HNA-3a and HNA-3b antigens among 9 ethnic populations and the Han population in Southwest China.

BACKGROUND: Human neutrophil antigen 3 (HNA-3) is encoded by the SLC44A2 gene. Antibodies against HNAs can cause severe, often fatal, transfusion reactions, known as transfusion-related acute lung injury, and neonatal neutropenia. We explored the 2 common HNA-3 variants in 9 ethnic populations residing in Sichuan and Yunnan provinces of China as compared to the Han population. METHODS: We genotyped for SLC44A2 (rs2288904) by polymerase chain reaction sequence-based typing among blood donors, for a total of 2206 individuals in Yunnan and 376 in Sichuan. RESULTS: The SLC44A2*02 allele (HNA-3b antigen) frequency varied between 0.24 and 0.33 for all 9 ethnic populations in Yunnan, including Zhuang, Derung, Hani, Lisu, Bai, Miao, Dai, Naxi, and Yi. Specifically, the Yi ethnicity did not present an unusually great SLC44A2*02 frequency at any of the 4 locations examined in Yunnan. Except of the Yi ethnicity in Sichuan (0.40), the Han ethnicity, as the majority population group, had the greatest SLC44A2*02 frequency with 0.39 in Yunnan and 0.35 in Sichuan. CONCLUSION: The ethnic populations in Southwest China are not at an increased risk for anti-HNA3a compared to the Han population, with the possible exception of Yi in Sichuan. Our data, however, corroborated the known high prevalence of SLC44A2*02 in Han populations. Hence, the Han populations in Yunnan, Sichuan and elsewhere in China are at a comparatively great risk for developing HNA-3a antibodies.

Antiproliferative effect of echinacoside on rat pulmonary artery smooth muscle cells under hypoxia.

The main purpose of this study is to evaluate the effect of echinacoside (ECH) on hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells (PASMCs) and the underlying mechanism. PASMCs were incubated under normoxia (nor), hypoxia (hyp), hypoxia + 0.35 mM ECH (hyp + ECH0.35), or hypoxia + 0.4 mM ECH (hyp + ECH0.4) for 24 h. Cell viability was assessed by MTS assays. The morphology of apoptosis was observed by DAPI staining, and apoptosis was quantified by flow cytometric analysis. Caspase-3 activity was determined by immunohistochemistry and real-time PCR, and the expressions of HIF-1α, Bax, Bcl-2, and Fas were determined by real-time PCR. Hypoxia induced significant proliferation of PASMCs, which could be inhibited by ECH in a concentration-dependent manner. This was associated with apoptosis of PASMCs. Z-DEVD-FMK could partly reduce the suppression effect of ECH; protein and gene expression of caspase-3 were significantly higher in the hyp + ECH0.4 and hyp + ECH0.35 groups. ECH significantly increased the expressions of Bax and Fas, but decreased the expressions of Bcl-2 and HIF-1α. ECH could inhibit hypoxia-induced proliferation of rat PASMCs, which is associated with apoptosis of PASMCs and improvement of hypoxia. ECH might be a potential agent for prevention and treatment of hypoxia-induced PAH.

Advances of regenerated and functionalized silk biomaterials and application in skin wound healing.

The cocoon silk of silkworms (Bombyx mori) has multiple potential applications in biomedicine due to its good biocompatibility, mechanical properties, degradability, and plasticity. Numerous studies have confirmed that silk material dressings are more effective than traditional ones in the skin wound healing process. Silk material research has recently moved toward functionalized biomaterials and achieved remarkable results. Herein, we summarize the recent advances in functionalized silk materials and their efficacy in skin wound healing. In particular, transgenic technology has realized the specific expression of human growth factors in the silk glands of the silkworms, which lays the foundation for fabricating novel and low-cost functionalized materials. Without a green and safe preparation process, the best raw silk materials cannot be made into medically safe products. Therefore, we provide an overview of green and gentle approaches for silk degumming and silk sericin (SS) extraction. Moreover, we summarize and discuss the processing methods of silk fibroin (SF) and SS materials and their potential applications, such as burns, diabetic wounds, and other wounds. This review aims to enhance our understanding of new advances and directions in silk materials and guide future biomedical research.

[Health Risk Assessment and Priority Control Factors Analysis of Heavy Metals in Agricultural Soils Based on Source-oriented].

