Promoting nitrogen conversion in aerobic biotransformation of swine slurry with the co-application of manganese sulfate and biochar.

This study aimed to explore the co-application of MnSO4 (Mn) and biochar (BC) in nitrogen conversion during the composting process. A 70-day aerobic composting was conducted using swine slurry, supplemented with different levels of Mn (0, 0.25%, and 0.5%) and 5% BC. The results demonstrated that the treatment with 0.5MnBC had the highest levels of NH4+-N (3.07 g kg-1), TKN (29.90 g kg-1), and NO3--N (1.94 g kg-1) among all treatments. Additionally, the 0.5MnBC treatment demonstrated higher urease, protease, nitrate reductase, and nitrite reductase activities than the other treatments, with the peak values of 18.12, 6.96, 3.57, and 15.14 mg g-1 d-1, respectively. The addition of Mn2+ increased the total organic nitrogen content by 29.59%-47.82%, the acid hydrolyzed ammonia nitrogen (AN) content by 13.84%-57.86% and the amino acid nitrogen (AAN) content by 55.38%-77.83%. The richness of Chloroflexi and Ascomycota was also enhanced by the simultaneous application of BC and Mn. Structural equation modeling analysis showed that Mn2+ can promote the conversion of Hydrolyzed Unknown Nitrogen (HUN) into AAN, and there is a positive association between urease and NH4+-N according to redundancy analysis. Firmicutes, Basidiomycota, and Mortierellomycota showed significant positive correlations with ASN, AN, and NH4+-N, indicating their crucial roles in nitrogen conversion. This study sheds light on promoting nitrogen conversion in swine slurry composting through the co-application of biochar and manganese sulfate.

Insight into the humification and carbon balance of biogas residual biochar amended co-composting of hog slurry and wheat straw.

The impact of biogas residual biochar (BRB) on the humification and carbon balance process of co-composting of hog slurry (HGS) and wheat straw (WTS) was examined. The 50-day humification process was significantly enhanced by the addition of BRB, particular of 5% BRB, as indicated by the relatively higher humic acid content (67.28 g/kg) and humification ratio (2.31) than other treatments. The carbon balance calculation indicated that although BRB addition increased 22.16-46.77% of C lost in form of CO2-C, but the 5% BRB treatment showed relatively higher C fixation and lower C loss than other treatments. In addition, the BRB addition reshaped the bacterial community structure during composting, resulting in increased abundances of Proteobacteria (25.50%) during the thermophilic phase and Bacteroidetes (33.55%) during the maturation phase. Combined these results with biological mechanism analysis, 5% of BRB was likely an optimal addition for promoting compost humification and carbon fixation in practice.

[FTIR microspectroscopic investigation of the age-related changes of subchondral bone of the knee in guinea pig].

Fourier transform infrared (FTIR) microspectroscopy was applied to in-situ analyse the chemical change of tibial articular subchondral bone of female Hartley guinea pigs with age increase. Three infrared absorption regions (a, b, c) of trabecular bone and central marrow region of the subchondral bone were measured for guinea pigs of different ages (1 months, 2 months and 3 months) using the infrared spectrum. Results show that (1) with months increasing, the total area of trabecular bone is increasing, meanwhile, the region a which is similar to normal trabecular bone spectra is decreasing, and region d waveform has the same trend as region a. (2) In the second and third month, region b & c show amide Ill redshift and the red shift in region c shows a shoulder peak, showing the absorption peak intensity on behalf of nucleic acid and polysaccharide in region b & c is 7 times that in region a. (3) beta glycosidic bond absorption peak appears at region c in 3 different old pigs. (4) I(amide III) / I(amide II) is the highest in region b in the second month but lowest in the third month; I(amide III) / I(amide II) reduces from a to c in the second and third month; I(upsilon3)PO2- / I(amide II) in region b & c is 7 times higher than region a in the second and third Month These results are consistent with the regular pattern of change rule of osteoarthritis subchondral bone's organization structure and chemical composition in different stages. Our primary result illustrated that FTIR microspectroscopy can be used for in-situ analysis of the molecular organization of subchondral trabecular bone and bone marrow. It provides reliable pathology information for osteoarthritis subchondral bone tissue at molecular level.

Innovative accumulative risk assessment strategy of co-exposure of As and Pb in medical earthworms based on in vivo-in vitro correlation.

