Phylogenetic Conservation of Soil Microbial Responses to Elevated Tropospheric Ozone and Nitrogen Fertilization.

Plant primary productivity and crop yields have been reduced due to the doubled level of global tropospheric ozone. Little is known about how elevated ozone affects soil microbial communities in the cropland ecosystem and whether such effects are sensitive to the nitrogen (N) supply. Here, we examined the responses of bacterial and fungal communities in maize soils to elevated ozone (+60 ppb ozone) across different levels of N fertilization (+60, +120, and +240 kg N ha-1yr-1). The fungal alpha diversity was decreased (P < 0.05), whereas the bacterial alpha diversity displayed no significant change under elevated ozone. Significant (P < 0.05) effects of N fertilization and elevated ozone on both the bacterial and fungal communities were observed. However, no interactive effects between N fertilization and elevated ozone were observed for bacterial and fungal communities (P > 0.1). The bacterial responses to N fertilization as well as the bacterial and fungal responses to elevated ozone were all phylogenetically conserved, showing universal homogeneous selection (homogeneous environmental conditions leading to more similar community structures). In detail, bacterial Alphaproteobacteria, Actinobacteria, and Chloroflexi, as well as fungal Ascomycota, were increased by elevated ozone, whereas bacterial Gammaproteobacteria, Bacteroidetes, and Elusimicrobia, as well as fungal Glomeromycota, were decreased by elevated ozone (P < 0.05). These ozone-responsive phyla were generally correlated (P < 0.05) with plant biomass, plant carbon (C) uptake, and soil dissolved organic C, demonstrating that elevated ozone affects plant-microbe interactions. Our study highlighted that microbial responses to elevated ozone display a phylogenetic clustering pattern, suggesting that response strategies to elevated ozone stress may be phylogenetically conserved ecological traits. IMPORTANCE The interactions of plant and soil microbial communities support plant growth and health. The increasing tropospheric ozone decreases crop biomass and also alters soil microbial communities, but the ways in which crops and their associated soil microbial communities respond to elevated tropospheric ozone are not clear, and it is also obscure whether the interactions between ozone and the commonly applied N fertilization exist. We showed that the microbial responses to both elevated ozone and N fertilization were phylogenetically conserved. However, the microbial communities that responded to N fertilization and elevated ozone were different, and this was further verified by the lack of an interactive effect between N fertilization and elevated ozone. Given that the global tropospheric ozone concentration will continue to increase in the coming decades, the decrease of specific microbial populations caused by elevated ozone would result in the extinction of certain microbial taxa. This ozone-induced effect will further harm crop production, and awareness is urgently needed.

[Effects of Greenhouse and Open-field Cultivation on Heavy Metal Uptake During Carrot Growth].

Clarifying the absorption dynamics of heavy metal(loid)s by crops under different cultivation methods is critical for risk management and control for heavy metal pollution. Here, taking carrots as an example, the pH, cation exchange capacity (CEC), and contents of heavy metals in soil and carrots were analyzed. We compared the absorption and transport characteristics of six metal(loid)s (As, Cd, Cr, Cu, Pb, and Zn) during the three key growth periods under greenhouse and open-field cultivation methods. In addition, the effects of planting methods on carrot biomass and heavy metal content over time were studied, and a health risk assessment was conducted. The results showed that the greenhouse and open-field cultivation methods had the following in common:① As the carrots continuously grew, the metal uptake and biomass in the belowground part (edible part) and the metal(loid) concentrations in the aboveground and belowground parts both showed trends of increasing first and then decreasing or stabilizing. ② The absorption of As, Cd, Cr, and Pb in carrots was mostly accumulated in the aboveground part. ③ The content of Cd in the edible part exceeded the standard, and the total target hazard quotient was>1, indicating potential adverse health risks, most of which were contributed by As and Cd. Compared to that under open-field, the short-term greenhouse cultivation had a harvest time approximately 15 d earlier. The As, Cd, and Pb concentrations in the aboveground part during the seedling and fast-growing periods were significantly lower in the greenhouse than that in the open-field (P<0.05) but did not differ between greenhouse and open-field (except As) at the mature stage. The concentrations of As and Cd in the belowground part of greenhouse carrots were lower in the greenhouse than those of carrots grown in the open-field at the seedling stage (P<0.05). The absorption of As and Cd was decreased significantly from 0 to 95 d (P<0.05), and there was no significant difference in the metal concentration and absorption in the rest of the growth stages. The bioconcentration factors (BCF) of As, Cd, and Pb in the aboveground part in all stages and those of As and Cd in the belowground part in the seedling stage were lower in the greenhouse than that in the open-field. This study emphasizes the possibility of short-term changes in cultivation mode to reduce the risks of metal(loid)s in contaminated farmland vegetables.