To analyze the source apportionment and health risk of heavy metals in agricultural soils of major producing areas of agricultural products in Chongqing, a positive matrix factorization (PMF) model and health risk assessment (HRA) model based on Monte Carlo simulation were used. Meanwhile, both the PMF and HRA model were combined to explore health risks of heavy metals in agricultural soils by different pollution sources in order to determine the priority control factors. The results showed that the average values of Cd concentration were higher than its corresponding background value; the average values of Cr concentration were lower than its corresponding background value; and the average values of As, Pb, Cu, Ni, and Zn concentration were basically consistent with their corresponding background values. Using PMF model analysis, natural sources, industrial sources, and agricultural sources were identified as the determinants for the accumulation of heavy metals in agricultural soils, with the contribution rates of 35%, 24%, and 41%, respectively. Using the HRA model based on Monte Carlo simulation analysis, carcinogenic risks of adult and children were tolerable (1.00E-6 < TCR ≤ 1.00E-4), whereas non-carcinogenic risks were acceptable (HI ≤ 1). Oral ingestion was the main exposure pathway. The analysis results of the relationship among heavy metals, pollution sources, and health risks showed that industrial pollution and As were identified as priority control factors, and agricultural pollution and Cd were identified as secondary control factors. Our findings provide scientific support for decision makers to control soil pollution and reduce the management costs of soil pollution.

Silk Sericin and Its Composite Materials with Antibacterial Properties to Enhance Wound Healing: A Review.

Wound infections may disrupt the normal wound-healing process. Large amounts of antibiotics are frequently used to prevent pathogenic infections; however, this can lead to resistance development. Biomaterials possessing antimicrobial properties have promising applications for reducing antibiotic usage and promoting wound healing. Silk sericin (SS) has been increasingly explored for skin wound healing applications owing to its excellent biocompatibility and antioxidant, antimicrobial, and ultraviolet-resistant properties. In recent years, SS-based composite biomaterials with a broader antimicrobial spectrum have been extensively investigated and demonstrated favorable efficacy in promoting wound healing. This review summarizes various antimicrobial agents, including metal nanoparticles, natural extracts, and antibiotics, that have been incorporated into SS composites for wound healing and elucidates their mechanisms of action. It has been revealed that SS-based biomaterials can achieve sustained antimicrobial activity by slow-release-loaded antimicrobial agents. The antimicrobial-loaded SS composites may promote wound healing through anti-infection, anti-inflammation, hemostasis, angiogenesis, and collagen deposition. The manufacturing methods, benefits, and limitations of antimicrobial-loaded SS materials are briefly discussed. This review aims to enhance the understanding of new advances and directions in SS-based antimicrobial composites and guide future biomedical research.

[Health Risk Assessment and Environmental Benchmark of Cadmium in Farmland Soils around the Gangue Heap of Coal Mine, Chongqing].

To analyze the health risk assessment and environmental benchmark of cadmium in farmland soils surrounding the gangue heap of a coal mine in Chongqing, Hakanson, the ecological risk index and health risk assessment were used. Meanwhile, the soil environmental reference value of the regional cultivated land was inverted based on the species sensitive distribution model (SSD). The results showed that the dryland soil was polluted by Cd, with an over-standard rate of 55.8%, and the paddy field soil was polluted by Cd, with an over-standard rate of 31.6%. The corn was polluted by Cd, with an over-standard rate of 4.4%, and the rice was not polluted by Cd. The Hakanson ecological risk index showed that Cd was mainly characterized in soils by high ecological risk and considerable ecological risk. The health risk assessment indicated that Cd presented low non-carcinogenic risk by corn and rice; however, it showed acceptable carcinogenic risk by corn and unacceptable carcinogenic risk by rice in this study. The sensitivity analysis of health risks showed that the content of Cd was the most sensitive. The SSD inversion showed that the reference values for Cd in dryland soil of pH ≤ 5.5, 5.57.5 had HC5 values of 0.491, 0.382, 0.376, and 0.588 mg·kg-1, respectively, and that for Cd in paddy soil had an HC5 value of 0.807 mg·kg-1. The reverse analysis showed that the HC5 of Cd in dryland soil (pH ≤ 7.5) and paddy soil was relatively relaxed and was higher than the soil risk screening values, which showed that the current standard was relatively loose. However, the HC5 of Cd in dryland soil (pH>7.5) was lower than the soil risk screening values, which showed that the current standard was relatively strict. It is suggested that the current soil standard could be adjusted in this area.

