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Comparative metabolomic analysis in horses and functional analysis of branched chain (alpha) keto acid dehydrogenase complex in equine myoblasts under exercise stress

  • Jeong-Woong, Park;Kyoung Hwan, Kim;Sujung, Kim;Jae-rung, So;Byung-Wook, Cho;Ki-Duk, Song
    • Journal of Animal Science and Technology
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    • v.64 no.4
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    • pp.800-811
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    • 2022
  • The integration of metabolomics and transcriptomics may elucidate the correlation between the genotypic and phenotypic patterns in organisms. In equine physiology, various metabolite levels vary during exercise, which may be correlated with a modified gene expression pattern of related genes. Integrated metabolomic and transcriptomic studies in horses have not been conducted to date. The objective of this study was to detect the effect of moderate exercise on the metabolomic and transcriptomic levels in horses. In this study, using nuclear magnetic resonance (NMR) spectroscopy, we analyzed the concentrations of metabolites in muscle and plasma; we also determined the gene expression patterns of branched chain (alpha) keto acid dehydrogenase kinase complex (BCKDK), which encodes the key regulatory enzymes in branched-chain amino acid (BCAA) catabolism, in two breeds of horses, Thoroughbred and Jeju, at different time intervals. The concentrations of metabolites in muscle and plasma were measured by 1H NMR (nuclear magnetic resonance) spectroscopy, and the relative metabolite levels before and after exercise in the two samples were compared. Subsequently, multivariate data analysis based on the metabolic profiles was performed using orthogonal partial least square discriminant analysis (OPLS-DA), and variable important plots and t-test were used for basic statistical analysis. The stress-induced expression patterns of BCKDK genes in horse muscle-derived cells were examined using quantitative reverse transcription polymerase chain reaction (qPCR) to gain insight into the role of transcript in response to exercise stress. In this study, we found higher concentrations of aspartate, leucine, isoleucine, and lysine in the skeletal muscle of Jeju horses than in Thoroughbred horses. In plasma, compared with Jeju horses, Thoroughbred horses had higher levels of alanine and methionine before exercise; whereas post-exercise, lysine levels were increased. Gene expression analysis revealed a decreased expression level of BCKDK in the post-exercise period in Thoroughbred horses.

Genotoxicity and Cytotoxicity in Human Cancer and Normal Cell Lines of the Extracts of Rhododendron brachycarpum D. Don leaves (만병초 잎 추출물의 유전 독성과 사람의 암세포주 등에 대한 세포독성)

  • Byun, Kyoung-Sup;Lee, Young-Woo;Jin, Hyou-Ju;Lee, Mi-Kyoung;Lee, Hyeon-Yong;Lee, Kun-Jae;Heo, Moon-Young;Yu, Chang-Yeon;Lee, Jin-Ha
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.4
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    • pp.199-205
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    • 2005
  • This study was carried out to investigate the effect of 70% ethanol extract and each fraction from Rhododendron brachycarpum D. Don leaves on cytotoxicity, anticancer, genotoxicity and immunological activity in vitro bioassay. Cytotoxicity for human normal cells (HEL299 and Chang) of the samples was shown below 35% in 0.5 mg/ml concentration of samples except aqueous fraction by SRB assay. DNA damage on the Chang cell of the samples alone in comet assay was observed very weak damage activity even in high concentration (1 mg/ml) of the samples. The anticancer effect of the samples on human cancer cell lines (A549, AGS, Hep3B, MCF7) was indicated that the cancer cells were inhibited gradually in proportion to the increase of the concentration of the samples by MTT assay. The growth of the Raji and Jurkat cells were hastened by adding butanol fraction among the samples. In the genotoxicity on $H_2O_2-induced$ DNA damage in Chang cells using alkaline comet assay, most of samples were shown a strong protective activity from DNA OTM values.