Molecular characterization of methanogenic microbial communities for degrading various types of polycyclic aromatic hydrocarbon.

Knowledge on methanogenic microbial communities associated with the degradation of polycyclic aromatic hydrocarbons (PAHs) is crucial to developing strategies for PAHs bioremediation. In this study, the linkage between the type of PAHs and microbial community structure was fully investigated through 16S rRNA gene sequencing on four PAH-degrading cultures. Putative degradation products were also detected. Our results indicated that naphthalene (Nap)/2-methylnaphthalene (2-Nap), phenanthrene (Phe) and anthracene (Ant) sculpted different microbial communities. Among them, Nap and 2-Nap selected for similar degrading bacteria (i.e., Alicycliphilus and Thauera) and methanogens (Methanomethylovorans and Methanobacterium). Nap and 2-Nap were probably activated via carboxylation, producing 2-naphthoic acid. In contrast, Phe and Ant shaped different bacterial and archaeal communities, with Arcobacter and Acinetobacter being Phe-degraders and Thiobacillus Ant-degrader. Methanogenic archaea Methanobacterium and Methanomethylovorans predominated Phe-degrading and Ant-degrading culture, respectively. These findings can improve our understanding of natural PAHs attenuation and provide some guidance for PAHs bioremediation in methanogenic environment.

Effects of biochar on bacterial communities in a newly established landfill cover topsoil.

Recent studies revealed the benefits of applying biochar in landfill final cover soil, such as adsorbing odorous compounds and promoting microbial methane oxidation. Most of these processes are related to the soil bacterial communities. However, the effects of biochar application on the overall bacterial community in newly established landfill cover soil are not yet understood, especially in field condition. The objective of the present field study is to investigate the effects of biochar on the diversity of soil bacterial community 3 months after incubation (short-term). Landfill final cover topsoil (0.6 m) was amended with 0 (control), 5, and 10% (w/w) of biochar derived from peanut-shell and wheat straw. Soil bacterial communities were analysed using the 16S rRNA-based T-RFLP approach. Biochar application significantly (p < 0.05) increased the diversity of soil bacterial communities. The Shannon diversity index of bacterial communities in soil amended with 5 and 10% of biochar was increased from 3.34 to 3.85 and 3.92, respectively. There were four bacterial phyla recorded found at both control and amended soils, namely Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria. In addition, Gemmatimonadetes was found only in soil amended with 10% biochar. The interactions between soil bacterial communities and measured soil parameters including moisture content, electrical conductivity, total organic matter, total nitrogen and total phosphorus were found to be statistically non-significant (p > 0.05), according to the canonical correspondence analysis (CCA). This may be due to the highly heterogeneous nature of landfill soil. Results from this study revealed that short-term biochar application already altered the soil physicochemical properties and increased the diversity of soil bacterial communities.

PBDEs and Dechlorane Plus in the environment of Guiyu, Southeast China: A historical location for E-waste recycling (2004, 2014).