[Health Risk Assessment of Heavy Metals in Agricultural Soils Around the Gangue Heap of Coal Mine Based on Monte Carlo Simulation].

To analyze the pollution characteristics of heavy metals in the soil of farmland surrounding the gangue heap of a coal mine in Chongqing, the Nemerow, Muller, and Hakanson indices were used. Meanwhile, to investigate the health risks of heavy metals in soil, a health risk assessment model was employed using Monte Carlo simulation. The results revealed that the average contents of Cd, Hg, As, Pb, Cr, Cu, Ni, and Zn were higher than their soil background values, and the Muller index (Igeo) from high to low was as follows:Cd>Hg>Cu>As>Ni>Zn>Cr>Pb. The Nemerow index (PN) demonstrated that the results of the assessment indicated mainly light pollution (1Cu>Ni>Cr>Zn>As>Pb>Hg. The Hakanson index (RI) demonstrated that the results of the assessment indicated mainly moderate ecological risk (150 ≤ RI<300). The single ecological risk index (Eri) from high to low was as follows:Cd>Hg>As>Cu>Ni>Pb>Cr>Zn. The health risk assessment of heavy metals revealed that the non-carcinogenic health risks of children and adults could be ignored, but carcinogenic health risks existed, with an average total carcinogenic risk index (TCR) of 1.04E-5 and 3.94E-6, respectively. The major carcinogenic factors were As and Cd. Sensitivity analysis of non-carcinogenic health risks revealed that the ingestion rate of soil (Ringest) was the most sensitive, followed by the contents of As. Meanwhile, sensitivity analysis of carcinogenic health risks indicated that the content of As was the most sensitive, followed by the ingestion rate of soil (Ringest). In general, the health risks of children, including non-carcinogenic and carcinogenic risks, were higher than those of adults.

[Expression changes of BDNF in lung of rats subjected to brain ischemia].

OBJECTIVE: To investigate the expression of brain-derived neurotrophic factor (BDNF) in lung injury induced by brain ischemia in rats. METHODS: 46 adult SD rats were assigned randomly to sham operation group and brain ischemia lung injury group (BILI, n = 23 in each group). Rats were subjected brain ischemia and allowed to survived 3 d. After performed neurological functional severe deficit evaluation, lung edema was observed (n=5). The BDNF expression for its mRNA and protein in lung tissues was determined by using ELISA (n=5) and RT-PCR technique (n=8). The localization of BDNF was also determined by immunohistochemistry (n=5). RESULTS: After brain ischemia for 3 days, the severe neurological functional deficit and edema in lung were seen. BDNF was located in cytoplasma of smooth muscle and epithial cells in the lung. The level of BDNF mRNA (indicated by RT-PCR) and the protein level (indicated by ELISA) were all upregulated at 3 days after brain ischemia (P<0.05). CONCLUSION: Lung edema occurred after brain ischemia in rats is concomitant with BDNF expression, which consists of the mechanism involved lung injury induced by brain ischemia.

[Expression changes of MEK in lung of rats subjected to brain ischemia].

OBJECTIVE: To explore the expression changes of mitogen extracellular kinase (MEK) in injured lung after brain ischemia in rats. METHODS: Adult SD rats were assigned randomly to sham operation group and brain ischemia lung injury (BILI) group. Rats in BILI group were subjected brain ischemia and allowed to survived 3 d. Pathalogical changes in lung were indicated by HE staining. The MEK expression was determined by RT-PCR and Western blot technique. RESULTS: After brain ischemia, the bulk of inflammatory cells invaded into lung were observed. Upregulated level of MEK mRNA and protein were found at 3 days after ischemia (P<0.05). CONCLUSION: The upregulated expression of MEK implied that the MEK may play some roles in lung injury after brain ischemia.

[Constrution of human single herpes virus carried BDNF gene vector and its bioactivity evaluation].

