2.Fusion of human orphan G protein-coupled receptors GPR45, GPR85 or GPR174 with Gi1α and their expression in insect Sf9 cells
Ming-li, PENG ; Chun-guang, HAN ; Zhi-qing, GAO ; Qiong, WANG ; Yue, GAO ; Yong-xue, LIU
Bulletin of The Academy of Military Medical Sciences 2010;34(1):8-11
Objective To obtain the fusion genes of several human orphan G protein coupled receptors (oGPCRs) with Gi1α subtype of G protein and their expression system. Methods The whole open reading frames of GPR45, GPR85, GPR174 and Gilα were cloned by RT-PCR from HepG2 cDNA separately,and the corresponding fusion genes were amplified by overlap extension PCR. Then, the fusion genes-containing pBacmids were successfully constructed with the Bac-to-Bac baculovirus expression system indicated by specific transposition and virus recombination. The insect Sf9 cells were transfected with pBacmid-oGPCRs-Gi1α, and the supernatant containing recombinant virus was harvested. With the supernatant, insect Sf9 cells were infected under an optimized condition (MOI=5, infection time=72 h) and the fusion proteins were prepared and detected by Western blotting.Results The three fusion genes of GPCR45, GPR85 or GPR174 with Gi1α were obtained. The corresponding fusion proteins could be properly prepared in Sf9 cells.Conclusion Human oGPCRs could be fused with Gilα, and the fusion genes could be expressed using the Bac-to-Bac baculovirus expression system in insect Sf9 cells.
3.Effect of electro-acupuncture stimulation of Ximen (PC4) and Neiguan (PC6) on remifentanil-induced breakthrough pain following thoracal esophagectomy.
Yan-Hu, XIE ; Xiao-Qing, CHAI ; Yue-Lan, WANG ; Yan-Chun, GAO ; Jun, MA
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(4):569-74
The clinical analgesic effect of electro-acupuncture (EA) stimulation (EAS) on breakthrough pain induced by remifentanil in patients undergoing radical thoracic esophagectomy, and the mechanisms were assessed. Sixty patients (ASAIII) scheduled for elective radical esophagectomy were randomized into three groups: group A (control) receiving a general anesthesia only; group B (sham) given EA needles at PC4 (Ximen) and PC6 (Neiguan) but no stimulation; and group C (EAS) electrically given EAS of the ipsilateral PC4 and PC6 throughout the surgery. The EAS consisting of a disperse-dense wave with a low frequency of 2 Hz and a high frequency of 20 Hz, was performed 30 min prior to induction of general anesthesia and continued through the surgery. At the emergence, sufentanil infusion was given for postoperative analgesia with loading dose of 7.5 μg, followed by a continuous infusion of 2.25 μg/h. The patient self-administration of sufentanil was 0.75 μg with a lockout of 15 min as needed. Additional breakthrough pain was treated with dezocine (5 mg) intravenously at the patient's request. Blood samples were collected before (T1), 2 h (T2), 24 h (T3), and 48 h (T4) after operation to measure the plasma β-EP, PGE2, and 5-HT. The operative time, the total dose of sufentanil and the dose of self-administration, and the rescue doses of dezocine were recorded. Visual Analogue Scale (VAS) scores at 2, 12, 24 and 48 h postoperatively and the incidence of apnea and severe hypotension were recorded. The results showed that the gender, age, weight, operative time and remifentanil consumption were comparable among 3 groups. Patients in EAS group had the lowest VAS scores postoperatively among the three groups (P<0.05). The total dose of sufentanil was 115±6.0 μg in EAS group, significantly lower than that in control (134.3±5.9 μg) and sham (133.5±7.0 μg) groups. Similarly, the rescue dose of dezocine was the least in EAS group (P<0.05) among the three groups. Plasma β-EP levels in EAS group at T3 (176.90±45.73) and T4 (162.96±35.00 pg/mL) were significantly higher than those in control (132.33±36.75 and 128.79±41.24 pg/mL) and sham (136.56±45.80 and 129.85±36.14 pg/mL) groups, P<0.05 for all. EAS could decrease the release of PGE2. Plasma PGE2 levels in EAS group at T2 and T3 (41±5 and 40±5 pg/mL respectively) were significantly lower than those in control (64±5 and 62±7 pg/mL) and sham (66±6 and 62±6 pg/mL) groups. Plasma 5-HT levels in EAS group at T2 (133.66±40.85) and T3 (154.66±52.49 ng/mL) were significantly lower than those in control (168.33±56.94 and 225.28±82.03) and sham (164.54±47.53 and 217.74±76.45 ng/mL) groups. For intra-group comparison, plasma 5-HT and PGE2 levels in control and sham groups at T2 and T3, and β-EP in EAS group at T3 and T4 were significantly higher than those at T1 (P<0.05); PGE2 and 5-HT levels in EAS group showed no significant difference among the different time points (P>0.05). No apnea or severe hypotension was observed in any group. It was concluded that intraoperative ipsilateral EAS at PC4 and PC6 provides effective postoperative analgesia for patients undergoing radical esophagectomy with remifentanil anesthesia and significantly decrease requirement for parental narcotics. The underlying mechanism may be related to stimulation of the release of endogenous β-EP and inhibition of inflammatory mediators (5-HT and PGE2).