Polybrominated diphenyl ethers (PBDEs, tri-to hepta-BDEs) and Dechlorane Plus (DP) in multiple samples (soil, sediment, road dust, and PM2.5 particles) in historical locations of our previous work in Guiyu (electronic-waste recycling town) in southeastern China were investigated in 2014. Ten years later, PBDEs and DP were detected in 100% of the samples. Æ©3-7PBDEs were still relatively great, ranging from 1.2 × 101 to 2.1 × 103 ng/g dry weight (dw) in soil, 2.1 to 3.2 × 103 ng/g dw in sediment, 1.0 × 101 to 1.1 × 104 ng/g dw in road dust, which were a little less than or close to those measured in 2004. However, Æ©3-7PBDEs in PM2.5 (5.0 × 102 to 8.4 × 102 pg/m3) were significantly lower in 2014. BDE-47, -99, -153, and -183 were predominant congeners, which were also predominant PBDEs reported in Guiyu (2006-2008). Greater proportions of lesser-brominated BDEs were predominant in PM2.5 than other samples. DP was detected in 100% of the samples collected with high levels. Total syn-DP and anti-DP concentrations were 3.8 to 2.1 × 103 ng/g dw in soil, 1.1 × 103 to 7.2 × 103 ng/g dw in sediment, 1.4 × 101 to 1.1 × 103 ng/g dw in road dust, and 1.8 × 102 and 1.7 × 102 pg/m3 in PM2.5. Most of the fractions of anti-DP (fanti) (70%-80%) were consistent among samples. The health risk assessment showed that hazard indexes (HIs) of PBDEs or DPs for child or adult were all lower than 0.16.

Comparison of plant and bacterial communities between a subtropical landfill topsoil 15years after restoration and a natural area.

Engineered sanitary landfills are becoming more and more common worldwide. Ecosystem restoration of capped sanitary landfills is essential to restore the disturbed environment. Comparing plant communities, as well as bacterial communities, in landfills and natural areas, offers an efficient way to assess the restoration status. However, such studies on the restored engineered landfills are limited. Here we present an ecological restoration case in an engineered landfill in a subtropical region. Part of the South East New Territories (SENT) landfill in Hong Kong was capped and restored, by using 16 plant species growing on top of the final cover soil, during 1997-1999. In 2014, plant survey and soil properties analyses were conducted in a restored site (AT) and a natural site (CT, an undisturbed area, serving as a control). The similarity between the biota communities (i.e., plant and soil bacteria) of the two sites was assessed. Plant and soil bacterial communities at AT were significantly different (R=1, P<0.01, ANOSIM) from those at CT. A lower plant diversity but a higher soil bacterial diversity were observed at AT. The soil bacterial community structure was potentially driven by soil pH, moisture content, cation exchange capacity (CEC), N, and P. The engineered landfill had not been restored to an ecosystem similar to the natural environment 15years after restoration. Establishing similar soil properties in the landfill topsoil would be important to achieve a bacterial community similar to the natural area. This study can also offer a quick and inexpensive method for landfill engineers to assess the bacterial restoration of man-made ecosystems using plant and soil properties rather than DNA analyzing techniques.

Mycorrhizal colonization status of lowland rice (Oryza sativa L.) in the southeastern region of China.

The accumulation, distribution, and speciation of contaminants, such as arsenic, in rice can be affected by soil microorganisms such as arbuscular mycorrhizal fungi (AMF). As a potential measure to control contaminant acquisition in rice, the status and performance of AMF in the field need to be investigated. Root samples of rice plants were collected in seven different cities in Guangdong, Jiangxi, Hubei, and Jiangsu Provinces in China in order to investigate the colonization rate of AMF. The total DNA of the roots was extracted, followed by PCR and sequencing, and further confirmed the existence of AMF. The highest colonization rates (19.5 ± 7.2%) were observed in samples from Huizhou City, Guangdong Province. Sequences of ribosomal DNA derived from Pingtan (PT) and Shuikou (SK) in Huizhou shared a similarity of 73 and 86% to Glomus cf. clarum Att894-7 (FM865542) and "uncultured fungus" (EF434122.1), respectively. The moisture tolerance of the AMF from different sources was tested by subjecting to different levels of water content in the soil. Only AMF from PT, SK, and LJ colonized rice under a condition of 100% of the soil water holding capacity (WHC), but not those isolated from upland plants. The AM colonization rate could be governed by the lighting conditions and temperature. AMF isolated in paddy fields has been shown to have more tolerance to moisture than other upland species. Radial oxygen loss (species and stress dependent) could be an essential factor influencing the colonization rate and requires more investigation.

Measurement and health risk assessment of PM2.5, flame retardants, carbonyls and black carbon in indoor and outdoor air in kindergartens in Hong Kong.