Recent studies on risks assessment of heavy metal(loid) are usually based on their total concentrations. Nevertheless, such an analysis does not assess their real amounts absorbed by human body. To scientifically assess the health risks, in this study medical earthworms were analyzed for relative bioavailability (RBA) of arsenic (As) and lead (Pb) using a multiple gavage mouse model with liver, kidneys, brain, and leg bones as biomarkers for the first time. Metal(loid) bioaccessibility was determined using in vitro physiologically based extraction (PBET) assay. We are the first to develop a novel accumulative health risk assessment strategy by combinational analyzing bioavailability of heavy metal(loid) levels to calculate target organ toxicity dose (TTD) modification of the HI and total cancer risk (TCR), which has capacity to evaluate the health risks of co-exposure of Pb and As in medical earthworms. As a result, As-RBA ranged from 7.2% to 45.1%, and Pb-RBA ranged from 16.1% to 49.8%. Additionally, As and Pb bioaccessibility varied from 6.7% to 48.3% and 7.8% to 52.5%, respectively. Moreover, strong in vivo-in vitro correlations (IVIVCs) were observed between metal-RBA and bioaccessibility, indicating the robustness of the in vitro PBET assay to predict metal-RBA in medical earthworms. The refined accumulative assessment strategy revealed that when adjusted by heavy metal(loid) bioavailability, the TTD modification of HI method typically exhibited an acceptable health risk caused by the co-exposure of Pb and As for cardiovascular, hematological, neurological, and renal system. The TCR levels associated with exposure to Pb and As due to the ingestion of medical earthworms were also acceptable after adjustment by bioavailability. Collectively, our innovation on accumulative risk assessment based on in vivo-in vitro correlation provides a novel approach engaging in assessing the risks due to co-exposure of As and Pb in medical earthworms.

5-Carb-oxy-1,3-bis-(carb-oxy-meth-yl)-4-imidazolinium-4-carboxyl-ate.

The title compound, C(9)H(8)N(2)O(8), was obtained by the reaction of imidazole-4,5-dicarb-oxy-lic acid and 2-chloro-acetic acid. An intra-molecular O-H⋯O hydrogen bond occurs. The crystal packing is stabilized by intermolecular O-H⋯O and C-H⋯O hydrogen bonds, which link mol-ecules into a three-dimensional network.

[Investigation of articular cartilage using FTIR microspectroscopy for guinea pig with spontaneous osteoarthritis].

In-situ analysis the chemical composition of tibial articular cartilage of female Hartley guinea pigs with Fourier transform infrared (FTIR) microspectroscopy was conducted. The infrared spectrum survey consists of three ages (1 months, 2 months and 3 months) and three cartilage layers (surface, middle and deep). The results demonstrated that with ages increasing, the peak positions of main absorbance bands in surface and middle shifted to a lower wavenumber, and in deep they shifted to a lower wavenumber first, then shifted to a higher wavenumber. Infrared spectrum character of collagen, nucleic acid and proteoglycan were compared and analyzed. The ratios of I1 657/I 1 548, I1 074/I1 548 and I1 074/I1 237 tend to decrease with ages increasing in surface and middle. However, the ratios at 2 months are less than other ages in deep. These results are consistent with the regular pattern of cartilage ingredient change in different degradation stage, while the tibial platform images created by microscopic spectral imaging technology is highly compliant with pathology description. The authors' primary result illustrated that FTIR microspectroscopy can be used for in-situ analysis of molecular constituents of different levels cartilages. The molecular information obtained from the study is important for understanding the pathogenesis of cartilage diseases.

Pranoprofen inhibits endoplasmic reticulum stress-mediated apoptosis of chondrocytes.

BACKGROUND: Endoplasmic reticulum stress (ERS) is a newly discovered pathway that causes apoptosis. At present, there is little research about the non-steroidal anti-inflammatory drug (NSAID) on the apoptosis of chondrocytes CHs.Our study aimed to test the anti-apoptosis effects of pranoprofen (PF), a specific prostaglandin E2 (PGE2) inhibitor, on human CHs. METHODS: We firstly made a distinguish about the levels of PGE2, ERS, andapoptosis between cartilage with and without OA. Then CHs isolated from healthy cartilage were pretreated H2O2 or Tunicamycin (TM) to activate ERS, and then exposed to PF. Expression of type II collagen, Runx-2, COX-9, SOD1, GPX1, GRP78, CHOP, caspase-12, ROS level, and apoptosis cell ratio was determined by immunofluorescence, Western blot, RT-PCR, or flow cytometry respectively. RESULTS: We found that oxidative stress, PGE2, ERS, and apoptosis were upregulated in OA cartilage. In addition, H2O2 and TM could increase the levels of PGE2, GRP78, CHOP, caspase-12, and reactive oxygen species (ROS), resulting in the degeneration of CHs. PF significantly reduced the expression of PGE2 and suppressed the ERS and apoptosis caused by H2O2 and TM, showing a protective function of CHs degeneration. CONCLUSIONS: This study demonstrates the anti-apoptotic effect of PF in the abrogation of the ERS-mediated apoptosis in human CHs, suggesting a new mechanism of PF in the treatment of OA.

