Nerve growth factor(NGF) has a critical role in peripheral nerve regeneration. The aim of this study is to construct a well-functioning hNGF-$\beta$ recombinat adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with adenovirus mediated hNGF-$\beta$ gene transfection into Schwann cells. First PCR associated cloning of GFP-tagged hNGF-$\beta$ which was ligated into E1/E3 deleted adenoviral vector was performed and tranfected into E. coli to construct hNGF-$\beta$ recombinant adenovirus. After production of recombinat adenovirus in a large scale, its transfection efficiency, expression, and function were evaluated using cell lines or primarily cultured cells of HEK293 cells, Schwann cells, fibroblast(NIH3T3) and myocyte(CRH cells). GFP expression was observed in 90% of infected cells compared to uninfected cells. Total mRNA isolated from hNGF-$\beta$ recombinat adenoviru infected cells showed strong RT-PCR band, however, LacZ recombinant adenovirus infected or uninfected cells did not. NGF quantification by ELISA showed a maximal release of 18.865 +/- 0.31ng/mL at 4th day. PC-12 cells exposed to media with hNGF-$\beta$ recombinant adenovirus infected Schwann cell demonstrated higher levels of differentiation compared with controls. We generated hNGF-$\beta$ recombinant adenovirus and induced over expression of NGF successfully in nonneuronal and neuronal cells. Following these result, it is expected to develop an improved treatment strategy peripheral nerve regeneration using the hNGF-$\beta$ gene transfected cells.
Thromboelastography(TEG) enables a global assessment of hemostatic function to be made from a single blood sample, documenting the interaction of platelets with protein coagulation cascade from the time of the initial platelet-fibrin interaction, through platelet aggregation, clot strengthening and fibrin cross linking to eventual clot Iysis. Thirty-five patients(mean age 34$\pm$ 12) undergoing open heart surgery from April 1st, 1996 to August 31th, 1996 were investigated at preoperatively and immediate, one hour, and 24 hours after cessation of cardiopulmonary bypass using TEG. Comparisons were made between classic hematological indices and TEG data. There were statistically significant correlation between maximal amplitude(MA) and platelet count before CPB, activating clotting time(ACT) and TEG date(R time, K time and a angle) at 24-hour after CPB. The data on the predictive accuracy for postoperative bleeding at 24-hour after CPB, the TEG was significantly better than ACT(57%) or the coagulation profiles(43%) as a predictor of postoperative bleeding, with an accuracy rate of 100% (P=0.0043). In conclusion, TEG seems to be easy to use, clinically accurate, cost effective and provides data which can effectively manage a patient's hemostasis.
The purpose of this research was to establish the culture condition for dissociated submandibu -lar gland (SG) cells. After trypsin digestion of SG from 3-4 weeks old mice, dissociated cells were cultured in 1OO/o fetal bovine serum-Dulbecco's modified Eagle's medium (FBS-DME) or 0.5-2% low protein serum replacement-DME (LPSR-DME) on plastic surface to form monolayer. The effects of FBS, LPSR and hormones on the growth and function of cultured SG cells were examined. SG cells dissociated by enzyme were successfully cultured and were characterized as epithelial-like cells by light and electron microscope. The maximal DNA synthesis of cultured SG cells was achieved by DME containing 5-10% FBS. The same results were obtained when the effects of LPSR on cell proliferation were examined up to a LPSR concentration of 2%. SG cells cultured in 20/o LPSR-DME expressed a population doubling time of 42.5 hrs and a saturation density of 1.2 $\times$10 5cell/cm$^2$. Dihydrotestosterone (DHT) in medium did not influence on the DNA synthesis of the cultured SG cells, but stimulated protein synthesis of the SG cells. Thyroxine (T4) stimulated protein synthesis of the SCI cells markedly in a dose-dependent fashion. EGF secretion by the cultured SG cells increased significandy by DHT and or T4 trearment. This finding indicated that secretion of EGF by the SG cells was under the control of the hormones such as androgen and thyroid hormones. It seems to be that the culture condition described here can be used as a useful tool for further research on the SCI cells.