Expression of steroidogenic enzymes in human placenta according to the gestational age

  • SO‑HYE HONG;SEUNG CHUL KIM;MEE‑NA PARK;JEA SIC JEONG;SEUNG YUN YANG;YOUNG JOO LEE;OK‑NAM BAE;HOE‑SAENG YANG;SUNGBAEK SEO;KYU‑SUP LEE;BEUM‑SOO AN
    • Molecular Medicine Reports
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    • v.19 no.5
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    • pp.3903-3911
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    • 2019
  • Female sex steroid hormones, including estradiol (E2) and progesterone (P4), serve significant physiological roles in pregnancy. In particular, E2 and P4 influence placenta formation, maintain pregnancy and stimulate milk production. These hormones are produced by ovaries, adrenal glands and the placenta, of which the latter is a major endocrine organ during pregnancy. However, the mechanism of hormone production during pregnancy remains unclear. In the present study, the regulation of steroid hormones and steroidogenic enzymes was examined in human placenta according to gestational age. In human placental tissues, expression levels of steroidogenic enzymes were determined with reverse transcription-quantitative polymerase chain reaction and western blotting. The mRNA and protein expression of CYP17A1, HSD17B3 and CYP19A1, which are associated with the synthesis of dehydroepiandrosterone (DHEA) and E2, was elevated at different gestational ages in human placenta. In addition, to evaluate the correlation between serum and placental-produced hormones, steroid hormone levels, including pregnenolone (PG), DHEA, P4, testosterone (T) and E2, were examined in serum and placenta. Serum and placenta expression of DHEA and E2 increased with gestational age, whereas T and P4 were differently regulated in placenta and serum. To confirm the mechanism of steroidogenesis in vitro, placental BeWo cells were treated with E2 and P4, which are the most important hormones during pregnancy. The mRNA and protein expression of steroidogenic enzymes was significantly altered by E2 in vitro. These results demonstrated that concentration of steroid hormones was differently regulated by steroidogenic enzymes in the placenta depending on the type of the hormones, which may be critical to maintain pregnancy.

Overexpression of the MUC1 Gene in Iranian Women with Breast Cancer Micrometastasis

  • Mansouri, Neda;Movafagh, Abolfazl;Soleimani, Shahrzad;Taheri, Mohammad;Hashemi, Mehrdad;Pour, Atefeh Heidary;Shargh, Shohreh Alizadeh;Mosavi-Jarahi, Alireza;Sasaninejad, Zahra;Zham, Hanieh;Hajian, Parastoo;Moradi, Hossein Allah;Mirzaei, Hamid Reza;Fardmanesh, Hedieh;Ohadi, Mina
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.sup3
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    • pp.275-278
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    • 2016
  • The membrane epithelial mucin MUC1 is expressed at the luminal surface of most simple epithelial cells, but expression is greatly increased in most breast cancers. The aims of present study were to investigate expression of the MUC1 gene and interactive affects in metastases. Whole cell RNA isolation from 50 sentinel lymph nodes (SNLs) of breast cancer patients was performed using reverse transcription and real-time PCR. All patients were diagnosed with breast cancer and without metastasis, confirmed by IHC staining. The evaluation of tumor and normal samples for expression of MUC1 gene, the results were 49.1% non-expressive and 45.3% expression (Student t, p = 0.03). Also in comparison of normal breast tissue and breast cancer SLN for MUC1 gene, MUC1 negative SLNs were 75.0% (18 samples) and MUC1 positive samples were 25.0% (6 samples). Over-expression of MUC1 gene may offer a target for therapy related to progression and metastasis in women with breast cancer.

Study on the Evaluation of Renal Function According to Set a Partial Region of Interest in 99mTc-DMSA scan of the Pediatric Patient with a Duplicated Ureter (중복요관을 가진 소아환자의 99mTc-DMSA 검사에서 부분적 관심영역 설정에 따른 신기능 평가에 관한 연구)