Poly[[triaqua-(µ(3)-pyridine-2,4,6-tri-car-boxyl-ato)gadolinium(III)] monohydrate].

The title compound, {[Gd(C(8)H(2)NO(6))(H(2)O)(3)]·H(2)O}(n), was obtained in water under hydro-thermal conditions. The Gd(III) ions are nine-coordinated by two O and one N atoms from one pyridine-2,4,6-tricarboxyl-ate ligand, two O atoms from another ligand, one O atom from a third ligand and three coordinated water mol-ecules. Each ligand binds three metal centers. Two-dimensional layers are formed through the Gd-O bonds and the layers are linked by O-H⋯O hydrogen bonds, forming a three-dimensional network.

Effect of chengzai pill on the L-type Ca2+ channel currents of osteoblasts pretreated with methylprednisolone.

OBJECTIVE: To observe the effect of the serum containing Chengzai Pill on the L-type voltage-sensitive calcium channels current (L-VSCCsC) of osteoblastic MC3T3-E1 cells pretreated with methylprednisolone (mPSL). METHODS: A control group, a model group, a low dose group and a high dose group were set up. The whole cell patch clamp technique was used to record L-VSCCsC of 10 osteoblastic MC3T3-E1 cells in each group and their peak currents were determined. RESULTS: The peak current of the control group was 0.2284 +/- 0.0209 nA; the peak current of the model group was 0.1839 +/- 0.0179 nA; decreased by 19.5% as compared with the control group (P<0.01); the peak current of the low and high dose groups was 0.2526 +/- 0.0093 nA and 0.2671 +/- 0.0120 nA respectively, increased by 37.4% and 45.2% as compared with the model group (P<0.01); the difference between the low and high dose groups was P<0.05. CONCLUSION: 1. mPSL inhibits L-VSCCsC of osteoblasts; and 2. The serum containing Chengzai Pill increases L-VSCCsC of osteoblasts pretreated with mPSL.

[Preliminary study of intervention in effect of Bushen Huoxue recipe on calcification of lumbar vertebra cartilage endplate of the aging gerbils].

OBJECTIVE: To study the effect of Bushen Huoxue decoction on calcification of cartilage endplate in lumbar vertebrae. METHODS: Six healthy male gerbils with 2-month-old were selected as normal control group, and 24 7-month-old healthy male gerbils were fed to 12-month-old to establish the aged gerbil model. Thirty gerbils were randomly divided into five groups as follow: the normal control group (n=6), model group (n=6, normal saline 4 ml/kg, intragastric 30 d), Bushen Huoxue low dose group(n=6, 1.9× 10⁻ ³ ml/g given Bushen Huoxue recipe orally, 30 d), Bushen Huoxue middle dose group(n=6, 3.8× 10⁻ ³ ml/g given Bushen Huoxue recipe orally, 30 d), Bushen Huoxue high dose group(n=6, 7.6× 10⁻ ³ ml/g given Bushen Huoxue recipe orally, 30 d), the intervention group administered for 1.36 g from 7-month-old age, 30 d. The animals were sacrificed at the age of 2 months in the normal control group and 12 months of age in the other groups. The morphology of the lumbar vertebral cartilage endplate, the area of vascular bud, the ratio of non-calcified/calcified layer were analysis by HE chromosome visual method. The expression of type X collagen and BMPs in cartilage endplates were detected by rabbit monoclonal immunohistochemical staining. RESULTS: The relative area of the vascular buds cartilage endplate measurements showed that compared with the model group, middle dose group and normal control group increased (P<0.05), high and low dose groups all had different degrees of increase, but no statistical significance(P>0.05). The ratio of cartilage endplate thickness of non-calcified/calcified showed that compared with the model group, Bushen Huoxue middle dose, normal control group increased, with statistical significance(P<0.05), and high and low dose groups all had different degrees of increase, but there were no statistical significance(P>0.05). Compared with the model group, the expression of type X collagen in the cartilage endplate of the normal group, the Bushen Huoxue low, middle and high dose groups decreased, and had statistical significance(P<0.01); compared with the model group, the expression of BMPs in the normal group, Bushen Huoxue middle dose group increased, with statistically significant(P<0.01), while the high and low dose groups increased in different degrees, but there was no statistical significance(P>0.05). CONCLUSIONS: Bushen Huoxue prescription can delay the calcification of cartilage endplate in the process of aging, suggesting that it can be used as a preventive medicine for early disc degeneration.