The primary purpose of a TKA is to restore normal knee function Therefore, ideally, a TKA should: (a) maintain the natural leverage of the knee joint muscles to ensure generating adequate knee muscle moments to accomplish daily tasks such as rising from climbing stairs; (b) provide adequate knee joint stability. A 16-channel MyoResearch XP EMG system was used to collect the differential input surface electromyography signals VM, VL, RF, BF, ST during climbing/descending stair tests. A Peak Motion Measurement System was used to collect the kinematic and kinetic data. AKIN-COM Ill isokinetic dynamometer was used for EMG of VM, VL, RF, BF and ST during maximal voluntary contraction. I Quadriceps EMG results for the VM of the passed 1year group limb demonstrated significant less RMS EMG than that of the passed 3year group limb $60^{\circ}-15^{\circ}$ of knee flexion(p<0.05). The VL of the passed 1year group limb also demonstrated significants less RMS EMG than that of the passed 3year group limb from $60^{\circ}-45^{\circ}$ of knee flexion(p<0.05). Similar to the VM and VL, the RF of the passed 1year group limb showed less RMS EMG than that of the passed 3year group limb from $60^{\circ}-30^{\circ}$ do knee flexion(p<0.05). Hamstring EMG results for the BF of the passed 1year group limb demonstrated less RMS EMG than that of the passed 3year group limb from $75^{\circ}-15^{\circ}$ of knee flexion(p<0.05). The passed 1year group limb tended to have less ADD displacement(p<0.071) than that of the passed 3year group limb. There was no significant difference of the ABD displacement between the passed 1year group and the passed 3year group limbs(p<0.73). The passed 3year group used compensatory adaptation movement strategies to compensate for the strength deficit of passed 3year group limbs. The passed 3year group limb also increased the quadriceps muscle activation level to produce more knee extension moment to compensate for the short quadriceps moment arm. The passe 3year group limb might have an unstable knee joint in the medio-Iateral direction during the climbing/descending by showing a tendency of more ADD displacement and greater hamming co-activation EMG than the passed 1year group limbs. The TKA design was not able to help the knee joint to produce adequate knee extension moment with less quadriceps muscle effort. I think that old man needs continuous exercise for muscle strength.
Propofol is known to cause vasorelaxation of several systemic vascular beds. However, its effect on the pulmonary vasculature remains controversial. In the present study, we investigated the effects of propofol on human pulmonary arteries obtained from patients who had undergone surgery. Arterial rings were mounted in a Multi-Myograph system for measurement of isometric forces. U46619 was used to induce sustained contraction of the intrapulmonary arteries, and propofol was then applied (in increments from $10-300{\mu}m$). Arteries denuded of endothelium, preincubated or not with indomethacin, were used to investigate the effects of propofol on isolated arteries. Propofol exhibited a bifunctional effect on isolated human pulmonary arteries contracted by U46619, evoking constriction at low concentrations ($10-100{\mu}m$) followed by secondary relaxation (at $100-300{\mu}m$). The extent of constriction induced by propofol was higher in an endothelium-denuded group than in an endothelium-intact group. Preincubation with indomethacin abolished constriction and potentiated relaxation. The maximal relaxation was greater in the endothelium-intact than the endothelium-denuded group. Propofol also suppressed $CaCl_2$-induced constriction in the 60 mM $K^+$-containing $Ca^{2+}$-free solution in a dose-dependent manner. Fluorescent imaging of $Ca^{2+}$ using fluo-4 showed that a 10 min incubation with propofol ($10-300{\mu}m$) inhibited the $Ca^{2+}$ influx into human pulmonary arterial smooth muscle cells induced by a 60 mM $K^+$-containing $Ca^{2+}$-free solution. In conclusion, propofol-induced arterial constriction appears to involve prostaglandin production by cyclooxygenase in pulmonary artery smooth muscle cells and the relaxation depends in part on endothelial function, principally on the inhibition of calcium influx through L-type voltage-operated calcium channels.
