Molecular cloning of type I collagen cDNA and nutritional regulation of type I collagen mRNA expression in grass carp.

Grass carp (Ctenopharyngodon idellus) are important Chinese freshwater fish, and in China, the faba bean has been used as the sole food source for grass carp to transform them into crisp grass carp. Because of this, crisp grass carp has become an economically important fish because of its increased muscle hardness. To study the nutritional regulation of type I collagen in faba bean-fed grass carp, we isolated type I collagen alpha 2 (COL1A2) on the basis of our isolation of COL1A1. The COL1A2 cDNA was found to be 4899 bp in length and included a 4059-bp coding sequence (CDS) and encoded a polypeptide of 1352 AA. The protein peptide molecular weight was 127.39 kD, and the theoretical isoelectric point was 9.37. The COL1A2 protein possessed five α-helixes, eight ß-sheets, 16 regions of triple helical repeats, 21 low-complexity regions, 10 function domains and two zinc-binding sites; however, no calcium-binding sites were observed. The mRNA expression of COL1A1 and COL1A2 was assessed in eight tissues (muscle, hepatopancreas, intestine, gills, skin, fin, kidney and spleen) from grass carp and crisp grass carp by semi-quantitative RT-PCR. Expression of COL1A1 in the muscle, intestines and skin of crisp grass carp was higher than that in grass carp, and expression of COL1A2 in the muscle, gills, fin and skin of crisp grass carp was higher than that in grass carp. In the muscle of crisp grass carp, expression of COL1A1 and COL1A2 was higher than that in grass carp, which was further confirmed by real-time PCR, and collagen content also was enhanced. These results demonstrated that type I collagen was closely related to the increased muscle hardness of faba bean-fed grass carp.

Electrospun biphasic drug release polyvinylpyrrolidone/ethyl cellulose core/sheath nanofibers.

The capability of core/sheath nanofibers prepared using coaxial electrospinning to provide adjustable biphasic drug release was investigated. Using ketoprofen (KET) as the model drug, polyvinylpyrrolidone as the sheath polymer, and ethyl cellulose as the core matrix, the coaxial process could be conducted smoothly and continuously without spinneret clogging. Scanning electron microscopy and transmission electron microscopy revealed linear nanofibers with homogeneous and clear core/sheath structures. Differential scanning calorimetry and X-ray diffraction verified that the core/sheath nanofibers were nanocomposites, with the drug present in the polymer matrix in an amorphous state. Attenuated total reflectance-Fourier transform infrared spectra demonstrated that the sheath polymer and core matrix were compatible with KET owing to hydrogen bonding. In vitro dissolution tests showed that the core/sheath nanofibers could provide typical biphasic drug release profiles consisting of an immediate and sustained release. The amount of drug released in the first phase was tailored by adjusting the sheath flow rate, and the remaining drug released in the second phase was controlled by a typical diffusion mechanism. The present study shows a simple and useful approach for the systematic design and fabrication of novel biomaterials with structural characteristics for providing complicated and programmed drug release profiles using coaxial electrospinning.

Total hip replacement for high dislocated hips without femoral shortening osteotomy.

When performing total hip replacement (THR) in high dislocated hips, the presence of soft-tissue contractures means that most surgeons prefer to use a femoral shortening osteotomy in order to avoid the risk of neurovascular damage. However, this technique will sacrifice femoral length and reduce the extent of any leg-length equalisation. We report our experience of 74 THRs performed between 2000 and 2008 in 65 patients with a high dislocated hip without a femoral shortening osteotomy. The mean age of the patients was 55 years (46 to 72) and the mean follow-up was 42 months (12 to 78). All implants were cementless except for one resurfacing hip implant. We attempted to place the acetabular component in the anatomical position in each hip. The mean Harris hip score improved from 53 points (34 to 74) pre-operatively to 86 points (78 to 95) at final follow-up. The mean radiologically determined leg lengthening was 42 mm (30 to 66), and the mean leg-length discrepancy decreased from 36 mm (5 to 56) pre-operatively to 8.5 mm (0 to 18) postoperatively. Although there were four (5%) post-operative femoral nerve palsies, three had fully resolved by six months after the operation. No loosening of the implant was observed, and no dislocations or infections were encountered. Total hip replacement without a femoral shortening osteotomy proved to be a safe and effective surgical treatment for high dislocated hips.