4.Effect of Postoperative Continuous Epidural Analgesia with Sufentanil in Different Concentrations and Combined with Bupivacaine on Pain after Thoracotomy
Chun WANG ; Yue SU ; Wan-ming GENG ; Hui ZHENG ; Wei LIU ; Guangkuo GAO ; Zhengfang HU
Chinese Journal of Rehabilitation Theory and Practice 2006;12(4):352-354
ObjectiveTo observe the effect and safety of continuous epidural analgesia with sufentanil in different concentrations combined with 0.125% bupivacaine on pain after thoracotomy.Methods30 patients with ASA grade Ⅱ~Ⅲ and underwent thoracotomy were randomly divided into 3 groups treated with 0.125% bupivacaine combined with sufentanil 0.25 μg/ml (group A), 0.50 μg/ml (group B) and 0.75 μg/ml (group C) respectively. Before operation starting, epidural puncture was performed at T7~T8 and a catheter was put in. After operation, continuous epidural analgesia was performed by connecting the catheter and a analgesic pump. Analgesia effect was evaluated by visual analogous score (VAS) at sixth, twelfth, twenty-fourth and forty-eighth hours after operation. Dosage of assistant drug and side effects such as calmness, nausea, vomiting, skin pruritus and respiratory inhibition were also recorded.ResultsVAS scores and dosage of assistant drug of group B and group C were not different, but they were all lower than that of group A (P<0.05). Scores of skin pruritus of group A and group B were lower than that of group C (P<0.05), but there was no significant difference between group A and group B. No respiratory inhibition occurred in patients of all three groups.ConclusionContinuous epidural analgesia of 0.50 μg/ml sufentanil combined with 0.125% bupivacaine is safe and effective for patients after thoracotomy.