Kim, Ha-Won;Shim, Mi-Ja;Kim, Won-Bae;Kim, Byong-Kak
BMB Reports
/
v.28
no.6
/
pp.527-532
/
1995
Human taurine transporter has 12 transmembrane domains and its molecular weight is 69.6 kDa. The long cytoplasmic carboxy and amino termini might function as regulatory attachment sites for other proteins. Six potential protein kinase C phosphorylation sites have been reported in human taurine transporter. In this report, we studied the effects of phorbol 12-myristate 13-acetate (PMA) and glucocorticoid hormone on taurine transportation in the RAW 264.7, mouse macrophage cell line. When the cells were incubated with $[^{3}H]taurine$ in the presence or absence of $Na^+$ ion for 40 min at $37^{\circ}C$, the [$[^{3}H]taurine$ uptake rate was 780-times higher in the $Na^{+}-containing$ buffer than in the $Na^{+}-deficient$ buffer, indicating that this cell line expresses taurine transporter protein on the cell surface. THP1, a human promonocyte cell line, also showed a similar property. The $[^{3}H]taurine$ uptake rate was not influenced by the inflammatory inducing cytokines such as interleukin-1, gamma-interferon or interleukin-1+gamma-interferon, but was decreased by the PMA in the RAW 264.7 cell line. This suggests that activation of protein kinase C inhibits taurine transporter activity directly or indirectly. The inhibition of $[^{3}H]taurine$ uptake by PMA was time-dependent. Maximal inhibition occurred in one hr stimulation with PMA Increasing the treatment time beyond one h reduced the $[^{3}H]taurine$ uptake inhibition due to the depletion or inactivation of protein kinase C. The cell line also showed concentration-dependent $[^{3}H]taurine$ uptake under PMA stimulation. The phorbol-ester caused 23% inhibition at the concentration of 1 ${\mu}m$ PMA. The inhibition was significant even at a concentration as low as 10 nM PMA The reduced $[^{3}H]taurine$ uptake could be recovered by treatment with glucocorticosteroid hormone. Dexamethasone led to recover of the reduced taurine uptake induced by phorbol-ester, recovering maximally after one hr. This may suggest that macrophage cells require higher taurine concentration in a stressed state, for the secretion of glucocorticoid hormone is increased by hypothalamo-pituitary-adrenocortical (HPA) axis activation in the blood stream.
Lee, Yun Jung;Kho, Min Chul;Tan, Rui;Lee, Jae Yun;Hwang, Jin Seok;Cha, Jeong Dan;Choi, Kyung Min;Kang, Dae Gill
The Korea Journal of Herbology
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v.30
no.6
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pp.69-75
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2015
Objectives : This study was designed to investigate effects of the combination with Korean Red Ginseng (Panax ginseng C.A. Meyer), Gastrodia Rhizoma (Gastrodia elata Blume) and Polygoni Multiflori Radix (Polygonum multiflorum Thunberg) on metabolic disorders including cholesterol and erectile dysfunction in hyperlipidemia rats.Methods : Animals were divided into six groups; Control with normal diet, high fat/cholesterol-diet (HFCD), fluvastatin, Korean Red Ginseng treated (KRG), and the combination treated (Korean Red Ginseng, Gastrodia Rhizoma and Polygoni Multiflori Radix; 1:1:1 for KGP1 and 2:1:1 for KGP2). The experimental groups initially received HFCD for 10 weeks and then treated orally with fluvastatin, KRG, KGP1 and KGP2 during the final 6 weeks. Erectile function was determined by the measurements of intracavernosal pressure (ICP) and maximal arterial pressure (MAP) after electrical stimulation of the cavernosal nerve.Results : KGP2 decreased the level of total cholesterol and LDL cholesterol in the sera of HFCD rats without no changes of body weights. KRG, KGP1 and KGP2 decreased the level of C-reactive protein (CRP) levels except of fluvastatin, synthetic HMG-CoA reductase inhibitor. KRG, KGP1 and KGP2 significantly increased the ICP, ICP/MAP ratio, area under the curve (AUC) compared with those of normal rat. Morphometric analyses showed that KRG, KGP1 and KGP2 increased the volume of smooth muscle and the regular arrangement of collagen fibers in corpus cavernosum of HFCD rats. The penile expression of eNOS was increased by KRG, KGP1 and KGP2.Conclusions : Based on these results, we suggest that the combination with Korean Red Ginseng, Gastrodia Rhizoma and Polygoni Multiflori may improve hyperlipidemia through regulating the lipid profiles and erectile dysfunction in rats.