  • Nam-Koong, Hyuk;Oh, Shin Hyun;Kim, Jung Yul;Choi, Yoon Jung;Kim, Jae Sam;Lee, Chang Ho
    • The Korean Journal of Nuclear Medicine Technology
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    • v.17 no.1
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    • pp.43-47
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    • 2013
  • Purpose: A duplicated ureter is congenital renal malformations with ureter in two. Patients with duplicated ureter are in force to $^{99m}Tc-DMSA$ scan at surgery before and after. In existing examination, at produce result after $^{99m}Tc-DMSA$ scan, didn't compare to upper pole and lower pole with malformed kidney and compared to only relative uptake ratio. Therefore, this study will examine about utility of set a partial region of interest and to functional recovery of renal cell through change of upper pole uptake ratio of malformed kidney by setting each partial region of interest in upper pole and lower pole of malformed kidney in $^{99m}Tc-DMSA$ examination in surgery before and after. Materials and Methods: Pediatric patients with malformed kidney of incomplete duplicated ureter, 15 patients were enrolled in this study. Scanning were scan 3 to 4 hours after injection of $^{99m}Tc-DMSA$ 1.5 ~ 1.9 MBq/kg. Region of interest were each set in normal kidney, upper pole and lower pole with malformed kidney. Region of interest were set with same condition and method to images of surgery before and after that radio technologist 1 person, resident of nuclear medicine 1 person and doctor of urology together. Therefore, this study were compared to uptake ratio (A: B: C) that normal kidney (A), lower pole of malformed kidney (B) and upper pole of malformed kidney (C) about uptake ratio changes of malformed kidney in follow-up examination of surgery before and after. Results: When compared to 15 patients, uptake ratios were increased 7 persons and decreased 8 persons. Among increased 7 persons, it were periods of follow-up examination that 2 persons were 14 months, 4 persons were 12 months and 1 person was 8 months after surgery. Among decreased 8 persons, it were periods of follow-up examination that 4 persons were 12 months 3 persons were 6 months and 1 persons were 4 months after surgery. Conclusion: Existing study could not see the exact uptake ratio changes of malformed kidney because using only the overall Left-Right kidney uptake ratios. But a setting partial region of interest was able to see exactly what changes in the uptake of each upper pole and lower pole of malformed kidney. Because recovery of renal parenchymal cells is difficult in an evaluation of short period of time, follow-up examination should be made in long period of time. How to set up partial region of interest be thought that it would be useful.

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Relation of Interleukin-10 in Bronchoalveolar Lavage Fluid and Airway Inflammation in Bronchial Asthma (기관지천식에서 기관지폐포세척액내 IL-10과 기도염증정도의 연관성)

  • Lee, Sook-Young;Youn, Hung-Gue;Shin, Youn;Lee, Sang-Haak;Kim, Seok-Chan;Kim, Kan-Hyoung;Moon, Hwa-Sik;Song, Jeong-Sup;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.1
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    • pp.44-52
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    • 1999
  • Background : Airway infiltration by inflammatory cells, particularly of eosinophils, is one of the characteristic features of asthma. Several mechanisms for the recruitment of eosinophil is focused on the CD4+ T lymphocyte for the preferential production of Th2-c1erived cytokines. Interleukin-10(IL-10) is identified cytokine with potent antiinflammatory activity. This molecule has been shown to inhibit the release of cytokine from inflammatory cells including Th2 cell, and also to inhibit eosinophil survival. We therefore attempted to determine whether decreased synthesis of IL-10 in the lung of bronchial asthma may contribute to inflammation that is characteristics of this dease. Method: Subjects were patients with bronchial asthma(n=23) and normal controls(n=11). IL-10 produced from peripheral mononuclear cell(PBMC) and in bronchoalveolar lavage(BAL) fluid was measured by ELISA method. Degree of bronchial inflammation was assessed by total cell counts and eosinophil percents in BAL fluid, eosinophil infiltration on bronchial biopsy tissue and $PC_{20}$ for methacholine. Results: The IL-10 level produced by PBMC and in BAL fluid from patient with bronchial asthma were not different with normal controls(respectively, $901.6\pm220.4$ pg/ml, $810.9\pm290.8$ pg/ml for PBMC, $24.5\pm9.5$ pg/mL $30.5\pm13.5$ pg/ml for BAL fluid p>0.05). There were significant negative correlation between IL-10 in BAL fluid and eosinophil percents in BAL fluid or degree of eosinophil infiltration in bronchial biopsy (respectively r=-0.522, r=-0.4486 p<0.05). However there was no difference of IL-10 level according to $PC_{20}$ for methacholine. There were no correlation between IL-10 production by PBMC and peripheral blood eosinophil counts or serum eosinophilic cationic protein levels(respectively r=0.1146, r=0.0769 p>0.05). Conclusion: These observation suggest that IL-10 may participate but not acts the crucial role in regulation of the airway inflammation in bronchial asthma.

