1.Content determination of trace kryptofix 222 in ~(18)F-fluorodeoxyglucose
Journal of Chongqing Medical University 2007;0(08):-
Objective:To establish an ultraviolet spectrophotometry method for determination of trace kryptofix 2.2.2 in 18F-fluorodeoxyglucose product.Method:A storage solution was prepared by adding properly benzaldehyde and lead nitrate to citric acid-NaOH buffer solution(pH = 6.4),and ultralviolet absorbance curve of the solution was changed to platform at the wavelength of 250 nm nearby,and the absorbance value of this solution was 0.4Abs ~ 0.6Abs at 250 nm.Properly 18F-fluorodeoxyglucose product was added to storage solution,and the absorbance value of trace Pb(Ⅱ)-kryptofix 222 complex was detected by ultraviolet Spectrometry for quantitative determination of trace kryptofix 222.Results:The absorbance coefficient of complex was 1.6?104 L/(mol?cm) at 250 nm;the linear range of kryptofix 222 was 0.0?g/ml~10.0?g/ml,the limit ofkryptofix 222 was 0.0?g/ml~10.0?g/ml,the limit of quantification was 0.13?g/ml;the adding recovery was 97.2%~102.7%,with the average recovery of 99.7%;the relative standard deviation(RSD) was 2.4%(n=5).Conclusion:The method was specific,sensitive and suitable for determination of trace kryptofix 2.2.2 in 18F-fluorodeoxyglucose product.
2.Expression, purification and characterization of catalytic domain of protein kinase D1 in baculovirus-insect cell expression system.
Chinese Journal of Biotechnology 2014;30(8):1291-1298
Protein kinase D (PKD) is a novel family of serine/threonine kinases and diacylglycerol (DAG) receptors and has been documented in a variety of cellular processes. To get high purity catalytic domain of PKD1 (PKD1-cat) for crystallography study, the GST-tagged PKD1-cat gene was cloned into a baculovirus transfer vector pFastBac1 (donor plasmid). When the recombinant plasmid was transformed into DH10Bac competent Escherichia coli, which contains a baculovirus shuttle vector (bacmid), transposition occurs to generate a recombinant bacmid containing the gene of interest (GST-PKD1-cat). The recombinant bacmid DNA was transfected into Spodoptera frugiperda Sf9 insect cells to generate recombinant baculovirus, which was then amplified through multiple rounds of infection in Sf9 cells. After that, Trichoplusia ni insect cells in suspension culture were infected with baculoviral stock at a multiplicity of infection (MOI) of 5 PFU/cell. SDS-PAGE and Western blotting analysis confirmed the detection of a 68 kDa protein by the glutathione S-transferase (GST) monoclonal antibody. The recombination protein was purified by Glutathione sepharose affinity chromatography and cleaved by PreScission Protease to remove GST tag, and a highly pure 42 kDa protein which was consistent with the molecular weight of the expected PKD1-cat protein was detected on SDS-PAGE. The activity of purified PKD1-cat protein was determined by in vitro PKD kinase assay. Our data showed that the kinase activity increased with the concentration of purified PKD1-cat protein. These results showed that the truncated recombinant PKD1-cat protein was highly active and pure, and could potentially be used for solving 3D structure of this protein by Nuclear Magnetic Resonance (NMR) or crystallography.
Animals
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Baculoviridae
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Blotting, Western
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Catalytic Domain
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Cell Line
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Electrophoresis, Polyacrylamide Gel
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Genetic Vectors
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Plasmids
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Protein Kinase C
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chemistry
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Recombinant Proteins
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chemistry
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Spodoptera
3.Effects of electroacupuncture at Tianshu (ST25) on pro- and anti-inflammatory cytokines in rats with severe acute pancreatitis.
Qiming XUE ; Lu HUANG ; Ning LI
Journal of Integrative Medicine 2011;9(6):658-664
Objective: To observe the effects of electroacupuncture (EA) at Tianshu (ST25) on pro- and anti-inflammatory cytokines in sodium taurocholate-induced severe acute pancreatitis (SAP) in rats. Methods: Sixty-six male Sprague-Dawley rats were randomly divided into three groups: sham-operation group, SAP group and EA group. An SAP model was established by injecting 3.5% sodium taurocholate at a dose of 1 mL/kg through puncturing the pancreatic duct. In the EA group, EA at ST25 was performed with sparse-dense wave (2/100 Hz, 2 mA for 30 min) immediately after sodium taurocholate injection and 30 min before sacrifice. Rats were sacrificed at 3 (n=7), 6 (n=7) and 12 h (n=8) after operation. Levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-10 in serum were estimated by enzyme-linked immunosorbent assay. Pathological changes of pancreatic tissues were examined by hematoxylin and eosin staining and observed under a microscope and also scored. Results: The levels of serum TNF-α and IL-6 of SAP rats were significantly higher than those of sham-operation rats at 3, 6 and 12 h. The level of IL-10 in serum of SAP rats was significantly higher than that of sham-operation rats at 3 and 6 h (P<0.05). EA at ST25 down-regulated the levels of TNF-α and IL-6 in serum and up-regulated the level of IL-10 and the ratio of IL-10/TNF-α, also attenuated the morphological damages at 6 and 12 h (P<0.05) in rats with SAP. The pathological scores of pancreatic tissues in the EA group were significantly less than those in the SAP group at 6 and 12 h (P<0.05). Conclusion: EA at ST25 may have a therapeutic effect on rats with SAP by down-regulating the levels of TNF-α and IL-6 and up-regulating the level of IL-10 to re-establish the balance of pro- and anti-inflammatory cytokines, and to ameliorate the inflammatory response of SAP.
