[Species Identification Based on Morphological Variability of Femur].

OBJECTIVE: To study the morphological characteristics of femurs of adult human and 11 kinds of adult animals from cattle, horses, pigs, goats, sheep, dogs, cats, rabbits, geese, ducks, chickens, and to establish an effective species identification method among various species. METHODS: The 4 cm mid-diaphyseal segment of the femur from adult human (older than 20 years old) at autopsy was obtained. Addi-tionally, the 4 cm ones from 11 kinds of adult animals were obtained. After decalcification, all femurs were made into slices, and then were observed by optical microscope. The 25 indexes were selected and analyzed by step discriminant analysis according to differences between human and mammal, human and poultry, and human and 11 kinds of animals. RESULTS: The histological structure of bone mineral density of middle part of femur had obvious characteristics among the species. And the morphology and number of osteon showed the trend of obvious biological evolution. There were 11 indexes with significant differences between human and 11 kinds of animals to establish some mathematical models to discriminate all species. The correct discrimination rate was 96.3% between human and mammal. The correct discrimination rate was up to 100% between human and poultry, and was 89.4% among human, mammal and poultry. CONCLUSION: The mathematical models have good correct discrimination rate among human and the other animals, which could be applied in the practical species identification cases.

[Influences of cycle index and volume on sensitivity of DNA detection].

OBJECTIVE: To explore the feasibility of improving the sensitivity of DNA detection by increasing the PCR cycle index and decreasing the volume of amplifying system. METHODS: The DNA of semen were collected from 10 healthy irrelevant volunteers, and were quantified to 50, 40, 30, 25, 20, 15, 10 pg/microL, separately. All samples were then amplified in 10, 5, 3 microL volume and at 28, 30, 32, 34, 36 cycles, respectively. 3130 genetic analyzer was used to detect 15 autosomal STR loci. RESULTS: Under the situation of 28 cycles and 3 microL volume, samples which achieved > 40 pg/microL could be correctly typed. Under the situation of 10, 5, 3 microL volume, samples which achieved > 20 pg/microL could be correctly typed at 34 cycles. When increasing the index to 36 cycles, they could not be correctly typed because of the non-specific band. CONCLUSION: DNA detecting sensitivity can be improved to a certain extent by increasing the cycle index and decreasing the volume of amplifying system.

Value of radiographic esophageal imaging in determining an optimal atrial septal puncture site for percutaneous balloon mitral valvuloplasty.

OBJECTIVE: To investigate the value of radiographic esophageal imaging in facilitating transseptal catheterization in patients undergoing percutaneous balloon mitral valvuloplasty. SUBJECTS AND METHODS: A total of 468 patients were randomized into either the study group (n = 234), in which radiographic esophageal imaging by the oral administration of a contrast media took place, or the control group (n = 234), in which the Ross technique was used. Of the 468 patients, 203 were males and 265 were females. The average ages of the study and control groups were 53 +/- 16 and 51 +/- 17 years, respectively. The patients had severe left atrial enlargement, as measured using 2-dimensional echocardiography. RESULTS: In the study group, the left atrial impression on the esophagus was clearly seen, and was used to identify the puncture site on the right atrial side for the passage of the transseptal catheter. In the control group, the left atrial silhouette was not clearly shown by fluoroscopy in 112 patients (47.5%). The success rate of transseptal catheterization in the study group was higher than in the control group (99.6 vs. 45.7%, p = 0.0001). There were no complications in the study group, but pericardial tamponade occurred in 1 patient in the control group. CONCLUSION: Radiographic esophageal imaging facilitates the identification of an optimal atrial transseptal puncture site, and improves the success rate of transseptal catheterization in patients undergoing percutaneous balloon mitral valvuloplasty.

Effects of folic acid on cardiac myocyte apoptosis in rats with streptozotocin-induced diabetes mellitus.