Effects of erbium,chromium:YSGG laser irradiation on canine mandibular bone.

BACKGROUND: Only relatively few reports have described the morphological effects on bone produced by erbium,chromium: yttrium,scandium,gallium,garnet (Er,Cr:YSGG) laser irradiation, and none has investigated the atomic changes or estimated the temperature increases involved. The objectives of this study were to investigate the morphological, atomic, and temperature changes in irradiated areas during and after laser irradiation, and to evaluate the cutting effect on canine mandibular bone in vitro. METHODS: Two canine mandibular bones were cut into 3 to 5 cm pieces and irradiated by an Er,Cr:YSGG laser utilizing a water-air spray at 5 W and 8 Hz for 10 or 30 seconds. During and after laser irradiation, temperature increases in the irradiated areas were measured by thermography. The samples were then observed by stereoscopy and scanning electron microscopy to determine morphological changes and by energy dispersive x-ray spectroscopy to evaluate atomic alterations. RESULTS: Regular holes or grooves having sharp edges and smooth walls were produced, but no melting or carbonization was observed. The maximum temperature increase was an average 12.6 degrees C for 30-second irradiation. The continuous time of a temperature increase of more than 10 degrees C was consistently less than 10 seconds. An atomic analytical examination revealed that the calcium:phosphorus ratio was not significantly changed between the lased and unlased areas (P>0.0 1). CONCLUSION: These results showed that the Er,Cr:YSGG laser cuts canine mandibular bone effectively without burning, melting, or altering the calcium:phosphorus ratio of the irradiated bone.

Effects of erbium,chromium:YSGG laser irradiation on root canal walls: a scanning electron microscopic and thermographic study.

The present study evaluates the morphological changes in root canal walls and temperature changes at root surfaces as a result of intracanal irradiation by erbium,chromium:YSGG laser under various conditions in vitro. Sixty single-rooted human teeth were examined. Root canals were prepared, and laser irradiation was performed using an optic fiber at output powers ranging from 1 to 6 W with or without water spray cooling. Specimens were evaluated by stereoscopy, scanning electron microscopy, and thermography. Carbonization and cracks were observed in all samples irradiated without cooling, whereas little or no carbonization and no smear layer or debris were observed in samples irradiated with cooling. Maximum temperature rise at irradiation without cooling was above 37 degrees C, whereas that at irradiation with cooling was 8 degrees C. Results of the present study indicate that erbium,chromium:YSGG laser irradiation with water spray cooling is a useful method for removal of smear layer and debris from root canals.

Effects of erbium, chromium:YSGG laser irradiation on root surface: morphological and atomic analytical studies.

OBJECTIVE: The purpose of this study was to investigate the morphological and atomic changes on the root surface by stereoscopy, field emission-scanning electron microscopy (FE-SEM), and energy dispersive X-ray spectroscopy (SEM-EDX) after erbium, chromium:yttrium, scandium, gallium, garnet (Er,Cr:YSGG) laser irradiation in vitro. SUMMARY BACKGROUND DATA: There have been few reports on morphological and atomic analytical study on root surface by Er,Cr:YSGG laser irradiation. METHODS: Eighteen extracted human premolar and molar teeth were irradiated on root surfaces at a vertical position with water-air spray by an Er,Cr:YSGG laser at the parameter of 5.0 W and 20 Hz for 5 sec while moving. The samples were then morphologically observed by stereoscopy and FE-SEM and examined atomic-analytically by SEM-EDX. RESULTS: Craters having rough but clean surfaces and no melting or carbonization were observed in the samples. An atomic analytical examination showed that the calcium ratio to phosphorus showed no significant changes between the control and irradiated areas (p > 0.01). CONCLUSIONS: These results showed that the Er,Cr:YSGG laser has a good cutting effect on root surface and causes no burning or melting after laser irradiation.

Study on acid resistance of human dental enamel and dentin irradiated by semiconductor laser with Ag(NH3)2F solution.

