Biological Properties of 3D-Printed Zirconia Implants with p-Cell Structures.

Research on 3-dimensional (3D) printed porous zirconia-based dental implants is still in its infancy. This study aimed to evaluate the biological responses of novel zirconia implants with p-cell structures fabricated by 3D printing. The solid zirconia samples exhibited comparable density, 3-point flexural strength, and accelerated aging properties compared to specimens prepared previously by conventional methods. Cell-based experiments showed that the p-cell structure promoted cell proliferation, adhesion, and osteogenesis-related protein expression. Mechanical tests showed that both p-cell and control implants could withstand a torque of 35 Ncm without breaking. The mean maximum breaking loads of p-cell and control implants were 1,222.429 ± 115.591 N and 1,903.857 ± 250.673 N, respectively, which were much higher than the human physiological chewing force and human mean maximum occlusal force. An animal experiment showed that the bone trabeculae around the implants were significantly thicker, more numerous, and denser in the p-cell group than in the control group. This work could provide promising guidance for further exploring 3D printing techniques for porous zirconia bionic implants in dentistry.

Immunohistochemical investigation of the correlation between LIM kinase 1 expression and development and progression of human ovarian carcinoma.

OBJECTIVES: LIM kinase 1 (LIMK1) is implicated in cellular mechanisms regulating tumour cell invasion and may be involved in ovarian carcinoma progression. This retrospective study, therefore, investigated expression of the LIMK1 gene in primary ovarian tumour tissue samples and evaluated the correlation between LIMK1 gene expression and progression of ovarian carcinoma. METHODS: LIMK1 protein levels were detected by immunohistochemistry in ovarian tissue samples from 57 patients with primary ovarian epithelial tumours (benign, n = 13; borderline, n = 14; carcinoma, n = 30) and 10 patients with normal ovaries. LIMK1 protein levels were evaluated by calculating the product of the scores for stain intensity and percentage of cells stained. RESULTS: There was a significant correlation between increasing LIMK1 protein levels and increasing disease severity, from normal ovarian tissues through benign and borderline tumours to ovarian carcinoma. There was also a significant correlation between increasing LIMK1 protein levels and increasingly poor levels of differentiation of ovarian carcinoma. CONCLUSIONS: LIMK1 is associated with the development of ovarian cancer and with the level of tumour differentiation in patients with ovarian carcinoma.

Increase of Mcm3 and Mcm4 expression in cervical squamous cell carcinomas.

PURPOSE: To examine the relevance of expression of two proteins essential for DNA replication initiation, Mcm3 and Mcm4, in cervical squamous cell carcinoma (CSCC). METHODS: 53 cases of cervical squamous cell carcinoma, 35 cervical intraepithelial neoplasias (CIN) grade 2-3, 11 CIN I and 26 normal controls were studied. Immunohistochemistry was performed with anti-Mcm3 and anti-Mcm4 antibodies. RESULTS: We found Mcm3 and Mcm4 protein expression had a tendency to be stronger from control to CSCC. Both Mcm3 and Mcm4 were significantly upregulated in squamous cervical carcinoma compared to the control, CIN grade 1 and grade 2-3 (p < 0.001), and Mcm3 expression was correlated with CSCC cell differentiation. However there were no independent prognosis correlations between Mcm3 and Mcm4 and clinicopathological parameters including age, stage, tumor size, invasive depth and lymph node metastasis. CONCLUSIONS: Mcm3 and Mcm4 were highly expressed in CSCC, and these two proteins might be useful as biomolecular markers in clinical diagnosis.

The expression of interleukin-10 in patients with primary ovarian epithelial carcinoma and in ovarian carcinoma cell lines.

This study determined interleukin-10 (IL-10) expression in patients with ovarian carcinoma and in ovarian carcinoma cell lines, and investigated its clinical significance in the development and progression of ovarian carcinoma. Expression of IL-10 in ovarian carcinoma, benign ovarian tumour, normal control tissues and ovarian carcinoma cell lines was detected by immunohistochemistry and Western blot analysis. IL-10 concentrations in sera and ascites from patients with ovarian carcinoma, in sera from patients with benign ovarian tumour and normal controls, and in supernatants of ovarian carcinoma cell line cultures were measured by enzyme-linked immunosorbent assay. The tissue level of IL-10 in ovarian carcinoma was significantly higher than in benign ovarian tumour and normal controls. IL-10 expression was detectable in cell lysate and supernatant from ovarian carcinoma cell lines. In patients with ovarian carcinoma the IL-10 level in ascitic fluid was significantly higher than in sera, and the serum IL-10 level in ovarian carcinoma was significantly higher than in benign ovarian tumour and normal controls. Ascitic IL-10 levels in ovarian carcinoma were significantly correlated with disease stage but not cytological grade. These results suggest that ovarian carcinoma cells are able to synthesize and secrete IL-10, which probably assists in promoting the development and progression of ovarian carcinoma.