Park, Hae-Kun;Kim, Kwang-Jin;Sung, Hae-Sook;Jeon, Byung-Sook
The Korean Journal of Physiology
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v.11
no.2
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pp.45-50
/
1977
The maximum voluntary ventilation (MVV) is one of the most widely used pulmonary function test, but its measuring method was very difficult and unreliable. However, it is need to get more easy and simple measuring method of MVV. Therefore, this study was attempted to get more easy and simple measuring method of MVV by means of the forced expiratory volume $(FEV_{T})$. The young and healthy 1,000 Korean students(592 male and 408 female) were cheesed for this purpose and whose ages were from 8 to 20 years. A spirometer (9L, Collins Co.) was used for the MVV and FEV, and they were measured 3 times at standing position, and the highest value was used. In the measurements, the subjects for MVV were asked for the breath as fast and deeply as possible for 12 seconds, and for FEV were asked for the rapid and forceful exhalation after a maximal inhalation (forced expiratory curve). In the FEV measurements toward the end of the expiration, the subjects were exhaused to continue the effort until no further gas was expired. During these measurements, the investigator stood by the subject to give a constant encouragement. FEV were calculated in the volume exhaled during the one-half $(FEV_{0{\cdot}5,}\;ml)$, the first second $(FEV_{1{\cdot}0,}\;ml)$ and the percentage of the total vital capacity exhaled during the one-half second $(FEV_{0{\cdot}5,}\;%)$. The results are summarized as follows: 1) The values of MVV were increased linearly with ages until 20 in both sexes. The values of male at the age of 20 was $168.2{\pm}2.5L/min$, and female at the age of 17 was $112.3{\pm}3.0L/min$, respectively. 2) The values of FEV (ml) were increased linearly with ages until 20 in both sexes. The values of $FEV_{0{\cdot}5}$ were $2,797{\pm}65.7ml$ in the male of 20 years and were $2,088{\pm}54.6ml$ in the female of 17 years, and of $FEV_{1{\cdot}0$ were $4,119{\pm}68.2ml$ in the male of 20 years and were $2,897{\pm}65.9ml$ in the female of 17 years, respectively. 3) The correlation coefficients between MVV and $FEV_{0{\cdot}5}\;or\;FEV_{1{\cdot}0$ (ml) were 0.82 or 0.85 in the male, and 0.77 or 0.79 in the female, respectively. 4) The prediction formulae for MVV to be derived from above results were: For male: MVV (L/min) =7.19+$0.05{\times}FEV_{0\cdot5}(ml)$, MVV (L/min)=11.25+$0.04{\times}FEV_{1\cdot0}(ml)$ For female: MVV (L/min)=16.03+$0.05{\times}FEV_{0\cdot5}(ml)$, MVV (L/min)=9.47+$0.03{\times}FEV_{1\cdot0}(ml)$.