OBJECTIVE: The purpose of this study was to investigate the acquired acid resistance of human dental enamel and dentin treated by 38% diamine silver fluoride [Ag(NH3)2F] solution and semiconductor (diode) laser irradiation in vitro. BACKGROUND DATA: There have been no reports on the acid-resistant effect by combined use of fluoride and semiconductor laser. METHODS: Sixty crowns of extracted human molars were divided into two groups for enamel and dentin samples, and each group was subdivided into three subgroups of 10 each. Each subgroup of enamel and dentin samples served as a control; one was treated with Ag(NH3)2F and the other was treated with Ag(NH3)2F and semiconductor laser irradiation at 2 W for 30 sec. Then all samples were immersed in 5 ml of 0.1 M lactic acid (pH 4.8) at 37 degrees C for 24 h. The concentration of calcium ion (Ca2+) dissolved in the solution was measured by atomic absorption spectrophotometry, and the samples were observed by stereoscopy and scanning electron microscopy (SEM). RESULTS: In both enamel and dentin samples, dissolved Ca2+ concentration in Ag(NH3)2F- or Ag(NH3)2F- and laser-treated group was significantly decreased compared with that in the control (p < 0.01). SEM findings showed that numerous cubic particles ranging from 0.1 to 0.5 mcirom were observed only in the combined treated groups of both enamel and dentin samples. CONCLUSIONS: A significantly decreased solubility of human enamel and dentin was acquired after treatment by Ag(NH3)2F and semiconductor laser irradiation, which suggested that this combined use has the capability of a more efficient acid-resistant effect on human dental hard tissues.

Study on removal effects of filling materials and broken files from root canals using pulsed Nd:YAG laser.

OBJECTIVES: The aims of this study were to investigate the capability of removing the filling materials or broken files from root canals with pulsed Nd:YAG laser irradiation at three parameters, and to evaluate the temperature rises on root surfaces and morphological changes of root canal walls in vitro. SUMMARY BACKGROUND DATA: There has been no report on removing the filling materials and broken instruments from root canals with laser irradiation. MATERIALS AND METHODS: Thirty-six extracted human incisors were divided into two groups of 18 each. Eighteen root canals were shaped and obturated with gutta-percha and root canal sealer. In another 18 specimens, 3-mm long fractures of files were performed intentionally during root canal preparation. A pulsed Nd:YAG laser irradiation was performed repeatedly at three parameters of 1, 2, and 3 W to remove the filling materials or file fragments. Temperature rises on root surfaces during laser irradiation were measured by thermography, and the remaining of obturated materials and morphological changes of root canal walls were evaluated by stereoscopy and scanning electron microscopy. RESULTS: In more than 70% of the teeth, the obturated materials were completely removed by laser, and in more than 55% of the teeth, the broken files were successfully removed. Temperature rises ranging from 17 degrees C to 27 degrees C were measured from 6 to 11 repeated times. CONCLUSION: These results demonstrated that a pulsed Nd:YAG laser irradiation has the capability of removing the obturated materials in root canals and is useful for removing the broken files in same if the counter-measure reducing the temperature rise is performed.

Morphological and atomic analytical studies on enamel and dentin irradiated by an erbium, chromium:YSGG laser.

OBJECTIVE: The purposes of this study were to investigate the morphological and atomic analytical changes and to evaluate the cutting effect on dental hard tissues of this laser in vitro. BACKGROUND DATA: There have been few reports on morphological and atomic analytical study of dental hard tissues after erbium,chromium:yttrium,scandium,gallium,garnet (Er,Cr:YSGG) laser irradiation. METHODS: Eighteen extracted human molars were sectioned into 3-mm-thick slices, which were irradiated with water-air spray by an Er,Cr:YSGG laser at 6.0 W and 20 Hz for 5 sec for enamel and 5.0 W and 20 Hz for 5 sec for dentin. The samples were then morphologically observed and examined atomic-analytically. RESULTS: Regular holes having sharp edges and smooth walls, but no melting or carbonization, were observed in both samples. An atomic analytical examination showed that the calcium ratio to phosphorus showed no significant changes between the lased areas and unlased areas (p > 0.01). CONCLUSION: These results showed that the Er,Cr:YSGG laser has a good cutting effect on dental hard tissues and offers advantages of no burning or melting after laser irradiation.