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Interleukin Levels in the Bronchoalveolar Lavage Fluid of Patients with Pulmonary Sarcoidosis (유육종증환자의 기관지폐포세척액내 Interleukin에 대한 연구)

  • Song, Jeong-Sup;Ahn, Joong-Hyun;Kim, Chi-Hong;Kim, Kwan-Hyoung;Moon, Hwa-Sik;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.5
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    • pp.1047-1057
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    • 1998
  • Background : Sarcoidosis is a systemic granulomatous disorder of unknown origin and characterized by accumulation of T cells and macrophages. Various cytokines may play crucial roles in the activation of T cells and macrophages, and thereby in the formation of granulomas. However, little is known about the balance between proinflammatory cytokines and antiinflammatory cytokines in the development of sarcoid granulomas and disease activities. In the present study, we measured IL-6, IL-8 and IL-10 in the bronchoalveolar lavage fluid(BALF) from patients with pulmonary sarcoidosis to find out whether there is an imbalance between proinflammatory cytokines and antiinflammatory cytokines in the lung. Methods: Fourteen subjects with the diagnosis of sarcoidosis and six healthy volunteers were included. BALF was concentrated ten-fold by pressure ultrafiltration and each cytokine levels were measured by EUSA method. Active sarcoidosis was defined by major organ involvement or clinically progressive diseases. Results: The mean IL-6 levels in the BALF of the active sarcoidosis group were significantly increased than in controls or inactive sarcoidosis group(p<0.05). Meanwhile, the IL-8 levels were increased and IL-10 levels were decreased in the active sarcoidosis group than in controls or inactive sarcoidosis group without significance(p>0.05). In active pulmonary sarcoidosis patients, the IL-6 levels in BALF correlated with the BALF CD4/CD8 ratio(r=0.768, p<0.05) and IL-8 levels(r=0.564, p<0.05). Conclusions : The data presented showed that pro-inflammatory cytokine IL-6 is important in the pathogenesis of sarcoidosis and decreased tendency of anti-inflammatory cytokine IL-10 might also be involved in the development of granulomatous inflammation in sarcoidosis.

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Superoxide Generation by Blood Monocyte and Pulmonary Alveolar Macrophage in Patients with Pulmonary Tuberculosis (폐결핵환자의 폐포대식세포 및 말초혈액내 단구세포에서 분비하는 과산화음이온의 비교 관찰)

  • Song, Jeong-Sup;Lee, Suk-Young;Jang, Jie-Jung;Kim, Young-Kyoon;Kim, Kwan-Hyoung;Moon, Hwa-Sik;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.1
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    • pp.11-19
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    • 1994
  • Background: Mycobacterium tuberculosis is a facultative intracellular pathogen which persists and multiplies within macrophage. Competent cell mediated immunity by cooperation of both T lymphocyte and macrophage of the host is required to kill the Mycobacterium tuberculosis. But a precise understanding of the pathogenesis of tuberculosis infection in pulmonary alveolar macrophage has not been achived. Research on the macrophage's basic microbicidal mechanism has elucidated the importance of oxygen-dependent or oxygen-independent components. Oxygen dependent processing begins with the reduction of oxygen by NADPH oxidase and generation of superoxide. In this study, the oxidative metabolic status of blood monocyte and pulmonary alveolar macrophage in patients with active pulmonary tuberculosis was accessed and compared with that of healthy control subjects to know whether there was a basic difference in superoxide generation by mononuclear cells between two groups. Methods: Pulmonary alveolar macrophage was purified after performing BAL(bronchoalveolar lavage) through the bronchi of infected lesion by plastic adhesion method. Blood monocyte was purified by Ficoll-Hypaque method. Superoxide generation by blood monocyte and pulmonary alveolar macrophage was measured by ferricytochrome-C reduction method after either stimulated with PMA(phorbol myristate acerate) or non-stimulated states. We also measured the effect of pulmonary tuberculosis patient's serum on superoxide generation by monocyte. Results: 1) Generation of superoxide by alveolar macrophage obtained from patients with pulmonary tuberculosis was little higher than those of controls, and PMA enhanced the generation of 2) Generation of superoxide by blood monocyte obtained from patients with pulmonary tuberculosis was little higher than those of control(p>0.05), and PMA more enhanced the generation of superoxide in patientswith pulmonary tuberculosis than those in controls(p<0.02). 3) Patient's serum enhanced the generation of superoxide by blood monocyte obtained from patients with pulmonary tuberculosis and controls, but not in the case of PMA stimulated blood monocyte. Conclusion: The present study suggest that the phenomenon of M.tuberculosis escape the microbicidal action of macrophage was not result of suppressed superoxide generation by blood monocyte and pulmonary alveolar macrophage, rather there might be a factor to stimulate the generation of superoxide by blood monocyte in pulmonary tuberculosis patient serum, but the comparision with effect of control's serum on superoxide generation needs further elucidation.