4.Investigation on Clinical Effect of Weinaian Capsules on Chronic Erosive Gastritis with HP Positivity
Guohong LI ; Xiaoying QIU ; Qiming CHEN ;
Chinese Traditional Patent Medicine 1992;0(07):-
Objective:To investigate the clinical effect of Weinaian Capsules on chronic erosive gastritis with HP positivity and its effect on HP.Methods: 90 patients with chronic erosive gastritis with HP positivity were divided into 2 groups. 50 cases in the treatment group were given drug Weinaian Capsules orally, 4 capsules each time, 3 times a day. 40 cases in the control group were given drug Compound Aluminum Hydroxide Tablets orally, 2 tablets each time, 3 times a day. Both groups were treated for 6 successive weeks. Results: The effective rates were 92% for the treatment group and 67.5% for the control group ( x 2=23.87,P
5.Aquaporin 4 and cerebral edema
Qiming LI ; Dujuan SHA ; Jun ZHANG
International Journal of Cerebrovascular Diseases 2010;18(8):624-631
The aquaporins (AQPs) are a group of highly selective water channel protein family. The content of AQP4 in the astrocytes of the brain is the highest. It is the most vital protein in central nerve system. It participates in a series of pathologic processes of cerebrovascular disorders, brain contusion, tumor, and inflammation caused cerebral edema.However, the mechanism of AQP4 in the process of cerebral edema formation remains controversial. Selective modulation of the expression of AQP4 and intervention in part of its role may provide novel ideas and means for the treatment of edema after cerebral ischemia in clinical practice.
6.Analysis of ~(18)F-fludeoxyglucose Injection by Silica Gel TLC
Qiming LI ; Rongbing JIN ; Xijiang FAN
China Pharmacy 2007;0(25):-
OBJECTIVE: To analyze 18F-fludeoxyglucose injection by Silica Gel TLC method.METHODS: TLC was adopted in which 18F-fludeoxyglucose injection(A) and mixture solution of 18F-FDG injection-acetonitrile(5∶95,V/V)(B) were used as testing solution.GF silica plate and G silica plate were used as stationary phase,and the mobile phase was acetonitrile: water(95∶5,V/V).RESULTS: The main peak Rf of testing solution A had a great fluctuation;The main peak Rf of testing solution B was 0.38~0.40.Radiochemical purity of 18F-fludeoxyglucose was more than 98%,and the Rf of radioimpurity 18F-and the by-product 18F-fludeoxymannose stood at 0.0 and 0.78 respectively.CONCLUSION: When testing solution B,the process of silica gel thin-layer chromatographic analysis is stable and reproducible,and the results meets the requirements specified in USP and are independent of the category of silica plate.
7.Clinical Application and Effective Mechanism of Pacemaker with Atrial Fibrillation Preventive Pacing Algorithms
Shushan QI ; Fangshong LI ; Qiming LIU
Journal of Chinese Physician 2001;0(10):-
Objective To investigate mechanism and clinical effect of implantation of pacemaker with artial fibrillation(Af) preventive pacing algorithms (Vitatron 900E).Method 24 cases of patients with paroxysmal Af were observed in three phases (each phase two weeks):⑴Monitoring phase,in which onset mechanism of atrial arrtythmias was recorded, the device was programmed into DDD mode with 60 bpm;⑵Lower rate limit phase (LRL),DDDR mode with 85bpm LRL, and evaluating anti-Af effect by elevated rate;⑶Preventive pacing therapy phase,all preventive pacing therapies with programming to on or off, and this phase determined whether combined pacing therapies impacted on incidence of Af. Results 24 cases of patients were free of Af, in which 17 cases were done by using post-PAC-response or/and post-exercise-rate-control therapy only, and 7 cases of 17 casees were done by combining it with cordarone 0 1~0 2g/d also. The other 7 cases were done by DDDR 85 bpm only.Conclusions Af prevention pacing may be benefit for patients with paroxysmal Af,especially with complicated symptomatic bradycardia.