Annual dynamics and origins of the odorous compounds in the pilot experimental area of Lake Dianchi, China.

Lake Dianchi is the sixth largest freshwater lake in China. The pilot experimental area (6 km2) in Lake Dianchi is one of the most severely polluted areas in the lake with heavy cyanobacteria blooms. During June 2002 to May 2003, the algal composition and number, and odorous compounds were identified monthly and monitored in the area. Meanwhile, physicochemical parameters such as total phosphorus (TP), total nitrogen (TN), 5-day biochemical oxygen demand (BOD5), chemical oxygen demand (COD(Mn)), dissolved oxygen (DO), pH, transparency, temperature and chlorophyll a (chla) were determined. Four odorous compounds: 2-methylisoborneol (MIB), geosmin, beta-cyclocitral and beta-ionone were found in the lake water. Both the concentration of particulate beta-cyclocitral and that of beta-ionone correlated significantly with the annual variation of Chla, biomass of total phytoplankton, cyanobacteria and Microcystis. The concentration of particulate MIB correlated significantly with the biomass of Oscillatoria, while the concentration of particulate geosmin correlated significantly with the biomass of Anabaena. Off-flavour in the pilot area was found to be caused by a combination of beta-cyclocitral- and beta-ionone-producing Microcystis, MIB-producing Oscillatoria and geosmin-producing Anabaena. beta-ionone, MIB and geosmin contributed collectively to the odour intensity in the lake water.

Neurotransmitter dopamine applied in electrochemical determination of aluminum in drinking waters and biological samples.

It was demonstrated that the decrease of the differential pulse voltammetric (DPV) anodic peak current of dopamine (3,4-dihydroxyphenylethylamine, DA) was linear with the increase of aluminum (Al) concentration. Under optimum experimental conditions (pH 4.6, 1.2 x 10(-3) M DA, and 0.04 M NaAc-HAc buffer solution), the linear range is 4.0 x 10(-7)-8.0 x 10(-5) M, the detection limit is 1.4 x 10(-7) M, and the relative standard deviation for 4 x 10(-5) M Al(III) is 3.5% (n=8). Many foreign species, especially some low-molecule-weight biological molecules, were chosen for interference testing. The proposed method was applied to the determination of Al in biological samples such as synthetic renal dialysate, Ringer's solution, human blood, cerebrospinal fluid of a patient, and urine of a diabetic patient. The corresponding recoveries were generally between 95 and 105%. The basic principle of the method was determined by examining Al complexed with DA. This results in the blockage of the electroactive sites on DA, followed eventually by the reduction of the electrochemical response of DA. This result was verified by examining the behavior of DA, both in the presence and absence of Al, using electrochemical, UV-Vis, Raman, and (13)C NMR spectroscopic methods.

Speciation of aluminium(III) in natural waters using differential pulse voltammetry with a pyrocatechol violet-modified electrode.

A differential pulse voltammetric (DPV) procedure is proposed for the speciation of aluminium in natural waters using Pyrocatechol Violet chemically modified electrodes (PCV-CMEs). This novel speciation idea is based on the selective determination of different AlIII forms under two pH conditions. The labile monomeric Al fraction (mainly inorganic Al) is analysed at pH 4.8 (0.20 mol dm(-3) NaOAc-HOAc) and the total monomeric Al fraction is analysed at pH 8.5 (0.20 mol dm(-3) NH3.H2O-NH4Cl). The difference is thought to be caused by the weak competition ability of PCV to sequester AlIII from AlIII-natural organic matter complexes. This sensitive and simple speciation method has been applied successfully to aluminium speciation in natural waters sampled from different regions of China. Five fractions are measured directly or indirectly: (i) labile monomeric Al; (ii) total monomeric Al; (iii) acid reactive Al; (iv) non-labile monomeric Al; and (v) acid soluble Al. The results are in satisfactory agreement with those obtained by Driscoll's 8-hydroxyquinoline extraction-ion exchange method.