Journal of the Korea Academia-Industrial cooperation Society
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v.12
no.12
/
pp.5766-5772
/
2011
This study was to investigate the change of gait temporal parameter and ankle joint moment between patients with achilles tendinitis and healthy people. Thus, the purpose of this study is to clarify biomechanical change of gait in patients with achilles tendinitis and to find risk factor for prevention of achilles tendinitis. We recruited 20 patients with an achilles tendinitis and 20 healthy people. While subjects shuttled 5 times on 13 m distance with comfortable pace, we examined gait function marker with three-dimensional gait analysis system. All subject outstepped center of forceplate during gait and calculated ankle joint moment using software. Obtained data was analyzed using SPSS 12.0 software. In results, we confirmed that patients with achilles tendinitis showed reduction of extension moment in early initial phase and reduction of flexion moment in mid-stance on hip joint. and reduction of flexion moment in early initial phase and reduction of extension moment in late phase on knee joint. And we identified that patients with achilles tendinitis showed reduction of dorsiflexion moment in early stance phase, maximal plantarflexion moment in mid stance phase, and dorsiflexion moment in late stance phase. Thus, there are biomechanical changes on gait in patients with achilles tendinitis compared to healthy people. And, in clinical settings, they should focus on changes of gait in patients with achilles tendinitis. Further study will be undertaken for the biomechanical changes of patietns with achilles tendinitis.
Lee, Subum;Roh, Sung Woo;Jeon, Sang Ryong;Park, Jin Hoon;Kim, Kyoung-Tae;Lee, Young-Seok;Cho, Dae-Chul
Journal of Korean Neurosurgical Society
/
v.64
no.5
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pp.791-798
/
2021
Objective : The period of mechanical ventilator (MV)-dependent respiratory failure after cervical spinal cord injury (CSCI) varies from patient to patient. This study aimed to identify predictors of MV at hospital discharge (MVDC) due to prolonged respiratory failure among patients with MV after CSCI. Methods : Two hundred forty-three patients with CSCI were admitted to our institution between May 2006 and April 2018. Their medical records and radiographic data were retrospectively reviewed. Level and completeness of injury were defined according to the American Spinal Injury Association (ASIA) standards. Respiratory failure was defined as the requirement for definitive airway and assistance of MV. We also evaluated magnetic resonance imaging characteristics of the cervical spine. These characteristics included : maximum canal compromise (MCC); intramedullary hematoma or cord transection; and integrity of the disco-ligamentous complex for assessment of the Subaxial Cervical Spine Injury Classification (SLIC) scoring. The inclusion criteria were patients with CSCI who underwent decompression surgery within 48 hours after trauma with respiratory failure during hospital stay. Patients with Glasgow coma scale 12 or lower, major fatal trauma of vital organs, or stroke caused by vertebral artery injury were excluded from the study. Results : Out of 243 patients with CSCI, 30 required MV during their hospital stay, and 27 met the inclusion criteria. Among them, 48.1% (13/27) of patients had MVDC with greater than 30 days MV or death caused by aspiration pneumonia. In total, 51.9% (14/27) of patients could be weaned from MV during 30 days or less of hospital stay (MV days : MVDC 38.23±20.79 vs. MV weaning, 13.57±8.40; p<0.001). Vital signs at hospital arrival, smoking, the American Society of Anesthesiologists classification, Associated injury with Injury Severity Score, SLIC score, and length of cord edema did not differ between the MVDC and MV weaning groups. The ASIA impairment scale, level of injury within C3 to C6, and MCC significantly affected MVDC. The MCC significantly correlated with MVDC, and the optimal cutoff value was 51.40%, with 76.9% sensitivity and 78.6% specificity. In multivariate logistic regression analysis, MCC >51.4% was a significant risk factor for MVDC (odds ratio, 7.574; p=0.039). Conclusion : As a method of predicting which patients would be able to undergo weaning from MV early, the MCC is a valid factor. If the MCC exceeds 51.4%, prognosis of respiratory function becomes poor and the probability of MVDC is increased.
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