8.The correlation between the activity of GSP-? and intestinal metaplasia of gastric mucosa
Yong LIU ; Qiming LI ; Mingzhi LU
Chinese Journal of General Surgery 2001;0(07):-
Objective To study the relationship between the activity of GSP ?, intestinal metaplasia(IM) of gastric mucosa, and proliferation typing.Method Immunohistochemical method was used to detect GSP ? in 167 cases undergoing biopsy or surgery ,among them slices of 112 pathology proved chronic atrophic gastritis(CAG) were counter stained by mucohistochemistry.Results[WT5”BZ] The result showed that the positive rates of GSP ? in gastric adenocarcinoma, CAG with IM or gastric dysplasia were significantly higher than in chronic superficial gastritis (CSG).The positive rate of GSP ? in high proliferation typing (HPT) was significantly higher than in middle or low proliferation typing ( MPT or LPT).Conclusion GSP ? activity could be used as a marker of precancerous lesions and HPT highly correlates to the occurence of gastric cancer.
9.Clinical application of stereotactic radiotherapy in treatment of lung cancer
Dezhi LI ; Ping PU ; Shangzhi FU ; Qiming LI ; Yuping ZHANG
Journal of Third Military Medical University 2001;23(4):473-475
Objective To study the indication, contraindication and short-term clinical effects of stereotactic radiotherapy (X-knife) in lung cancer. Methods A total of 56 cases (Treatment group) of lung cancer were treated with X-knife composing of the modern 3-D treatment planning system and the stereotactic body frame, and SL-75 accelerator. And another 53 cases only treated with routine external radiation served as the control. In treatment group, 42 cases received 50 Gy radiation, at 2 Gy per day, 5 d per week in the upper mediastinum, hilus of lung, below the primary focus first and then underwent X-knife for the primary focuses, and other 14 recurrent and metastatic cases received irradiation of 8-12 Gy per day, 5 d by X-knife alone. Results The actually reexamination rate for 1, 3, 6, and 12 months after X-knife treatment in 78 focuses from 56 cases was 79.5% (62/78), 89.8% (70/78), 87.2% (68/78) and 78.2% (61/78) respectively. The focuses reappeared in 3 months after the treatment and the growth rates of focus in 3, 6, and 12 month after the treatment were 2.6% (2/78), 5.1% and 9% (7/78) respectively. There were 2 focuses received a second treatment with X-knife because no change after the first one. Both the 1-and 2-year survival rates of 36 cases of the primary lung cancer in the treatment group were higher than that of control group (89.1%, 53.9% and 77.4%, 39.6%). Conclusion The stereotactic radiotherapy is of applicable and prospective in the treatment of lung cancer. The indications include: ①As a boost dose in case when primary focus is less than 5 cm after the first external radiotherapy may reduce the exposure of lung tissue to x-ray and the occurrence of radiation pneumonia; ②Treatment for the recurrence after radiotherapy and operation; ③Radical treatment for the metastatic focus less than 5 cm. Attention must be paid to when multiple treatment is carried on the volume of the focus, the Karnofsky scores and the general condition when stereotactic radiotherapy combined with X-knife for lung cancer.
10.Relationship between the plasma brain natriuretic peptide and D-dimer levels and the subtypes of acute cerebral infarction
Qiming LI ; Dujuan SHA ; Jin LI ; Jun ZHANG
International Journal of Cerebrovascular Diseases 2010;18(8):590-594
Objective To investigate the significance of the changes of brain natriuretic peptide (BNP) and D-dimer (DD) levels in patients with acute cerebral infarction with different etiological subtypes. Methods One hundred forty-six patients with acute cerebral infarction within 24 hours of onset were divided into four groups according to the TOAST classification:large-artery atherosclerosis (LAA, n =48), small-artery occlusion (SAO, n = 32), cardioembolism (CE, n = 41 ), and cryptogenic cerebral infarction (n = 25 ). Plasma BNP and DD levels were measured in the emergency department, and the correlation between both plasma BNP and DD levels and different subtypes of cerebral infarction, infarct rolume and severity of disease were analyzed. Results The plasma BNP and DD levels in the cardioembolism group were significantly higher than those in all the non-cardioembolism groups (all P <0.01). The plasma DD level was also increased significantly (compared to the LAA group, P <0. 05; compared to the SAO and cryptogenic cerebral infarction groups, all P < 0. 01 ), while there were no significant differences in plasma BNP and DD levels among all the non-CE groups. The plasma BNP and DD levels in patients of the large infarction group were significantly higher than those of the moderate infarction group (t = 3.766 and 3.029, respectively; P = 0. 013 and 0. 029,respectively), and there was no significant difference between the moderate infarction group and the small infarction group (t= 1.275 and 1.207, respectively; P= 0.258 and 0. 281,respectively). The plasma BNP and DD levels in the National Institutes of Health Stroke Scale (NIHSS) score ≥ 7 group were significantly higher than those in the NIHSS score < 7 group (t = - 3.454 and - 4. 044, respectively; P = 0. 018 and 0. 010, respectively). Conclusions Acute cerebral infarction, particularly the plasma BNP and DD levels were increased significartly in patients with cardioembolism, and the larger the infarct volume, the more serious the disease, and the higher the both levels. Early detection of plasma BNP and DD levels contribute to etiological classification of cerebral infarction, especially for the diagnosis of cardioembolism, as well as the identification of infarct volume and the severity of the disease.