5.Efficacy of one lung protective ventilation regimen based on quarsi-static pressure-volume curve in patients undergoing thoracic surgery
Wenjun LIU ; Hui ZHENG ; Yue SU ; Wanming GENG ; Wei LIU ; Guangkuo GAO ; Chun WANG ; Zhengfang HU ; Hongyan JIA ; Na WANG
Chinese Journal of Anesthesiology 2011;31(8):950-954
ObjectiveTo evaluate the efficacy of PEEP set according to pressure-volume (P-V) curve for one lung ventilation (OLV) in patients undergoing thoracic surgery.MethodsOne hundred and twenty ASA Ⅱ or Ⅲ patients of both sexes aged 20-60 yr weighing 40-80 kg undergoing lobectomy under general anesthesia were enrolled in this study.Double lumen tube was inserted.Correct positioning was verified by fiberoptic bronchoscopy.The patients were mechanically ventilated.P-V curve was determined at 3 min of two-lung ventilation (TLV).Lower inflection point (LIP) was measured and the pressure at LIP (PLIP) was recorded.The patients were randomly divided into 5 groups (n = 24 each):control group (group C) and 4 lung protective ventilation regimen groups ( groups P1-4 ).PEEP was set at 0 and VT was set at 10 ml/kg in group C.PEEP was set at 0 and VT was set at 6 ml/kg in group P1.PEEP was set at PLIP- 2 cm H2O and VT was set at 6 ml/kg in group P2.PEEP was set at PLIP and VT was set at 6 ml/kg in group P3.PEEP was set at PLIP + 2 cm H2O and VT was set at 6 ml/kg in group P4.Peak airway pressure (Ppeak),plateau airway pressure (Pplat),airway resistance (Raw) and lung compliance (CL ) were measured and recorded during OLV and TLV after a period of stabilization.Arterial blood samples were taken before induction of anesthesia and at 20 min of TLV and 20 min of OLV for blood gas analysis.Qs/Qt was calculated.Arterial blood samples were collected at the beginning and end of OLV for determination of plasma concentrations of IL-6 and TNF-α(by ELISA).ResultsCompared with group C,Ppeak and Pplat were significantly increased while Raw was decreased and plasma IL-6 concentration was significantly decreased at the end of OLV in group P4.PaCO2 was significantly higher during TLV and OLV in groups P1-4 than in group C.There was no significant difference in the parameters of respiratory mechanics,blood gases and plasma IL-6 and TNF-α concentrations among groups P1,2.3.Compared with groups P1,2,3,Ppeak and Pplat were significantly increased while plasma IL-6 concentration was decreased at the end of OLV in group P4.ConclusionMechanical ventilation with VT set at 6ml/kg and PEEP at PLIP + 2 cm H2 O provides best venfilatory efficacy for OLV in terms of oxygenation and inhibition of inflammatory response.
6.The effects of si-wu-tang on serum protein of blood deficient mice induced by radiation.
Zeng-chun MA ; Yue GAO ; Hong-ling TANG ; Sheng-qi WANG
China Journal of Chinese Materia Medica 2003;28(11):1050-1053
OBJECTIVETo study the effects of Si-Wu-Tang on serum protein of blood deficient mice b y proteomicstechnique and study the enriching and regulating blood mechanism of Si-Wu-Tang on mocular level.
METHODThe blood deficient mice was induced by using a single dose of 3.5 Gy radiation from a 60Cogamma source, and high resolution two-dimensional polyacryamide gel electrophoresis (2-DE), computer-assisted image analysis, and mass spectrometry were used to detect regulated protein by Si-Wu-Tang.
RESULT12 lower and 4 higher protein in sera could be recovered by Si-Wu-Tang, 4 protein might be DNA-dependent protein kinase catalytic subunit, Dystrophin, KIF13A, dystonin. They play a part in DNA double-stranded break repair, recombination and modulation of transcription, transportation of mannose-6-phosphate receptor, etc.
CONCLUSIONSi-Wu-Tang can regulate serum protein in blood deficient mice, resulting in improving hematopoiesis and lessening irradiated injury.
Animals ; Autoantigens ; blood ; Carrier Proteins ; Cytoskeletal Proteins ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Dystonin ; Dystrophin ; blood ; Female ; Kinesin ; blood ; Mice ; Mice, Inbred C57BL ; Nerve Tissue Proteins ; Non-Fibrillar Collagens ; blood ; Plants, Medicinal ; chemistry ; Radiation Injuries, Experimental ; blood ; Radiation-Protective Agents ; pharmacology ; Whole-Body Irradiation
7.Analysis of myocardial perfusion and metabolism in patients with hypertrophic obstructive cardiomyopathy undergoing percutaneous transluminal septal myocardial ablation by99 Tcm-MIBI and 18F-FDG images
Jian-song, YUAN ; Shu-bin, QIAO ; Yue-qin, TIAN ; Ping-ping, HAN ; Wan-chun, ZHANG ; Wei-xian, YANG ; Run-lin, GAO ; Ji-lin, CHEN ; Yue-jin, YANG
Chinese Journal of Nuclear Medicine 2010;30(3):176-179
Objective To evaluate the use of gated SPECT in patients with hypertrophic obstructive cardiomyopathy (HOCM) and the effects of percutaneous transluminal septal myocardial ablation (PTSMA) on myocardial perfusion.Methods 99 Tcm-methoxyisobutylisonitrile (MIBI) and 18F-fluorodeoxyglucose (FDG) images were performed in 31 HOCM patients before PFSMA and in 15 patients 3-7 d after PTSMA.The images in different left ventricular segments were analysed by using scores.Results In 99Tcm-MIBI images, uptake decreased at the septal regions in 12 HOCM patients (80.0%, 12/15) after PTSMA, 18F-FDG images also showed decreased uptake at the septal regions in 5 HOCM patients (33.3%, 5/15) after PTSMA.Conclusion 99Tcm-MIBI images might be an important method to evaluate PTSMA results, and 18 F-FDG images showed important value as reference.