Trace analysis of aluminum in natural waters with rubeanic acid by adsorption chronopotentiometry.

The determination of trace levels of aluminum in natural waters with rubeanic acid (RA) by adsorption chronopotentiometry is developed in this paper. Optimum experimental conditions include an accumulation potential of -0.40 V, accumulation time of 60 s, and a RA concentration of 6x10(-6) M in 0.2 M NaAc-HAc buffer solution (pH 4.6). The response is linear over the 1x10(-8) approximately 4x10(-7) M concentration range. The detection limit is 5.6x10(-9) M and the relative S.D. (at the 3x10(-7) M level) is 2.6%. Possible interferences are evaluated. The method has been applied to the determination of trace levels of Al in various real samples. Direct determination of toxic forms of Al in surface waters by this technique is also explored.

Effects of a single drip infusion of lipo-prostaglandin E1 on vibratory threshold in patients with diabetic neuropathy.

We evaluated acute effects of prostaglandin E1 encapsulated in lipid microspheres (lipo-PGE1), in 14 type 2 diabetic patients with neuropathy. Nerve conduction velocity (NCV), vibratory threshold (VT), skin temperature and subjective symptoms were evaluated for 24 hours after a single drip infusion of lipo-PGE1. In 6 of the 14 patients, the decrease in VT (P < 0.05) more than 50% measured by an SMV-5 vibrometer was observed at 6 hours after the infusion (responders). Subjective symptoms were improved (P < 0.05) in 5 of the 6 responders, whereas it improved in only 1 of the 8 patients who had no change in VT (nonresponders). NCV did not change (P > 0.05) either in the responders or in the nonresponders by the infusion. The responders had less impairment in VT and milder retinopathy and nephropathy than the nonresponders (P < 0.05). Our results demonstrate that vibratory sensation and subjective symptoms can be improved by a single infusion of lipo-PGE1 in type 2 diabetic patients with mild neuropathy.

The effects of goshajinkigan, a herbal medicine, on subjective symptoms and vibratory threshold in patients with diabetic neuropathy.

Goshajinkigan, a herbal medicine, has long been used in Japan to alleviate the subjective symptoms of diabetic neuropathy; however, its effects have not been confirmed objectively. We evaluated its effects on subjective symptoms and on vibration sensation in patients with diabetic neuropathy. The oral administration of 7.5 g/day of Goshajinkigan for 3 months (treatment period) relieved subjective symptoms of numbness in 9 of 13 patients. When the drug was discontinued for 2 months as a washout period, the subjective symptom worsened in 7 of 13 patients. Chi-square analysis revealed significant effects of Goshajiniagan on subjective symptoms (P < 0.001 for numbness and P < 0.05 for cold sensation). Vibration sensation was evaluated by measuring vibratory threshold using an SMV-5 vibrometer. There were significant changes in vibratory thresholds by paired t-test (P < 0.05) both in the upper and the lower extremities during the treatment and washout periods. Chi-square analysis also revealed a significant effect of Goshajinkigan on vibratory threshold (P < 0.01). There was no significant change in glycosylated hemoglobin as a whole during the study. These observations confirm that Goshajinkigan relieves subjective symptoms and demonstrate that it improves vibration sensation in patients with diabetic neuropathy.

Variant forms of glucokinase gene in Japanese patients with late-onset type 2 diabetes.

We have applied the technique of single-strand conformation polymorphism analysis to detect mutations of the glucokinase gene in 50 Japanese patients with late-onset type 2 diabetes and in 50 normal Japanese subjects. Out of the 50 patients with late-onset type 2 diabetes, we observed three kinds of variant patterns: one in exon 1b, one in exon 4, and one in exon 5. The incidence of these patterns was one in exon 1b, two in exon 4 and one in exon 5. Direct sequencing of exon 1b and exon 5 revealed mutations in intron areas at the 12th nucleotide downstream from the 5' splice points in two cases. Direct sequencing of exon 4 revealed a heterozygous silent mutation, CCC[Pro]-->CCG[Pro] at codon 145. In contrast, 50 normal Japanese subjects showed no variant patterns in any exons. Our results showed that although 8% (4 out of 50) of Japanese patients with late-onset type 2 diabetes have variant forms of the glucokinase gene, none is expected to cause apparent qualitative changes in glucokinase. We think that the frequency of mutations of the glucokinase gene which could cause qualitative change is very low in Japanese patients with late-onset type 2 diabetes.