Indoor air pollution is closely related to children's health. Polybrominated diphenyl ethers (PBDEs) and dechlorane plus (DP) transmitted through indoor PM2.5 and dust, along with carbonyl compounds and black carbon (BC) aerosol were analysed in five Hong Kong kindergartens. The results showed that 60% of the median PM2.5 levels (1.3×101 to 2.9×101µg/m3 for indoor; 9.5 to 8.8×101µg/m3 for outdoor) in the five kindergartens were higher than the guidelines set by the World Health Organization (2.5×101µg/m3). Indoor PM2.5 mass concentrations were correlated with outdoor PM2.5 in four of the kindergartens. The PBDEs (0.10-0.64ng/m3 in PM2.5; 0.30-2.0×102ng/g in dust) and DP (0.05-0.10ng/m3 in PM2.5; 1.3-8.7ng/g in dust) were detected in 100% of the PM2.5 and dust samples. Fire retardant levels in the air were not correlated with the levels of dust in this study. The median BC concentrations varied by >7-fold from 8.8×102ng/m-3 to 6.7×103ng/m-3 and cooking events might have caused BC concentrations to rise both indoors and outdoors. The total concentrations of 16 carbonyls ranged from 4.7×101µg/m3 to 9.3×101µg/m3 indoors and from 1.9×101µg/m3 to 4.3×101µg/m3 outdoors, whilst formaldehyde was the most abundant air carbonyl. Indoor carbonyl concentrations were correlated with outdoor carbonyls in three kindergartens. The health risk assessment showed that hazard indexes (HIs) HIs of non-cancer risks from PBDEs and DPs were all lower than 0.08, whilst non-cancer HIs of carbonyl compounds ranged from 0.77 to 1.85 indoors and from 0.50 to 0.97 outdoors. The human intake of PBDEs and DP through inhalation of PM2.5 accounted for 78% to 92% of the total intake. The cancer hazard quotients (HQs) of formaldehyde ranged from 4.5E-05 to 2.1E-04 indoors and from 1.9E-05 to 6.2E-05 outdoors. In general, the indoor air pollution in the five Hong Kong kindergartens might present adverse effects to children, although different schools showed distinct pollution levels, so indoor air quality might be improved through artificial measures. The data will be useful to developing a feasible management protocol for indoor environments.

Feasibility of biochar application on a landfill final cover-a review on balancing ecology and shallow slope stability.

Due to the increasing concerns on global warming, scarce land for agriculture, and contamination impacts on human health, biochar application is being considered as one of the possible measures for carbon sequestration, promoting higher crop yield and contamination remediation. Significant amount of researches focusing on these three aspects have been conducted during recent years. Biochar as a soil amendment is effective in promoting plant performance and sustainability, by enhancing nutrient bioavailability, contaminants immobilization, and microbial activities. The features of biochar in changing soil physical and biochemical properties are essential in affecting the sustainability of an ecosystem. Most studies showed positive results and considered biochar application as an effective and promising measure for above-mentioned interests. Bio-engineered man-made filled slope and landfill slope increasingly draw the attention of geologists and geotechnical engineers. With increasing number of filled slopes, sustainability, low maintenance, and stability are the major concerns. Biochar as a soil amendment changes the key factors and parameters in ecology (plant development, soil microbial community, nutrient/contaminant cycling, etc.) and slope engineering (soil weight, internal friction angle and cohesion, etc.). This paper reviews the studies on the production, physical and biochemical properties of biochar and suggests the potential areas requiring study in balancing ecology and man-made filled slope and landfill cover engineering. Biochar-amended soil should be considered as a new type of soil in terms of soil mechanics. Biochar performance depends on soil and biochar type which imposes challenges to generalize the research outcomes. Aging process and ecotoxicity studies of biochar are strongly required.

Risk assessment of arsenic and other metals via atmospheric particles, and effects of atmospheric exposure and other demographic factors on their accumulations in human scalp hair in urban area of Guangzhou, China.

