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

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 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.

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.

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

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

This study was aimed to observe the analgesia of curing-injury Cataplasma and discuss the Nav1 . 7 expression in dorsal root ganglion ( DRG ) in model rats with formaldehyde-induced inflammatory pain . A total of 36 Sprague-Dawley rats were divided into three groups, which were the blank control group (n = 12), model group ( n = 12 ) , and treatment group ( n = 12 ) . The blank control group was without any treatment . The model group was injected with 0 . 1 mL 5% saline formalin on the left rear foot . The treatment group was applied with curing-injury Cataplasma on the left rear foot 24 h before the injection of 0 . 1 mL 5% saline formalin in the establishment of animal model . The behavior reactions to pain of model rats were observed . Dubuisson score was recorded and compared . Meanwhile , L3-6 DRG was collected from rats in each group . The expres-sion of Nav1 . 7 was detected by real-time quantitative PCR and western blot . The results showed that the pain reaction integral in the treatment group was lower than the model group ( P < 0 . 05 ) . Results from the real-time quantitative PCR showed that the relative expression of Nav1 . 7 mRNA in the model group was more than the treatment group . And the relative expression of Nav1 . 7 mRNA in the treatment group was more than the blank control group . There was significant difference among three groups ( P < 0 . 05 ) . There was no statistical difference at the three time points within three groups. Results from the western blot showed that the relative expression of Nav1 . 7 in the model group was more than the treatment group . And the expression of Nav1 . 7 in the treatment group was more than the blank control group . There was significant difference among three groups (P < 0.05). There was no statistical difference at the three time points within three groups. It was concluded that the curing-injury Cataplasma can alleviate inflammatory pain response in rats, and have certain analgesia effect . Meanwhile , it can influence Nav1 . 7 expression in DRG in model rats with formaldehyde-induced inflam-matory pain .

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.

Objective To explore the CT findings of benign and malignant pancreatic neuroendocrine tumors and improve its diagnostic accuracy.Methods The clinical information and enhanced CT findings of 96 cases with pathologically-proved pancreatic neuroendocrine tumors were retrospectively reviewed.The CT findings were evaluated by several factors,which included tumor size,morphology,location,internal composition,calcification,separation,bile duct and pancreatic duct dilation and CT value.Results All cases were divided into benign or malignant according to pathological grades,and benign group involved 40 cases with 41 lesions,while malignant group involved 56 cases with 59 lesions.The size of malignant lesions was significantly larger than that of benign lesions (median size 6.0 cm vs 2.2 cm),the shape of the lesions was irregular,and was mainly cystic solid,and mottling,curve shape,clumps calcification was present,then the bile duct and pancreatic duct was mild to moderately dilated,and the difference between the two groups was statistically significant (P ＜0.05).But the difference of tumor location,separation was not significant.45.76％ (27/59) of the malignant lesions reached the peak value in arterial phase,and 44.07％ (26/59) reached the peak value in venous phase;while 68.29％ (28/41) of the benign lesions reached the peak value in arterial phase,and 31.71％ (13/41) reached the peak value in venous phase.The CT values of malignant lesions in plain CT scanning,arterial phase,venous phase,balance phase were (39.02 ±7.53),(121.20 ± 54.73),(125.25 ± 40.77),(101.41 ± 28.68) Hu,while they were (41.49 ± 8.59),(144.73 ± 53.95),(157.05 ±44.72),(121.02 ±29.80) Hu in benign group.In plain CT scanning,the difference of CT value between malignant and benign lesions was not significant;but in the enhanced phase,the CT value of malignant lesions was significantly lower than that of benign lesions,and the difference was statistically significant (P ＜ 0.05).Conclusions The lesion with its size ≥ 3.0 cm,irregnlar morphology,cystic necrosis,calcification,pancreatic and bile duct dilatation is suggestive of malignancy tumor.The average CT values of malignant group are lower than those of the benign group in arterial,venous and balance phases.