BACKGROUND: The effect of folic acid on cardiac myocyte apoptosis secondary to diabetes is unknown. METHODS: Diabetic rats were divided into diabetic control (DC, n = 11), low-dose (LDF, 0.4 mg/kg/day, n = 12) and high-dose (HDF, 1.2 mg/kg/day, n = 12) folic acid groups. Non-diabetic rats (n = 11) were used as the normal control (NC). RESULTS: After 11 weeks of treatment, compared with the NC group, the DC group showed a reduced blood levels of reactive oxygen species (ROS, P < 0.01). The rate of cardiac myocyte apoptosis in the diabetic control group was also greater than in the non-diabetic control group (P < 0.01). In folic acid-treated rats, the blood levels of ROS was higher than in the diabetic control group (P < 0.05). There was a dose-dependent reduction in the rate of cardiac myocyte apoptosis in the folic acid groups (P < 0.01), and this was accompanied by an increased level of anti-apoptotic protein Bcl-2 and decreased level of pro-apoptotic protein Bax and Fas (P < 0.01). CONCLUSIONS: Dietary folic acid supplementation diminishes the cardiac myocyte apoptosis in streptozotocin-induced diabetes. The apoptosis suppression is accompanied by an increase in the expression of Bcl-2 and a decrease in Bax and Fas.

[Comparative research of the influence factors of DNA extraction of bloodstain on the filter paper with Chelex-100 method].

OBJECTIVE: To improve DNA extraction from bloodstain on the filter paper and to establish a rapid, simple, and cost-effective method for DNA extraction suitable for database construction. METHODS: Seven hundred and fifty two aged bloodstains on filter paper were randomly divided into four groups. The four different DNA extraction methods were compared with each other, and two DNA extraction methods used for 63 fresh bloodstains on filter paper were also compared with each other. RESULTS: There were no statistically significant differences observed among the four DNA extraction methods (P > 0.05) for aged bloodstains on filter paper; But the difference between the two DNA extraction methods for fresh bloodstains on filter paper was obviously (P < 0.05). CONCLUSION: Extraction of DNA samples from aged bloodstains on filter paper can be accomplished by using Chlex-100 methodology directly with no need to wash the bloodstains.

[Effect of interleukin-6 on gene expression of certain cytokines during wound healing process of mouse skin].

To examine the effect of the interlukin-6 (IL-6) in wound healing process, gene expression profiles of cytokines including interlukin-1alpha (IL-1alpha), interlukin-1beta (IL-1beta), keratinocyte chemoattractant (KC), macrophage inflammatory protein-1alpha (MIP-1alpha), and macrophage inflammatory protein-2 (MIP-2) during the skin wound healing on the IL-6(+/+) and IL-6(-/-) mice were detected using immunochemical staining and RT-PCR methods at different phases after wound. The results showed that these cytokines were expressed at early phage after wound, and a expressing peak were found on the third day after wound, and decreased on the sixth day after wound. Meanwhile, on the third and sixth day after wound, all the levels of the five cytokines expressed of the IL6(-/-) mice were significantly lower than those of the IL-6(+/+) mice, but on the first day after wound, only the levels of MIP-1alpha and KC of the IL-6(-/-) mice were lower. The process of skin wound healing was later in IL-6(-/-) mice than that in IL-6(+/+) mice, but it was complete in IL-6(-/-) mice. These results showed that IL-6 induce expression of the other five cytokines detected, and advance the repair of the wound on skin of mice, but in the mice of IL-6 deletion, the wound healing process was not disturbed significantly.

Essential involvement of IL-6 in the skin wound-healing process as evidenced by delayed wound healing in IL-6-deficient mice.

To clarify interleukin (IL)-6 roles in wound healing, we prepared skin excisions in wild-type (WT) and IL-6-deficient BALB/c [knockout (KO)] mice. In WT mice, the wound area was reduced to 50% of original size at 6 days after injury. Microscopically, leukocyte infiltration was evident at wound sites. Furthermore, the re-epithelialization rate was approximately 80% at 6 days after injury with increases in angiogenesis and hydroxyproline contents. The gene expression of IL-1, chemokines, adhesion molecules, transforming growth factor-beta1, and vascular endothelial growth factor was enhanced at the wound sites. In contrast, the enhanced expression of these genes was significantly reduced in KO mice. Moreover, in KO mice, the reduction of wound area was delayed with attenuated leukocyte infiltration, re-epithelialization, angiogenesis, and collagen accumulation. Finally, the administration of a neutralizing anti-IL-6 monoclonal antibody significantly delayed wound closure in WT mice. These observations suggest that IL-6 has crucial roles in wound healing, probably by regulating leukocyte infiltration, angiogenesis, and collagen accumulation.