There were no significant differences in pre-dialysis,plasma BNP and the echocardiographic parameters among the 4 groups (P ＞ 0.05).Conclusions BCM is a simple and convenient tool to evaluate the volume status of hemodialysis patients,It can give guidance of dry weight regulation.Compared with the other clinical assessment methods such as pedal edema,the blood pressure and plasma BNP,it is more accurate and practical.

Objective To investigate the role of serum and glucocorticoid regulated protein kinase (SGK) 1 in the inflammatory responses mediated by toll like receptors.Methods Mice were injected with lipopolysaccharide (LPS,1 mg/kg) 2 h after the pretreatment of EMD638683 (10 mg/kg) or phosphate buffered saline (PBS) as control.At the time points of 3 and 24 h,pro-inflammatory cytokines (interleukin [IL]-6,IL-12 and tumor necrosis factor [TNF]-α) in serum were measured using enzymelinked immunosorbent assay (ELISA).Livers and lung were harvested at 6 h and 24 h after the injection of LPS,embedded by optimum cutting temperature (OCT) and then stained with hematoxylin and eosin (HE).Peripheral blood mononuelear cell (PBMC) were isolated and stimulated by LPS with or without the pretreatment of EMD or LY294002.Cytokines (IL-6,IL-12 and TNF-α) were measured using ELISA.IKKα/β,IKBα and nuclear factor (NF)-κB p65 were detected by Western bolt.Data were analyzed by one way analysis of variance.Results In the model of LPS-induced endotoxin sepsis,inhibition of SGK1 induced secretion of pro-inflammatory cytokine (IL-6 [t=3.007,P＜0.05],IL-12[t=4.413,P＜0.05] and TNF-α[t=5.403,P＜0.05]),increased inflammatory cells infikration into the liver and lung within 6 h,and induced serious multiple organ damage with collapse of alveoli and fatty degeneration of liver.After 24 h,pharmacological inhibition of SGK1 with EMD638683 increased proinflammatory cytokine (IL-6 [t=18.540,P＜0.01],IL-12[t=16.520,P＜0.01] and TNF-α[t=34.880,P＜0.01]) production in human PBMC upon LPS stimulation and inhibited the phosphorylation of IKKα/ β/IKBα and nuclear factor (NF)-κB p65.Conclusions SGK1 suppresses the toll like receptor 4 mediated inflammatory responses via NF-κB.

P53 gene (exon7～8) mutatins and p53 proteins and HPV 6，11，16，18-DNA were examined in 49 cervical carcinoma by immunohistochemistry, polymerase chain reaction (PCR) and single strand conformation polymorphism (SSCP) in order to investigate their role and mutual relation and clinical significance in the onco genesis of cervical carcinoma. The results showed that first, p53 proteins posit ive rate was 48.98％, and not outstandingly related to the differentiation and the invasive degree of cervical carcinoma(P＞0.05); the defects of P53 gene (exon7～8) were not found but P53 (exon7～8) mutations were detected in 7 of 49(14.29％) cervical carcinoma; then, HPV16-DNA positive rate was much higher than HPV6,11,18-DNA positive rate respectively(P＜0.001),and the different HPV-DNA was simultaneously tested in one cerv ical carcinoma; last, not all cases of P53 mutations had p53 proteins posi tive, but the cases of P53 mutations and p53 proteins negative certainly had HPV infections, and HPV positive cases were much more than its negative one in the cases of p53 proteins positive(P＜0.001). These results proved that the oncogenesis of cervical carcinoma is mainly associated with HPV16 infections, and second related to P53 (exon7～8) mutations. p53 proteins positive results from P53 mutations or/and HPV infections in cervical carc inoma.

Colonic Diseases ; Hepatitis C, Chronic ; Humans ; Interferons ; Ischemia

Objective To obtain the disulfide stabilized Fv fragment (dsFv) against N-terminal fragment of human lip polysaccharide binding protein (NH-LBP) and to identify its biological vitality. Methods The disulfide stabilized Fv fragment antibody (dsFv) was obtained after the inclusion bodies of dsFvVH and dsFvVL had been refolded and purified. Then the characteristics of dsFv were determined in vitro by ELISA and by detecting the secretion of tumor necrosis factor alpha (TNF-?) in rats. Results There was 2.1 mg protein of dsFv obtained. dsFv had good combination with NH-LBP and could restrain inflammatory reaction caused by lip polysaccharide (LPS) in vivo. Conclusion It is feasible to get dsFv against NH-LBP by respective expression of VL and VH. The partial inhibition of the biological function of LBP by dsFv is a new way to restrain the over-inflammatory reaction in vivo.

Objective To reconstruct the proteinic sequence of human cathelicidin LL-37 to increase the bactericidal activity of LL-37 and to express the reconstructed LL-37 (rLL-37) in bacterium. Methods The two dimensional structure, three dimensional structure and chemical characteristic of LL-37 were analyzed by Soft Ware Anthepro 5.0 and SWISS-MODEL. Without the three dimensional changes of LL-37, some negative amino acids of human cathelicidin LL-37 were replaced by positive amino acids and the positive charge of LL-37 was increased. According to the proteinic sequence changes of rLL-37, the DNA sequence of rLL-37 was reconstructed by Touch-Down PCR and recombined with vector pET-28a (+), thus rLL-37 was expressed in E.coli. BL21 (DE3) by the induction of IPTG and was purified by chromatography. Results Glu~ 16 , Asp~ 26 , Glu~ 36 of LL-37 were replaced by Gln~ 16 , Asn~ 26 , Gln~ 36 and the static charge of LL-37 was increased from +5.8 to +9.0 at pH 7.4. The DNA sequence of rLL-37 was reconstructed and inserted into vector pET-28a (+), the rLL-37 was expressed in E.coli. BL21 (DE3) and purified by strong cation exchange supports Macro-Prep High S successfully. The rLL-37 was proved by the means of inhibitory zone to be able to kill Gram-negative bacteria and Gram-positive bacteria. Conclusion It is feasible to reconstruct human cathelicidin LL-37 and express the protein in bacteria by fusion, which make it possible to produce more rLL-37 and study its biological function deeply.