Eighty-eight scalp hair samples were collected from Guangzhou (GZ) urban population (15-65 years) to investigate the accumulation of As and other metals (Cr, Mn, Ni, Cu, Zn, Cd, Sn, Sb, Hg and Pb). Demographic information, including body weight, height, age, gender, habits of smoking and drinking, types of drinking water, duration of stay in GZ, days of stay in GZ per year (days/year), and hours spent in indoor environment per day (h/day), were also recorded during hair sampling to refine the uncertainty of risk assessment derived from exposures to elements via dust and airborne particles. No significant non-carcinogenic risk was found. However, the cancer risks of Cr and As for both ingestion and inhalation exceeded the most tolerable regulated level (1.0×10(-6)). The environmental exposures to urban dust and airborne particles were observed significantly correlated to accumulations of Cd (R=0.306, p=0.005) and Ni (R=0.333, p=0.002) in scalp hair. Furthermore, the hair burden of elements was also significantly (p<0.05) dependent on gender (Mn, Ni, Zn, As, Sn and Hg), age (Cr, As, Cd and Hg), duration of stay in GZ (Hg) as well as nutritional and physical status, reflected by BMI and BSA (Cr, Ni, Cd, Sb and Hg). Nutritional and physical status was observed as the exclusive important factor influencing As speciation in human scalp hair. However, habits of smoking and alcohol drinking as well as types of drinking water were not identified as the significant influencing factors on any element (p>0.05).

Arsenic speciation in total contents and bioaccessible fractions in atmospheric particles related to human intakes.

Speciation of inorganic trivalent arsenicals (iAs(III)), inorganic pentavalent arsenicals (iAs(V)), monomethylarsonic acid (MMA) and dimethylarsinic acid (DMA) in total arsenic (As) content and its bioaccessible fractions contained in road dust, household air-conditioning (AC) filter dust and PM2.5 was investigated. Inorganic As, especially iAs(V), was observed as the dominant species. Physiologically based extraction test (PBET), an in-vitro gastrointestinal method, was used to estimate the oral As bioaccessibility in coarse particles and the species present in the oral bioaccessible fraction. A composite lung simulating serum was used to mimic the pulmonary condition to extract the respiratory bioaccessible As and its species in PM2.5. Reduction of iAs(V) to iAs(III) occurred in both in-vitro gastrointestinal and lung simulating extraction models. The inorganic As species was the exclusive species for absorption through ingestion and inhalation of atmospheric particles, which was an important exposure route to inorganic As, in addition to drinking water and food consumption.

Arsenite transporters expression in rice (Oryza sativa L.) associated with arbuscular mycorrhizal fungi (AMF) colonization under different levels of arsenite stress.

As a silicon hyperaccumulator, lowland rice takes up higher levels of As than many other plants due to silicic acid and arsenite sharing the same transporters (Lsi1 and Lsi2). Glomus intraradices (AH01) was inoculated to rice under different arsenite concentrations (0, 2 and 8 µM) in order to investigate the interactions between arbuscular mycorrhizal fungus and rice on the accumulation of arsenite. The relative mRNA expressions of Lsi1 and Lsi2 resulted in a down-regulating trend in mycorrhizal plants. Under 2 µM arsenite treatments, Lsi1 and Lsi2 were significantly decreased, by 0.7-fold (P<0.05) and 0.5-fold (P<0.01), respectively, in mycorrhizal plants when compared with non-mycorrhizal plants. This led to the decrease of arsenite uptake per unit of root dry mass. No organic As species were detected in both roots and shoots. The As(III)/As(V) ratios indicated that mycorrhizal plants immobilized most of the arsenite proportion in the roots and prevented its translocation from the roots to the shoots.

[Effect of reinforcing Qi strength spleen in the expression of bFGF and EGF in treating serious soft tissue injury].

OBJECTIVE: Using the method of bleeding from the orbital vein and lancing to make the animal model of trauma, and to observe the influence of reinforcing Qi strength spleen in the expression of bFGF and EGF in the reparative process of raw surface, in order to explore the possible mechanism of reinforcing Qi strength spleen in promoting the rehabilitation of soft tissue. METHODS: Forty healthly adult SD rats were made to be traumatic model using the method of bleeding from the orbital vein and lancing. After operation, there were 33 rats survival, which were divided into the reinforcing Qi strength spleen group, the activating blood circulation to dissipate blood stasis group and the model group randomly. The raw surface and ambient normal skin were taken at the 3rd, 7th and 14th days after operation to detect the expression of bFGF and EGF by immunohistochemical method. RESULTS: At the 3rd, 7th and 14th days after operation, the expression of bFGF and EGF in the tissue of raw surface of the reinforcing Qi strength spleen group and the activating blood circulation to dissipate blood stasis group was obviously higher than that of the model group(P < 0.05). Compared with the activating blood circulation to dissipate blood stasis group, the expression of bFGF and EGF in the tissue of raw surface of the reinforcing Qi strength spleen group was higher (P < 0.05) in the 3rd and 7th day after operation. But in the 14th after operation, there was no significantly difference between reinforcing Qi strength spleen group and activating blood circulation to dissipate blood stasis group. CONCLUSION: The method of reinforcing Qi strength spleen can efficiently promote the expression of bFGF and EGF in raw surface of serious soft tissue injury.