Objective:To investigate the effect and mechanism of exosomes derived from human-induced pluripotent mesenchymal stem cells (iMSC-Exos) on alveolar macrophages (AM) pyroptosis.Methods:The exosomes in the culture supernatant of human-induced pluripotent mesenchymal stem cells (iMSC) were extracted by rotating ultrafiltration, and the extracted exosomes were identified by transmission electron microscopy, Western blotting and high-resolution adjustable resistance pulse. The rat alveolar macrophage cells (NR8383 cells) were cultured in vitro and the logarithmic growth phase cells were divided into three groups: the control group was added with an equal volume of phosphate buffered saline (PBS) in the AM supernatant; in LPS/ATP group AM cells were stimulated with 500 μg/L LPS for 23 hours and then 5 mmol/L ATP was added for 1 hour to induce pyrolysis; iMSC-Exos group was incubated with AM and 100 mg/L iMSC-Exos for 3 hours before giving LPS and ATP. The cytotoxic activity was detected by cell counting kit-8 (CCK-8) and lactate dehydrogenase (LDH) analysis, the apoptosis and the expression of caspase-1 were observed by immunofluorescence, the levels of inflammatory factors interleukins (IL-1β and IL-18) released by AM were detected by enzyme linked immunosorbent assay (ELISA), the NOD-like receptor protein 3 (NLRP3) inflammasome pathway and the expression level of pyroptosis related protein gasdermin D (GSDMD) were detected by Western blotting. Results:The extracted exosomes were observed by transmission electron microscopy as round vesicles, expressing exosomal markers CD63 and CD9 showed by Western blotting, high-resolution adjustable resistance pulse showed the average diameter of the particles was 130 nm, and could be uptaken by AM. Compared with the control group, the cell activity decreased [(0.56±0.05)% vs. (1.06±0.07)%, P < 0.01], the release of necrotic substance LDH increased (U/L: 1 218.86±22.73 vs. 188.30±1.61, P < 0.01), the expression levels of inflammatory factors increased [IL-1β (ng/L): 958.91±32.78 vs. 194.63±5.14, IL-18 (ng/L): 870.89±21.86 vs. 288.85±24.48, both P < 0.01], and the apoptosis rate [(55.35±6.19)% vs. (12.01±1.32)%, P < 0.01] and caspase-1 expression (fluorescence intensity: 41.06±3.65 vs. 2.80±0.54, P < 0.01) elevated in the AM after LPS/ATP stimulation, suggesting that LPS combined with ATP successfully induced alveolar pyroptosis. Compared with the LPS/ATP group, AM pretreated with iMSC-Exos showed increased cell viability [(0.81±0.05)% vs. (0.56±0.05)%, P < 0.01], decreased LDH secretion (U/L: 535.05±42.55 vs. 1 218.86±22.73, P < 0.01), decreased expression of inflammatory factors [IL-1β (ng/L): 381.82±19.50 vs. 958.91±32.78, IL-18 (ng/L): 533.77±31.54 vs. 870.89±21.86, both P < 0.01], and decreased apoptosis rate [(19.74±2.96)% vs. (55.35±6.19)%, P < 0.01] and caspase-1 expression (fluorescence intensity: 12.16±1.31 vs. 41.06±3.65, P < 0.01). At the same time, the expression of NLRP3 inflammasome pathway [NLRP3 protein (NLRP3/β-actin): 0.62±0.06 vs. 1.89±0.11; cleaved caspase-1 protein (cleaved caspase-1/β-actin): 0.42±0.07 vs. 1.22±0.17, both P < 0.01] and pyrolysis-related protein was significantly inhibited [GSDMD protein (GSDMD/β-actin): 0.57±0.05 vs. 1.22±0.05, P < 0.01]. Conclusion:iMSC-Exos successfully reversed the AM pyroptosis and inflammatory factor expression induced by LPS/ATP, which may be due to the targeted inhibition of NLRP3 inflammasome pathway, suggesting that iMSC-Exos can exert anti-inflammatory effects by inhibiting the pyrolysis of AM.