OBJECTIVE: To construct the recombinant vector of human single herpes virus (HSV) carried brain derived neurotrophic factor (BDNF) gene. METHODS: BDNF gene was acquired from rat brain tissue by RT-PCR, then was cloned into plasmid, and enveloped by HSV. The recombinant was used to transfer cultured cortical neurons. The number and neurite length of neurons were quantified. The BDNF level and subcellular localization were detected by ELISA and immunohistochemistry. RESULTS: HSV carried BDNF gene recombinant has been successfully constructed. The recombinant showed the bioactivity on the growth of cortical neurons. BDNF level was increased significantly in BDNF transferred group. CONCLUSION: HSV carried BDNF gene recombinant, with the bioactivity, has been successfully constructed. This could provide the vector for the treatment of BDNF under disease condition base on transferring gene technique.

[Effect of moxibustion on colonic mucosal injury and expression of CGRP positive nerve fibers of distal colonic mucosa in ulcerative colitis mice].

OBJECTIVE: To observe the effect of moxibustion at "Zusanli" (ST 36) on distal, middle and proximal colonic mucosal injury and expression of calcitonin gene-related peptide (CGRP) positive nerve fibers of distal colonic mucosa in ulcerative colitis (UC) mice at different time points. METHODS: A total of 51 C57BL/6N mice were randomized into a 7-day control group (n=8), a 7-day model group (n=7), a 7-day moxibustion group (n=7), a 14-day control group (n=6), a 14-day model group (n=14) and a 14-day moxibustion group (n=9). In the model groups and the moxibustion groups, 2% dextran sulfate sodium (DSS) was given for 7-day free drinking to establish the UC model. Three days into modeling, moxibustion was applied at "Zusanli" (ST 36) in the 7-day moxibustion group and the 14-day moxibustion group, once a day, 10 min a time for 5 days and 12 days respectively. HE staining was used to observe the morphology of colonic tissue, the percentages of distal, middle and proximal colonic mucosal injury were calculated. Immunofluorescence staining was used to detected the expressions of positive nerve fibers of distal, middle and proximal colonic mucosa and CGRP positive nerve fibers of distal colonic mucosa. RESULTS: Mucosal injury can be observed in mice after modeling, displaying epithelial layer disappearance, abnormal crypt structure or crypt disappearance. Compared with the 7-day control group, colon length was shortened (P<0.001), percentages of overall, distal, middle colonic mucosal injury were increased (P<0.001), the expressions of positive nerve fibers of distal, middle and proximal colonic mucosa and CGRP positive nerve fibers of distal colonic mucosa were increased (P<0.001, P<0.05, P<0.01) in the 7-day model group. Compared with the 7-day model group, the expressions of positive nerve fibers of middle and distal colonic mucosa and CGRP positive nerve fibers of distal colonic mucosa were decreased in the 7-day moxibustion group (P<0.05). Compared with the 14-day control group, the colon length was shortened (P<0.01), percentage of overall colonic mucosal injury was increased (P<0.001) in the 14-day model group. Compared with the 14-day model group, colon length was lengthened (P<0.05), percentage of overall colonic mucosal injury was decreased (P<0.05) in the 14-day moxibustion group. CONCLUSION: Moxibustion at "Zusanli" (ST 36) can reduce the expressions of positive nerve fibers of colonic mucosa and CGRP positive nerve fibers of distal colonic mucosa, thus, improve the colonic mucosal injury.

Synthesis and biological evaluation of oxoindolin-3-ylidene ethyl benzothiohydrazides as non-peptide TPO mimics.

A novel series of oxoindolin-3-ylidene ethyl benzohydrazides were designed, synthesized, and identified as small molecule agonists of thrombopoietin (TPO) receptor c-mpl. Sulfur-oxygen exchange in oxoindolin-3-ylidene ethyl benzohydrazides was found to improve their agonistic activities. Several oxoindolin-3-ylidene ethyl benzothiohydrazides have been identified as full agonists of c-mpl.

[Study on the change of angiotensin II expression in rat lung exposure to acrolein].