8.Involvement of leukotrine B4 receptors in the inflammatory responses and immunological regulation in vitro.
Chun-guang HAN ; Huo-gao HUANG ; Ming HU ; Wen-yan LUO ; Yue GAO ; Qiong WANG ; Yong-xue LIU
Chinese Journal of Applied Physiology 2009;25(2):273-276
AIMBLT1 and BLT2 were both recently cloned and identified as two subtypes of leukotrine B4 (LTB4) receptors. With the usage of U-75302 and LY255283, the specific antagonists of BLT1 and BLT2 respectively, the involvement of BLT1 and BLT2 in the inflammatory and immunological responses was in vitro explored.
METHODS(1) To investigate inhibition of U-75302 and LY255283 on the proliferation of rat synovial cells, 3H-TdR incorporation into the cells was quantified. (2) Flow cytometric assay for interferon-gamma (IFN-gamma) and interleukine 4 (IL-4) profiles in CD4+ T lymphocytes from rat spleen was carried out to determine the ratio of Th1/Th2.
RESULTS(1) For inhibition on rat synovial cells proliferation, U-75302 exerted its effect only at a high concentration of 10 micromol/L and LY255283 at the concentrations of 10 micromol/L-10 micromol/L. (2) Both U-75302 and LY255283 could elevate the percentage of Th2, but could not influence that of Th1.
CONCLUSIONBLT1 and BLT2 were involved in the synovial cells proliferation change the ratio of Th1/Th2. Their meaning served as targets for prevention and treatment of infectious diseases should be emphasized.
Animals ; Cell Line ; Cell Proliferation ; drug effects ; Fatty Alcohols ; pharmacology ; Glycols ; pharmacology ; Inflammation ; immunology ; Male ; Rats ; Rats, Wistar ; Receptors, Leukotriene B4 ; antagonists & inhibitors ; physiology ; Synovial Membrane ; cytology ; immunology ; Tetrazoles ; pharmacology ; Th1-Th2 Balance
9.Effect of electro-acupuncture stimulation of Ximen (PC4) and Neiguan (PC6) on remifentanil-induced breakthrough pain following thoracal esophagectomy.