Change in bone mineral density in patients with hyperthyroidism after attainment of euthyroidism by dual energy X-ray absorptiometry.

Because of the previous controversial findings in studies of bone mineral density in patients with hyperthyroidism with older methodologies, we assessed bone mineral density in 15 Japanese patients with Graves' disease (8 males and 7 females) before and after treatment using dual energy X-ray absorptiometry (DEXA). Bone mineral density of the lumbar vertebrae and the femur, and thyroid function, and several metabolic parameters were measured before treatment and again after patients achieved a euthyroid state following treatment with methimazole for 4 to 20 months (mean 10.6 months). The bone mineral density of patients was calculated as the percentage of the mean value (%BMD) in an age- and sex-matched control group, and correlations between the changes in bone mineral density and metabolic parameters before and after treatment were investigated. The %BMD of vertebrae in patients with Graves' disease before treatment was 89.7% of that found in the normal population. When patients became euthyroid after treatment, %BMD increased significantly to 94.9%, although it still remained below the control level. TSH receptor antibody, osteocalcin, and alkaline phosphatase were elevated before treatment, but decreased significantly after treatment. The change between pre- and posttreatment TSH receptor antibody was negatively correlated with the change in bone mineral density. In conclusion, these findings suggest that bone mineral density is decreased in patients with Graves' disease and that successful treatment of hyperthyroidism results in a significant increase in bone mineral density within a short period of time. Furthermore, TSH receptor antibody is a useful marker of changes in bone metabolism in this group of patients.

Bone mineral density in patients with hyperthyroidism measured by dual energy X-ray absorptiometry.

OBJECTIVE: We assessed the changes of bone mass in patients with hyperthyroidism by measuring bone mineral density using a new method, dual energy X-ray absorptiometry. DESIGN: The values of bone mineral density in patients with hyperthyroidism were compared with data obtained from the controls, and we assessed the correlation analysis between bone mineral density and several metabolic parameters. PATIENTS: We studied 52 Japanese patients with hyperthyroidism (20 males, 32 females). Healthy normal subjects served to establish the mean bone mineral density in the healthy Japanese population (Shiraki et al. 1991). MEASUREMENT: Bone mineral density was assessed by the measurement of lumbar vertebrae and femur by dual energy X-ray absorptiometry. The bone mineral density of vertebrae for each patient was calculated as the percentage of the mean value (% bone mineral density) obtained from an age and sex-matched control group. Blood was drawn to measure the levels of serum calcium, phosphorus, creatinine, alkaline phosphatase, free T3, free T4, TSH, TSH receptor antibody, parathyroid hormone, and serum osteocalcin. RESULTS: The percentage bone mineral density of vertebrae in patients was 92.6 as compared with that of normal controls, and was inversely correlated with serum TSH receptor antibody, osteocalcin, and alkaline phosphatase. CONCLUSIONS: These findings suggest that bone mineral density is decreased in patients with hyperthyroidism and that TSH receptor antibody, osteocalcin, and alkaline phosphatase are sensitive markers of bone metabolism alterations in hyperthyroidism.

Bone mineral density measured by dual energy x-ray absorptiometry in patients with non-insulin-dependent diabetes mellitus.

Because of the previous controversial findings in diabetic patients with older methodologies, we assessed bone mineral density (BMD) in 78 patients (38 males and 40 females) with non-insulin-dependent diabetes mellitus using dual energy x-ray absorptiometry (DEXA). BMD was measured in lumbar vertebrae (L2-4). The BMD of each patient was calculated as the percentage of the mean value (%BMD) obtained from a healthy control group matched for sex and age. The %BMD of the patients with diabetes was about 100% for females and 96% for males, as compared with BMD of normal controls. The %BMD of the patients with diabetes was significantly correlated with body mass index and urinary C peptide level, and inversely correlated with age and duration of diabetes within 20 years. No relationships were found between %BMD and serum calcium, phosphorus, or glycosylated hemoglobin A1C levels. These observations suggest that metabolic abnormalities associated with diabetes mellitus alter the BMD, and that such factors as duration of the disease and deficit in insulin secretion are risk factors for decreased BMD.