[Clinical study of transforaminal instrumented lumbar interbody fusion].

OBJECTIVE: To study the effect and complications of transforaminal lumbar interbody fusion technique. METHODS: The medical records and radiographs of 40 patients undergone transforaminal lumbar interbody fusion between 2005 and 2007 were retrospectively reviewed. There were 49 segments with fusion. Preoperative and 1 year postoperative functional evaluation were graded with ODI and VAS scoring system. The height and angle of the intervertebral space and the fusion status were measured as well. RESULTS: All patients were followed-up for 12 to 24 months with the average of 18 mouths. There were no severity postoperative complications. The operation time averaged 160 min and average blood loss 510 ml. The effect results were excellent in 28 cases, good in 7 and fair in 5. The fusion rate was 100%. One year after operation, the pain relief in the VAS and the reduction of the ODI were significant (P < 0.01), the height and angle of the intervertebral space increased obviously (P < 0.05). Fifteen patients complained low back pain to some extent untill the last follow-up. CONCLUSION: Transforaminal lumbar interbody fusion can achieve satisfactory clinical and radiographic results especially for the failed back surgery syndrome.

[Curative effect analysis of skin flap and allogeneic tendon in reconstructing limbs function of complex soft-tissue defect].

OBJECTIVES: To investigate the curative effect of skin flap and allogenic tendon in reconstructing limbs function of complex soft-tissue defect. METHODS: From May 2001 to December 2007, 43 cases of complex soft-tissue defect of limbs were repaired by pedicled skin flaps, free skin flaps, cutaneous nerve nutrient vessel skin flaps and arterial island skin flaps for primary stage, then the function of the limbs were reconstructed with allogeneic tendon after 2-3 months of skin flap operation. There were 31 males and 12 females, aged 5-53 years (mean 25 years). Injury was caused by machine in 28 cases, by traffic accident in 14 cases and others in 1 case. There were 27 cases in upper limb, 16 cases in lower limb. Twenty-six cases complicated by bone fracture, dislocation and bone defect, the most of bone defect were cortical bone defect. The sizes of skin and parenchyma defect were 9 cm x 4 cm to 37 cm x 11 cm, the length of tendon defect was 6 to 26 cm. The sizes of skin flaps were 10 cm x 5 cm to 39 cm x 12 cm. Allogeneic tendons were used 2-6 strips (mean 4 strips). RESULTS: Forty-three cases were followed up for 5-56 months (16 months on average), all flaps survived. The donor area healed by first intention, the incision healed by first intention in second operation, and no tendon rejection occurred. The clinical healing time of fracture was 3-8 months, and the clinical healing time of allograft was 6-8 months. Six cases were given tenolysis for adhesion of tendon after 3-5 months of tendon transplantation. The postoperative flexion of wrist joint was 20-50 degrees, the extension was 20-45 degrees. Articulation metacarpophalangeal and articulation interphalangeal could extend completely. The flexion of articulation metacarpophalangeal of thumb was 20-45 degrees, the flexion of articulation interphalangeal was 30-70 degrees. The flexion of articulation metacarpophalangeal and articulation interphalangeal of the other fingers was 60-90 degrees. The postoperative ankle can extend to neutral position, the neutral position of ankle was 30-50 degrees. The flexion of articulation metatarsophalangeal and articulation interdigital was 20-40 degrees. The extension of articulation metatarsophalangeal was 30-60 degrees. CONCLUSION: Through designing systematically treatment plan, practicing operation by stages, preventing adhesion of tendon actively and exercising function reasonably, the functions of limbs reconstructed by allogenic tendon and skin flap can leads to satisfactory effect.