Bone-strengthening pill (BSP) promotes bone cell and chondrocyte repair, and the clinical and experimental study of BSP in the treatment of osteonecrosis of the femoral head.

About 1 in 3 people suffer from bone and joint disease, which is a disease of bone and cartilage cells. Osteonecrosis of the femoral head (ONFH) is a typical example of bone and joint disease involving bone cell necrosis. Osteonecrosis of the femoral head leads to the occurrence of premature osteoarthritis of the hip and collapse of the cartilage cells, and there is currently no effective drug treatment available. In order to study the effects of "bone-strengthening pill" (BSP) on the repair of bone and cartilage cells, we investigated the potential effects of the herbal mixture BSP in an animal model of avascular necrosis of the femoral head and in patients. Results showed that 90% of rats injected with prednisone developed ONFH, whereas BSP administration prevented ONFH development in 70% of prednisone-injected rats. We evaluated the constituents of BSP by HPLC fingerprinting. We also evaluated the clinical efficacy of BSP in a double-blind, randomized, controlled trial of 300 patients with ONFH. The response rate was found to be higher in the treatment group than in the control group, with a response rate of 82% in the treatment group. Treatment with BSP also significantly reduced pain, improved hip function, reduced lameness, and improved pathology by X-ray and MRI analysis, compared with patients who did not receive BSP. These results suggest that BSP treatment inhibits and reverses necrosis of the femoral head bone cells and cartilage cells to repair the femoral head, promote the repair of bone and cartilage diseases.

[Analysis of clinical characteristics and image for sternum tumor].

OBJECTIVE: To analyze the clinical characteristics and imaging features of sternum tumor, in order to improve the knowledge of the diseases. METHODS: The clinical data and imaging data of 18 patients with sternum tumor were retrospectively analyzed. There were 13 males and 5 females, aged from 24 to 73 years old with an average of 58.3 years. Eleven patients were treated by conservative treatment and 7 patients were treated by surgery. The course of disease ranged from 2 months to 6 years. One patient had no obvious symptoms and 1 patient had a history of chest trauma. Lesion location, pattern of bone destruction, cortical bone abnormality, MRI signal characteristics and contrast enhancement were observed in the patients. RESULTS: Eighteen cases included malignant bone tumor in 16 cases and benign tumor in 2 cases;and 7 cases located in sternal manubrium, 9 cases located in sternal body, and 2 cases located in both. On the imaging, sternal tumor had expansively developing tendency, and the bone destruction zone boundary continuity or partial disruption occurred in 15 cases, the border of cortical bone thinning in 2 cases, the cortical bone increased thickness in 1 case. No bone marrow edema around tumor was found in 7 cases by MRI and soft tissues was encroached in 2 cases; in T1WI of MRI, 4 cases showed the hypointense signal and 3 cases showed the iso or hyper intensity signal; in T2WI of MRI, 6 cases showed the iso or hyper intensity signal and 1 case showed confounding signal. Enhancement scanning in MRI, premodinantly tumors showed obviously strengthening. CONCLUSIONS: The types of the sternum tumors are various and the most tumors are malignant. The images of the mostly sternum tumor absented same charactiristic, and it must combine with clinical manifestation to analyze the X rays, CT scanning and MRI, it will conduce to improve the diagonstic accuracy of the sternum tumors.

Experimental study on cheng zai wan for treatment of necrosis of the femoral head.

Cheng Zai Wan ([symbol: see text]), a Chinese herbal preparation was administrated in the two-leg rat model of aseptic necrosis of the femoral head established by taking prednisone acetate for a long period and the osteoporosis model rat by castration in order to explore the effects of the prescription on necrosis of the femoral head. The results showed that after treatment, the pitting on the surface of the femoral head disappeared, the reticular structure with filling cells was restored; the fat droplets in bone cells or cartilage cells of the femoral head were significantly reduced; sparse capillaries were improved, density and width of the bone trabecula were increased somewhat; bone mineral density, bone weight, bone strength and rigidity were significantly increased; and the low level of estrin was improved. It is suggested that Cheng Zai Wan has definite therapeutic effects on aseptic necrosis of the femoral head.

[Preliminary investigation of animal model of tibial defection fracture made by drilling hole method ].