Yan-hu XIE ; Xiao-qing CHAI ; Yue-lan WANG ; Yan-chun GAO ; Jun MA
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(4):569-574
The clinical analgesic effect of electro-acupuncture (EA) stimulation (EAS) on breakthrough pain induced by remifentanil in patients undergoing radical thoracic esophagectomy, and the mechanisms were assessed. Sixty patients (ASAIII) scheduled for elective radical esophagectomy were randomized into three groups: group A (control) receiving a general anesthesia only; group B (sham) given EA needles at PC4 (Ximen) and PC6 (Neiguan) but no stimulation; and group C (EAS) electrically given EAS of the ipsilateral PC4 and PC6 throughout the surgery. The EAS consisting of a disperse-dense wave with a low frequency of 2 Hz and a high frequency of 20 Hz, was performed 30 min prior to induction of general anesthesia and continued through the surgery. At the emergence, sufentanil infusion was given for postoperative analgesia with loading dose of 7.5 μg, followed by a continuous infusion of 2.25 μg/h. The patient self-administration of sufentanil was 0.75 μg with a lockout of 15 min as needed. Additional breakthrough pain was treated with dezocine (5 mg) intravenously at the patient's request. Blood samples were collected before (T1), 2 h (T2), 24 h (T3), and 48 h (T4) after operation to measure the plasma β-EP, PGE2, and 5-HT. The operative time, the total dose of sufentanil and the dose of self-administration, and the rescue doses of dezocine were recorded. Visual Analogue Scale (VAS) scores at 2, 12, 24 and 48 h postoperatively and the incidence of apnea and severe hypotension were recorded. The results showed that the gender, age, weight, operative time and remifentanil consumption were comparable among 3 groups. Patients in EAS group had the lowest VAS scores postoperatively among the three groups (P<0.05). The total dose of sufentanil was 115±6.0 μg in EAS group, significantly lower than that in control (134.3±5.9 μg) and sham (133.5±7.0 μg) groups. Similarly, the rescue dose of dezocine was the least in EAS group (P<0.05) among the three groups. Plasma β-EP levels in EAS group at T3 (176.90±45.73) and T4 (162.96±35.00 pg/mL) were significantly higher than those in control (132.33±36.75 and 128.79±41.24 pg/mL) and sham (136.56±45.80 and 129.85±36.14 pg/mL) groups, P<0.05 for all. EAS could decrease the release of PGE2. Plasma PGE2 levels in EAS group at T2 and T3 (41±5 and 40±5 pg/mL respectively) were significantly lower than those in control (64±5 and 62±7 pg/mL) and sham (66±6 and 62±6 pg/mL) groups. Plasma 5-HT levels in EAS group at T2 (133.66±40.85) and T3 (154.66±52.49 ng/mL) were significantly lower than those in control (168.33±56.94 and 225.28±82.03) and sham (164.54±47.53 and 217.74±76.45 ng/mL) groups. For intra-group comparison, plasma 5-HT and PGE2 levels in control and sham groups at T2 and T3, and β-EP in EAS group at T3 and T4 were significantly higher than those at T1 (P<0.05); PGE2 and 5-HT levels in EAS group showed no significant difference among the different time points (P>0.05). No apnea or severe hypotension was observed in any group. It was concluded that intraoperative ipsilateral EAS at PC4 and PC6 provides effective postoperative analgesia for patients undergoing radical esophagectomy with remifentanil anesthesia and significantly decrease requirement for parental narcotics. The underlying mechanism may be related to stimulation of the release of endogenous β-EP and inhibition of inflammatory mediators (5-HT and PGE2).
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adverse effects
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etiology
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Prospective Studies
10.Cure effects of Jiangu Fufang on osteoporotic model induced by ovariectomy.
Ying-Xian DENG ; Zeng-Chun MA ; Hong-Ling TAN ; Cheng-Rong XIAO ; Yu-Guang WANG ; Yue GAO
China Journal of Chinese Materia Medica 2006;31(24):2070-2073
OBJECTIVETo explore the cure effects of Jiangu Fufang on osteoporotic model induced by ovariectomy.
METHODRats were ovariectomized and administered drugs for 3 monthes. Bone mineral density and biomechanics properties, histomorphometric analysis and biochemical index such as calcium, phosphorus, alkaline phosphatase were detected.
RESULTJiangu Fufang could significantly increase bone density and biomechanics properties. The level of calcium, phosphorus and alkaline phosphatase were restored by Jiangu Fufang. Jiangu Fufang could significantly increase area of bone trabecula, thickness of cortical bone and bone trabecula.
CONCLUSIONJiangu Fufang could cure osteoporosis through increasing bone mineral density, improving bone biomechanics properties, and effecting bone metabolism.
Alkaline Phosphatase ; blood ; Animals ; Biomechanical Phenomena ; Bone Density ; drug effects ; Calcium ; blood ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; therapeutic use ; Female ; Femur ; physiopathology ; Lumbar Vertebrae ; drug effects ; pathology ; physiopathology ; Osteoporosis ; blood ; drug therapy ; physiopathology ; Ovariectomy ; Phosphorus ; blood ; Phytotherapy ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley