Objective To investigate the effects of L-thyroxine(LT4) on serum lipid and bone metabolism in aged patients with subclinical hypothyroidism (SH). Methods Seventy-eight patients with diagnosed SH were randomly assigned to receive LT4 (therapy group) or placebo(control group) for one year. Mean dose for LT4 replacement was (80.1?37.2) ?g/d. The changes of clinical symptoms, body mass index (BMI) and serum levels of total cholesterol(TC), triglycerides(TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol(LDL-C), apolipoprotein A 1(ApoA 1), apolipoprotein B 100 (ApoB 100 ), calcium ion(Ca 2+ ), phosphorus ion (P 3+ ) and alkaline phosphatase (AKP) and bone mineral density (BMD) were measured before and after therapy. Results After LT4 therapy, clinical symptoms were ameliorated. BMI and serum TC, TG, LDL-C and ApoB 100 levels were significantly reduced 〔(24.6?3.1 vs 23.3?2.2),(5.65?1.42 vs 4.43?0.81),(1.81?0.66 vs 1.43?0.40),(3.64?0.84 vs 2.68?0.63)mmol/L,(0.98?0.22 vs 0.83?0.21)g/L,P

Objective; To express and purify HCA518 protein and prepare its monoclonal antibody ( McAb). Methods: The HCA518 protein was expressed with gene recombinant technique in prokaryotic system and purified with nickel chelate nitrilotriacetic acid(Ni-NTA) affinity chromatography column. Hybridoma cell lines that secreted anti-HCA518 McAb were established by cells fusion and screened by enzyme linked immunosorbent assay( ELISA). The specificity of anti-HCA518 McAb wa3 identified by Western blot assay. The HCA518 protein in tumor cells was stained by immunoflourescence assay. Results: Rcombinant HCA518 protein was expressed with a purity of 98%. Two hybridoma cell lines was selected and anti-HCA518 McAb was purified from mice ascites. The titers of anti HCA518 McAb in ascites were 1?10-4 and 5?10-4 respectively. The antibody belonged to IgG2b subtype and IgM. Anti-HCA518 McAb specifically reacted with recombinant HCA518 protein and tumor cells'nuclear protein (P100). The HCA518 protein was mainly located in cell nucleus. Conclusion: Stable hybridoma cell lines that secreted anti-HCA518 McAb have been established and anti HCA518 McAb was prepared with high specificity. It has important significance for detecting HCA518 protein in tumor tissues and determining malignant proliferation status of tumor cells and predicting its prognosis.

The gene encoding thermostable lactate dehydrogenase (Tm-LDH) was cloned into the plasmid pHsh from Thermotoga maritima, and expressed in Escherichia coli JM 109. The recombinant protein was purified to homogeneity by a simple step, heat treatment. The recombinant enzyme had a molecular mass of 33 kDa. The optimal temperature and pH of Tm-LDH were observed 95 degrees C and 7.0. The purified enzyme had a half-life of 2 h at 90 degrees C, and exhibited better stability over a pH range from 5.5 to 8.0. The K(m) and V(max) values were 1.7 mmol/L, 3.8 x 10(4) U/mg of protein for pyruvate, and 7.2 mmol/L and 1.1 x 10(5) U/mg for NADH, respectively. The expression of Tm-LDH in T7 system could not obtain high efficiency, but it has been soluble over-expression in pHsh system and reached 340 mg/L. The superior stability and productivity of Tm-LDH will lay the foundation of its industrial-scale fermentation and application in the NAD regeneration.
Cloning, Molecular ; Enzyme Stability ; Escherichia coli ; metabolism ; L-Lactate Dehydrogenase ; biosynthesis ; Molecular Weight ; Recombinant Proteins ; biosynthesis ; Temperature ; Thermotoga maritima ; enzymology

Objective To find an effective CT-MRI image fusion protocol in brain tumor by analyzing the registration accuracy of different methods. Methods The simulation CT scan and MRI T1 WI imaging of 10 brain tumor patients obtained with same position were registered by Tris-Axes landmark 、Tris-Axes landmark + manual adjustment、 mutual information and mutual information + manual adjustment method. The clinical tumor volume (CTV) were contoured on both CT and MRI images respectively. The accuracy of image fusion was assessed by the mean distance of five bone markers ( d1-5 ), central position of CTV ( dCTV ) the percentage of CTV overlap ( PCT-MRI ) between CT and MRI images. The difference between different methods was analyzed by Freidman M non-parameter test. Results The difference of the means d1-5 between the Tris-Axes landmark、Tris-Axes landmark plus manual adjustment、mutual information and mutual information plus manual adjustment methods were 0. 28 cm ±0. 12 cm, 0. 15 cm ±0.02 cm, 0. 25 cm± 0. 19 cm, 0. 10 cm ± 0. 06 cm, ( M = 14. 41, P = 0. 002 ). the means dCTV were 0. 59 cm ± 0. 28 cm,0. 60 cm± 0. 32 cm, 0. 58 cm ± 0. 39 cm, 0. 42 cm± 0. 30 cm( M = 9. 72, P = 0. 021 ), the means PCT-MRI were 0.69% ±0. 18%, 0.68% ±0. 16%, 0.66% ±0. 17%, 0.74% ±0. 14% (M = 14.82,P=0.002),respectively. Conclusions Mutual information plus manual adjustment registration method was the preferable fusion method for brain tumor patients.

Objective To evaluate the efficacy of port catheter system(PCS)placement in pulmonary artery via percutaneous subclavicle vein treatment for multiple metastatic tumor in the two lungs and discuss the PCS technique.Methods Fifteen multiple metastatic tumor patients(13 hepatocellular carcinomas,one mandible grand adenocarcinoma,one oral bottom squamous carcinoma)were carried out with pulmonary artery PCS placement by way of percutaneous subclavicle vein.FPA/FPM/GP chemotherapy scheme were introduced every 4～6 weeks.Results The success rate of PCS placement technique was 93.3%(14/15).One case failed.Percutaneous subclavicle veins were performed 14 cases in right side and 1 in left one.Following up 2～43 months,2～7 chemotherapy cycles(mean 5 cycles)were accomplished,and the clinical CR and PR were achieved in 1 and 3 cases respectively with clinical efficacy rate 28.6%(4/14).Major side reaction was late wound healing in 1 case.Conclusion PCS placement in pulmonary artery treatment for multiple metastatic tumor in the two lungs is effective,and mastering operation technique is the key for increasing operation suc- cess rate.

Background Studies showed that activation of microglia-derived nicotinamide adenine dinucleotide phosphate (NADPH) oxidase plays a key role in the neurodegenerative diseases and neural cell death in central nervous system.The effect of NADPH on cone degeneration have been determined in rd rats,its role in rod degeneration is relatively less studied.Objective This study was to study the expression of NADPH oxidase in the retinal degenerative process in rd mice and further explore its role in the photoreceptor degeneration.Methods rd Mice at postnatal day 8 (P8),P10,P12,P14,P16 and P18 were collected.The mice were sacrificed,and retinal sections,RNAs and proteins were prepared in above-mentioned time points.The expressions of the gp91 phox,a major subunit of NADPH oxidase,in transcript level and protein level in the retinas were semi-quantitatively detected by real-time PCR and Western blot respectively.Expression of gp91phox was localized in the rd retinas as ageing by immunohistochemstry,and the co-expression of gp91phox with CD11b,a specific marker of microglial cells,was assayed by immunofluorescent double labeling.The C57BL/6N mice were served as controls.The use and care of the animals complied with the Guideline of ARVO.Results Real-time PCR showed that gp91phox mRNA was not expressed in the retinas of C57BL/6N mice.Gp91phox mRNA was found to have less expressed in retinas of P8 rd mice.With aging,the expression level of gp91phox mRNA (gp91phox mRNA/β-actin) in rd mouse retinas was gradually increased with the highest level in P14 mice(1.136±0.370).A significant difference was seen in the gp91 phox mRNA expression among various groups of mice (F=17.81,P =0.00),and gp91phox mRNA expression was significantly elevated in P10,P12,P14,P16 and P18 rd mice compared with P8 rd mice(all at P＜0.05).The expression level of gp91phox protein (A value) in the retinas presented with a similar trend in the rd mice,with a significant difference among the various ages of rd mice and C57BL/6N mice (F =354.00,P＜0.01).The expression level of gp91 phox protein was increased in the rd mice in comparison with the C57BL/6N mice (all at P＜0.05).Immunochemistry revealed that the positive response cells for gp91phox increased in the inner layers of retinas in P10 rd mice and peaked in P14 mice.Immunofluorescent double labeling exhibited that gp91phox were seen to present a co-expression with CD11b,showing an orange fluorescence.Conclusions Expression of NADPH oxidase in the rctinas in the rd mice up-regulates and is parallels to the microglial activation and photoreceptor degeneration,suggesting that NADPH oxidase plays a role in the retinal dystrophy associated with microglial activation.

Background and purpose: Arginase-1 (Arg-1) is an enzyme involved in the urea cycle. Research has shown that changed expression of Arg-1 plays an important role in the cellular metabolism and growth. The purpose of this research was to investigate the expression of Arg-1 in hepatocellular carcinoma (HCC) and to analyze its correlation with clinicopathological features. Methods: The expression of Arg-1 protein and mRNA in 31 samples of HCC, paracancerous liver tissues and 12 samples of normal liver was detected by Western blot and reverse transcription-polymerase chain reaction (RT-PCR). The expression profiles of Arg-1 at protein level in 158 samples of HCC and paracancerous liver tissues were detected with high-throughput tissue microarray technique and immunohistochemistry. The relationships between Arg-1 expression and clinicopathological features were also analyzed. Results:The expression of Arg-1 mRNA and protein was signiifcantly decreased in HCC compared with the paracancerous liver tissues and normal liver tissues (F=57.83, 160.89; all P<0.01). The expression of Arg-1 in HCC was related to differentiation degree (F=10.41, 30.03; all P<0.01). And with tumor differentiation decreased, the expression level down-regulated. Immunohistochemistry revealed that Arg-1 protein was mainly located in the cytoplasm and nuclear. The expression rates of Arg-1 were 88%, 98.7%and 100%in HCC, paracancerous tissues and normal liver tissues, respectively. Statistical analysis revealed that Arg-1 protein staining rate was signiifcantly lower in tumor tissues than that in paracancerous tissues (χ2=14.7416, P<0.01) and normal liver tissues (χ2=4.1415, P<0.05). The Arg-1 expression was correlated with differentiation degree of HCC, vascular invasion and recurrence after operation (χ2=22.8459, 10.2639, 10.6368 respectively;all P<0.05), but not correlated with age, gender, the hepatitis B virus, the level of serum AFP, liver cirrhosis, diameter of tumor and tumor number. Conclusion:The expression level of Arg-1 is much lower in HCC than that in the paracancerous liver and normal liver. Moreover, Arg-1 expression level is closely related to tumor differentiation degree, metastasis and relapse. These data demonstrate that Arg-1 may play a negative role in the development of HCC.

Animals ; Chickens ; physiology ; Eating ; drug effects ; Injections, Intraventricular ; methods ; Neuropeptide Y ; administration & dosage ; pharmacology

Aged ; Humans ; Kidney Neoplasms ; pathology ; Male ; Neoplasm Metastasis ; Paranasal Sinus Neoplasms ; secondary

Objective T o explore the application value of postm ortem com puted tom ography (C T ) an-giography on diagnosis of coronary atherosclerotic stenosis degree. Methods B ased on the previous ex-perim ental results, the postm ortem C T angiography device of hum an isolated heart w as im proved. D iffer-ent coronary atherosclerotic stenosis degree of sudden death cases w as selected. B efore the cardiac anatom y, hearts w ere rem oved out com pletely and C T angiography w as perform ed im m ediately. T he C T angiography results w ere com pared w ith histopathological findings. M eanw hile, the advantages and disad-vantages of the angiography device before and after im provem ent w ere com pared. Results T he im proved angiography device of isolated heart could get better im aging results. T he postm ortem C T angiography results had high consistency w ith the histopathological findings on diagnosis of coronary atherosclerotic stenosis degree. A nd the coronary artery lesions could be revealed m ore objectively and vividly by 3D reconstruction technology. H ow ever, C T angiography could only be used to exam ine the pathological changes of blood vessels, w hich m ight have som e lim itations on the diagnosis of cause of death. Con-clusion Postm ortem C T angiography can be used as an additional m ethod for the conventional autopsy in the cases of coronary atherosclerosis.