[Experimental study on the treatment of serious soft tissue injuries with strengthening the spleen and replenishing qi].

OBJECTIVE: To study the effects of Chinese drugs based on strengthening the spleen and replenishing qi treatment rule on neoformative capillaries and fibroblast during the soft tissue repair after serious trauma in rats, so as to explore the biological basis of the TCM theory "the spleen dominate extremities and muscles" applied to the treatment of soft tissue injuries. METHODS: The model rats were established by bleeding from femoral artery and lancing method, and the rats were randomly divided into the control group, strengthening the spleen group and activating blood and resolving stasis group. The samples were got from the tissue of the wounded area at the 5th, 10th and 15th days after oral administration of the traditional Chinese medicine. After fixation and section, the tissues were stained by CD31 and PCNA staining. The amount of the capillaries and fibroblasts in the tissue of the wounded area were observed through multi-purpose microscope (ZEISS Axioskop2). Quantitative analysis was carried out on Image-ProPlus image analyzer. RESULTS: The amount of the capillaries and fibroblasts in the wounded tissue in the strengthening the spleen group were larger than that in the control group at the 5th, 10th and 15th day. And the proliferation speed of capillaries and fibroblasts was faster than those in the control group or the activating blood and resolving stasis group. CONCLUSION: The Chinese drugs according to strengthening the spleen and replenishing qi treatment rule were effective to promote growth of the granulation tissue and facilitate healing of the wounded area. And it has better effect than the treatment of promoting blood circulation and removing stasis.

Osmic acid staining of myelin sheath in normal and regenerated peripheral nerves.

OBJECTIVE: To introduce a practical, economical, and time-saving method to stain (with osmic acid) the myelin sheath in normal and regenerated peripheral nerves. METHODS: A total of 12 Sprague Dawley rats, weighing 250-320 g (mean equal to 276 g+/-38 g), were divided into two groups: a normal nerve group (n equal to 6) and a regenerated nerve group (n equal to 6). In the normal nerve group, the ventral and dorsal roots of L(4) to L(6) and their sciatic nerves were harvested for histological analysis. While in the regenerated nerve group, the right sciatic nerves were severed and then repaired with an epineurial microsuture method. The repaired nerves were harvested 12 weeks postoperatively. All the specimens were fixed in 4% paraformaldehyde and transferred to 2% osmic acid for 3-5 days. Then the specimens were kept in 75% alcohol before being embedded in paraffin. The tissues were cut into sections of 3 micromolar in thickness with a conventional microtome. RESULTS: Under a light microscope, myelin sheaths were clearly visible at all magnifications in both groups. They were stained in clear dark colour with a light yellow or colorless background, which provided high contrast images to allow reliable morphometric measurements. Morphological assessment was made in both normal and regenerated sciatic nerves. The ratios of the myelin area to the fibre area were 60.28%+/-7.66% in the normal nerve group and 51.67%+/-6.85% in the regenerated nerve group, respectively (P less than 0.01). CONCLUSIONS: Osmic acid staining is easy to perform and a very clear image for morphometrical assessment is easy to obtain. Therefore, it is a reliable technique for quantitative evaluation of nerve morphology.

[Repair and function reconstruction of complex soft tissue defect of posterior of hand and forearm].

OBJECTIVE: To study the repair and function reconstruction of complex soft tissue defect of posterior of hand and forearm. METHODS: From May 2001 to November 2003, 8 cases of soft tissue defect of posterior of hand and forearm were repaired with thoracico abdominal flaps with hilum for primary stage. The tendon transplantation and allogeneic tendon function reconstruction of hand were performed for secondary stage. The range of the flap was 9 cm x 15 cm to 12 cm x 38 cm. Allogeneic tendon amounted to 6. RESULTS: All the flaps survived. The flap countour was good. The results of allogeneic tendon transplantation were satisfactory and the function of hand was good. CONCLUSION: Repairing complex soft tissue defect of posterior of hand and forearm and reconstructing hand function by use of thoracico abdominal flaps with hilum and transplantation of allogeneic tendon have the satisfactory clinical results.