OBJECTIVE: To investigate the changes of angiotensin II (Ang II) expression in a rat model of mucus hypersecretion induced by acrolein. METHODS: A rat model of mucous hypersecretion of airway was established by acrolein-inhalation. The AB/PAS stain was performed to evaluate the mucous secretion level. Immunohistochemistry and Western blot were employed to detect the expression of Ang II protein. RESULTS: Hypersecretion of mucus occurred at 3 weeks and peaked at 6 weeks post exposure (wpe) to acrolein. Increased immunostaining and protein level of Ang II were readily detected in rat lung at 3 wpe and enhanced at 6 wpe. CONCLUSION: The increase of Ang II expression, which associated to pulmonary mucus hypersecretion, might play a role in the lung injury induced by acrolein exposure.

[Effects of acrolein exposure on the expression of Muc5ac in the airway of rats].

OBJECTIVE: To explore the changes of Muc5ac expression in a rat model of mucus hypersecretion induced by acrolein. METHODS: The rat model of mucus hypersceretion was established by acrolein exposure. RT-PCR, ISH, immunohistochemistry and Western blot were employed to detect the mRNA and protein levels of Muc5ac in the rats airway. RESULTS: The expression of Muc5ac was detected in trachea and lung at 3 weeks post exposure (wpe) and the expression level was enhanced at 6 wpe. ISH confirmed that Muc5ac was synthesized endogenously. CONCLUSION: The pulmonary mucus hypersecretion is associated to the increase of intrinsic Muc5ac expression in the rat airway exposure to acrolein.

[Effects of enalapril on NF-kappaB expression in mucus hypersecretion rats induced by acrolein exposure].

OBJECTIVE: To investigate the effects of enalapril on nuclear factor-kappa B (NF-kappaB) expression in a rat model of mucus hypersecretion induced by acrolein. METHODS: The rat model of mucus hypersecretion was established by the inhalation of acrolein. Then the mucus hypersecretion rats were treated by enalapril. The AB/PAS stain was performed to measure the secretion of airway in the rats, while the mRNA and protein expression levels of NF-kappaB in the airway were detected by RT-PCR, ISH, immunohistochemistry and Western blot. RESULTS: With the inhalation of acrolein, the airway secretion increased obviously, while the mRNA and protein expressions of NF-kappaB in lung were increased. The treatment of enalapril resulted in a significant decrease of airway secretion, as well as the downregulation of NF-kappaB in both protein and gene level. CONCLUSION: Enalapril (ACE inhibitor) treatment could effectively decrease pulmonary mucus hypersecretion in acrolein-treated rats, which may be related to the downregulation of NF-kappaB expression.

[Effects of enalapril on IL-1beta, IL-6 expression in rat lung exposure to acrolein].

OBJECTIVE: To investigate the effects of enalapril on the expressions of IL-1beta and IL-6 in the lung of rats treated with acrolein inhalation. METHODS: Inflammatory lung injury was induced by acrolein inhalation in rats. The rats were divided into natrium solution (NS) group, acrolein group, enalapril and acroclein (EA) group, enalapril and NS (EN) group. Lungs were harvested from the rats in each group at 1 week, 3 weeks, 6 weeks after the treatment of natrium solution, acrolein inhalation and/or enalapril. The mRNA and protein expressions of IL-1beta and IL-6 in the lung tissues were measured by RT-PCR, in situ hybridization, immunohistochemistry and Western blot. RESULTS: Increased immunostaining, protein level and mRNA expression of IL-1beta and IL-6 were found in rat lung at 3 weeks and reached to the peak at 6 weeks post exposure to acrolein. The administration of enalapril resulted in a significant downregulation of IL-1beta and IL-6 in both protein and gene level, accompany with the decrease of inflammation. CONCLUSION: Enalapril, as a ACE inhibitor, could protect the airway from inflammation injury in acrolein-treated rats via the down-regulation of IL-1beta and IL-6 expression.

[Changes of proteomics in the injured lung of adult rats subjected to acrolein inhalation].

OBJECTIVE: To investigate the proteomics change in injured lung tissues of adult rats subjected to acrolein inhalation for 6 weeks. METHODS: Two-dimensional electrophoresis was used to define the change of proteomic expresses and mass spectrometry was applied to identify the amino acid sequence in differential expressional proteins in lung tissues between acrolein inhalation group and saline control group. RESULTS: A total of 545 protein spots were found in the lung tissues tested. Of the 545 proteins, 7 differentially expressed proteins were detected in acrolein inhalation group as compared with saline control group. Among them, tyrosine 3/tryptophan 5-monooxygenase activation protein, phosphatidylinositol transfer protein, poly(A)-binding protein, presented a marked upregulation, while 2601 and 2603 vimentin, phosphatidylinositol transfer protein, and chloride intracellular channel 3 showed a marked downregulation. CONCLUSION: This study provides the crucial evidence that differential expressional proteins may underline the machenism of lung injury after acrolein inhalation in adult rats.