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Splenocyte-mediated immune enhancing activity of Sargassum horneri extracts (괭생이 모자반 추출물의 비장세포 면역활성 증강 효과)

  • Kim, Dong-Sub;Sung, Nak-Yun;Han, In-Jun;Lee, Byung-Soo;Park, Sang-Yun;Nho, Eun Young;Eom, Ji;Kim, Geon;Kim, Kyung-Ah
    • Journal of Nutrition and Health
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    • v.52 no.6
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    • pp.515-528
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    • 2019
  • Purpose: This study examined the immunological activity and optimized the mixture conditions of Sargassum horneri (S. horneri) extracts in vitro and in vivo models. Methods: S. horneri was extracted using three different methods: hot water extraction (HWE), 50% ethanol extraction (EE), and supercritical fluid extraction (SFE). Splenocyte proliferation and cytokine production (Interleukin-2 and Interferon-γ) were measured using a WST-1 assay and enzyme-linked immunosorbent assay, respectively. The levels of nitric oxide and T cell activation production were measured using a Griess assay and flow cytometry, respectively. The natural killer (NK) cell activity was determined using an EZ-LDH kit. Results: Among the three different types of extracts, HWE showed the highest levels of splenocyte proliferation and cytokine production in vitro. In the animal model, three different types of extracts were administrated for 14 days (once/day) at 50 and 100 mg/kg body weight. HWE and SFE showed a high level of splenocyte proliferation and cytokine production in the with and without mitogen-treated groups, whereas EE administration did not induce the splenocyte activation. When RAW264.7 macrophage cells were treated with different mixtures (HWE with 5, 10, 15, 20% of SFE) to determine the optimal mixture ratio of HWE and SFE, the levels of nitric oxide and cytokine production increased strongly in the HWE with 5% and 10% of SFE containing group. In the animal model, HWE with 5% and 10% of SFE mixture administration increased the levels of splenocyte proliferation, cytokine production, and activated CD4+ cell population significantly, with the highest level observed in the HWE with 5% of SFE group. Moreover, the NK cell activity was increased significantly in the HWE with 5% of SFE mixture-treated group compared to the control group. Conclusion: The optimal mixture condition of S. horneri with immune-enhancing activity is the HWE with 5% of SFE mixture. These results confirmed that the extracts of S. horneri and its mixtures are potential candidate materials for immune enhancement.

A Clinical Study on Multiple Myeloma (다발성 골수종의 임상적 고찰)

  • Hwang, Hyeong-Ki;Lee, Choong-Ki;Hyun, Myung-Soo;Shim, Bong-Sup;Lee, Hyun-Woo
    • Journal of Yeungnam Medical Science
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    • v.8 no.2
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    • pp.106-113
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    • 1991
  • A clinical review of 31 cases of multiple myeloma which were diagnosed by criteria of the SWOG between May 1983 and February 1990 at Yeungnam University Hospital was done. The results were as followings : 1. The peak incidence was in 7th decade and male to female ratio was 1.8 : 1. 2. The most common presenting symptom at first diagnosis was bone pain (58%), but fever, dyspnea, dizziness and palpable mass were also noted. 3. The distribution of laboratory findings as following diagnostic criteria of Southwest oncology group(SWOG) : plasmacytoma on tissue biopsy was noted 6 cases, bone marrow plasmacytosis with more than 10% plasma cells was 22 cases, monoclonal globulin spike on serum electrophoresis was 24 cases, lytic bone lesions was observed 22 cases. 4. Initial clinical stages were classified as 2 cases in stage I, 3 cases in stage II, 26 cases in stage III(84%) 5. Immunoelectrophoresis revealed the distribution of IgG 64%, light chain 22%, IgA 10%. Kappa to Lambda ratio of 1.1 : 1. 6. Hematologic & biochemical fingins revealed anemia with <8.5% of hemoglobulin in 42%, hypercalcemia with < 10.6mg% of serum calcium in 22%, azotemia >2.0mg% of serum creatinine in 19%. 7. The multiple punched out lesion of bone x-ray examination were noticed skull(65%), rib(42%), L-spine(35%), pelvis(23%), T-spine(19%). The initial skeletal roentgenographic findings showed osteoporosis, osteolytic lesion and fracture in 55%, only osteolytic lesion in 23%, only osteoporosis in 10%. 8. Complications of multiple myeoloma, such as 10 cases of renal impairment, 8 cases of infection, 16 cases of compression fracture of spine were observed.

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