[Isoelectric focusing analysis of Echinococcus multilocularis protoscolex antigens].

OBJECTIVE: To understand the components of the crude antigens of Echinococcus multilocularis protoscoleces. METHODS: Isoelectric focusing was carried out to fractionate the protein components of E. multilocularis protoscoleces. The individual fractions were collected and subjected to SDS-PAGE, the gels were analyzed by a gel analysis system. RESULTS: The pI values of individual fractions varied from 4.0 to 8.5, while the molecule weights of main protein bands were 72/73, 62/63, 42/44, 35/36, 34 and 21/22 kDa. CONCLUSION: An alternative separation method of crude antigens of Echinococcus multilocularis was established.

Alveolar echinococcosis: Em2plus-ELISA and Em18-western blots for follow-up after treatment with albendazole.

Eleven cases of alveolar echinococcosis (Echinococcus multilocularis infection) with non-resectable lesions but treated with albendazole for 17 to 69 months were followed-up clinically and serologically for 4.5-11.5 years. Based on the clinical outcome and computerized tomography (CT) scanning, they were divided into 4 groups of 2 cured cases, 5 stabilized cases, 3 cases with recurrences, and one treatment failure. Forty-seven sequentially collected sera from the 11 cases were analysed by sequential enzyme-linked immunosorbent assay (ELISA) using Em2plus antigen (Em2plus-ELISA) and Western blotting to detect antibody response against Em18 (Em18-Western blots). The antibody levels in one of the cured and 2 of the stabilized cases fell below the cut-off level in the Em2plus-ELISA 4.5-6 years after effective treatment, whereas all other cases, including 2 of those with recurrences, showed large reductions initially but increased again during the follow-up period. Em18-Western blots of the 2 cured cases and 2 of the stabilized cases became negative. IgG subclasses with responses against Em18 which fell to zero included IgG1 (2), IgG3 (one) and IgG4 (one). All other cases showed no decrease in antibody response against Em18. There were, in general, reasonably reliable correlations between the success or failure of chemotherapy and antibody responses by Em2plus-ELISA and Em18-Western blots. These results suggest that both Em2plus-ELISA and Em18-Western blot are potentially useful in evaluating and predicting the efficacy of chemotherapy.

The long-term effects of aerosol pentamidine on pulmonary function. The Toronto Aerosolized Pentamidine Study (TAPS) Group.

Aerosolized pentamidine (AP) has been widely used for prophylaxis of pneumocystis carinii pneumonia (PCP) since 1988. The objective of this study was to evaluate the long-term effects of AP on pulmonary function. Of 36 patients with AIDS who were receiving AP for secondary prophylaxis of PCP, 13 patients had been using AP continuously for more than 52 weeks. AP was given using a Fisoneb ultrasonic nebulizer with five loading doses of 60 mg over two weeks, followed by one dose of 60 mg every two weeks. Baseline PFT were TLC 92 +/- 14% pred, FVC 90 +/- 11% pred, FEV1 91 +/- 11% pred, FEF25-75 95 +/- 17% pred, and DLCO (corrected for hemoglobin) 70 +/- 22% pred. No significant change in TLC, FVC, FEV1, or DLCO was seen after 56 weeks of AP. There was a 20% fall in FEF25-75 seen after 56 weeks, which was statistically significant. However, the clinical significance of a fall of this magnitude in the FEF25-75 is uncertain. Similar results were seen in a smaller subset of patients who received AP for at least 76 weeks. Although the small sample size must be considered, this data suggests that there is no clinically significant change in pulmonary function associated with the use of AP for up to 76 weeks.

Respiratory muscle weakness in Charcot-Marie-Tooth disease. A field study.

A field study on respiratory muscle function and basic pulmonary mechanics was conducted at an international convention of 200 individuals with Charcot-Marie-Tooth disease. Forty patients with respiratory symptoms were evaluated; 15 had significant respiratory muscle dysfunction and 25 were normal. The results suggest that proximal upper limb involvement in individuals with Charcot-Marie-Tooth disease is a simple but significant predictor of respiratory muscle dysfunction.