[Changes of proteomics in the lung of rats subjected to acrolein inhalation after enalapril administration].

OBJECTIVE: To study the molecular mechanism of enalapril on rat chronic bronchopneumonia model with proteomics method. METHODS: Enalapril treatment applied to the recovery of chronic bronchopneumonia in rats, after enalapril administration, two-dimensional electrophoresis and mass spectrometry were performed to analyze the difference of the peptide finger print map and amino acid sequence of rat lung tissues in acrolein inhalation group and enalapril treatment group. RESULTS: Three differential protein spots from average 545 protein spots were found in lung tissues of two groups. These protein spots were identified as glyceraldehyde-3-phosphate dehydrogenase, T-kininogen 1 precursor and dihydropyrimidinase-like 2 with mass spectrometry. The significant upregulation in the level of dihydropyrimidinase-like 2, downregulation of glyceraldehyde-3-phosphate dehydrogenase, and the lose of T-kininogen 1 precursor in enalapril treatment group were observed. CONCLUSION: The expression difference in three proteins might contribute to the preventive role of enalapril on acrolein inhalation induced rat chronic bronchopneumonia.

Diagnostic value of computerized nuclear morphometry for the prediction of malignancy in liver fine needle aspiration cytology.

OBJECTIVE: To evaluate the diagnostic value of nuclear parameters using computerized morphometry in the differential diagnosis of well-differentiated hepatocellular carcinoma (WD-HCC) and regenerative atypia in liver fine needle aspiration cytology (FNAC). STUDY DESIGN: All 78 cases, including 30 negative, 30 positive and 18 suspicious liver FNA cytology smears were examined with regard to nuclear area, nuclear perimeter, largest to smallest diameter ratio of the nuclei (L/S ratio) and coefficient of variation of the nuclear area (NACV). The diagnostic value was assessed using receiver operating characteristics (ROC) curves. Cutoff points were determined by the area under the ROC curve (AUC), which provides the highest combined sensitivity and specificity. RESULTS: The nuclear area and perimeter in liver FNA were more helpful than L/S ratio and NACV in discriminating between hepatocytes with regenerative atypia and WD-HCC. Cutoff points of 56.67 for nuclear area and 27.16 for perimeter were suggested to help shift the suspicious diagnoses to WD-HCC. CONCLUSION: When faced with difficult diagnoses on liver FNA, we suggest application of computerized morphometry may be a helpful ancillary diagnostic tool in the further classification of such cases.

[Improvement and application of diagnostic techniques of the skeletal muscle biopsy].

OBJECTIVE: To summarize the improvement of various muscle staining techniques and discuss their application in diagnosis of neuromuscular diseases. METHODS: Three hundred cases of skeletal muscle biopsy samples were examined by histopathological methods. The flash-freezing techniques were used for the preparation of frozen section, which were stained with HE, Gomori trichromic (GMR), glycogen (PAS), and fat acid (oil red O); the enzyme histochemical staining with myosin adenosine triphosphatase (ATPase), and NADH-TR. Those stained methods had been improved. The immunohistochemical staining with dystrophin; and transmission electronic microscopy were used. RESULTS: Deepfreeze with heteropentane-liquid nitrogen and flash-freezing techniques could avoid artifacts of ice crystal vacuolation. GMR staining mainly showed the degenerative and necrotic lesions of mitochondria and muscle fibers. Oil red O staining showed the increase of lipid in muscle fibers. PAS staining mainly showed glycogen and glycoprotein. Application of frozen sections in muscular tissue was better than that of paraffin sections. NADH-TR staining and ATPase staining could distinguish the two types of muscle fibers, and show the changes of inner structure of muscle fibers and mitochondria enzymes. CONCLUSION: The distribution and characteristic pathological changes of the two types of muscle fibers can be showed clearly by enzyme and non-enzyme histochemical staining techniques of the skeletal muscle. These methods can compliment with each other. Only after understanding the technical principles can we master and apply these methods.