OBJECTIVE: To compare the differences in damage degree, healing time and healing process between two animal models of defection fracture and provide the research data for the establishment of a new animal model. METHODS: Sixty male SPF Spragur-Dawley rats were divided randomly into control group, sawing group and drilling hole group, with 20 rats in each group. Animals of drilling hole group were treated with hone drill and made an 1 mm hole on tibial plateau animals of sawing group were treated with saw and made an 3 mm defection on tibial plateau; nothing to do in animals of control group. The rats were respectively killed at 2nd, 4th weeks after operation, bone density, bone ash quantity, contents of bone Ca, P were detected. RESULTS: At the 2nd week after operation, bone density, bone ash quantity, contents of bone Ca, P in drilling hole group and sawing group were significantly lower than that of the control group (P < 0.05 or P < 0.01). Two animal models of drilling hole group and sawing group were similar in X-ray films and pathology investigations. At the 4th week after operation, there were no significant defference in bone density and bone ash quantity among three groups (P > 0.05). Rats of sawing group showed more poroma and inflammatory infiltration in histological examination. CONCLUSION: Drilling hole method and sawing method could be used to make animal model of defection fracture, two methods showed similar damage degree, healing time and healing process, hut drilling hole method have advantages of simple operation, easily control damage degree and less inflammatory infiltration and bone disunion.

[Influence of aqueous extract of Aralia echinocaulis Hand.-Mazz on the expression of fracture healing-related factor receptors].

OBJECTIVE: To investigate the influence of aqueous extract of Aralia echinocaulis Hand.-Mazz on the expression of fracture healing-ralated factor receptors. METHODS: Single factor model was set up in SD rat. Selecting 14 and 28 days in the experiment. Immunohistochemistry was employed to determine the expression of Fibroblast growth factor receptor 2 (FGFR2), Fms-like tyrosine kinase (Flt-1) and Fetal licer kinase (Flk-1) at 14 and 28 days after model establishing. RESULTS: The expression of Flt-1 and Flk-1 at 14 days (the latter was more remarkable) were obviously promoted in High dose group of aqueous extract of Aralia echinocaulis Hand.-Mazz, and higher than that in normal group and model group. The expression of FGFR2 in the high dose group of Aralia echinocaulis Hand -Mazz was also promoted visibility, close to that in the compare group (traditional Chinese medicine), but higher than than in the model group. There was no significant difference among them. At 28 days, the expression of FGFR2, Flt-1 and Flk-1 in all groups decreased except normal group, and got higher expression in model groups than each control groups. CONCLUSION: Aqueous extract of Aralia echinocaulis Hand.-Mazz can promote angiogenesis in fracture healing, improve the activity and aggregation of fibroblasts, osteoblasts and chondrocytes. It also helps to quicken ossification in the cartilage and promote fracture healing.

[In vivo effects of Qizheng-xiaotong plaster on soft tissue injury of rabbit ears].

OBJECTIVE: To explore in vivo effects of Qizheng-xiaotong plaster on soft tissue injury in rabbit ears at different periods and to offer theoretical bases for clinical application. METHODS: The experimental models of soft tissue injury in ears were produced in 10 New Zealand white rabbits, and the ears were divided into three groups at the 1st, 2nd, 3rd week. The normal group and treatment group were given the Qizheng-xiaotong plaster extract, and the model group with normal saline. Microscopic analysis, digital collection system, infrared temperature tester and thickness tester were applied to determine the changes of soft tissue injury in local microcirculation and the temperature change after 0, 0.5, 3 and 5 hours, and swelling change at 1 to 5 days respectively. RESULTS: At the 3rd hour, blood velocity speeded up in normal group and model group, and it lasted for two hours in model group. As compared with model group, it slowed down to original level in treatment group at the 5th hour and the soft tissue swelling decreased from the 3rd to the 5th day as well. CONCLUSION: The application of Qizheng-xiaotong plaster is effective in preventing further soft tissue oedema and haematoma. It can make the soft tissue swelling decreased at chronic stage compared with that at acute stage.

[Application of ultrasonic microscopy in biomechanical properties measurement of cancellous bone].

OBJECTIVE: Ultrasound scanning can provide the information of acoustic impedance through the ultrasonic reflection theory. This study tries to introduce the application of the ultrasonic technique in predicting biomechanical properties of cancellous bone. METHODS: The rat femoral head embedded in plexiglass was used as the bone specimens for ultrasound scanning and the gray scale images scanned by ultrasound microscope were collected and analyzed. RESULTS: The value of acoustic impedance was calculated by analyzing the gray scale image. CONCLUSION: The application of ultrasonic microscopy can show more value about the microstructure of biomechanical properties of bone tissue.