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.

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 ; Baculoviridae ; Blotting, Western ; Catalytic Domain ; Cell Line ; Electrophoresis, Polyacrylamide Gel ; Genetic Vectors ; Plasmids ; Protein Kinase C ; chemistry ; Recombinant Proteins ; chemistry ; Spodoptera

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.

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.

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.

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.

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.

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

Objective Objective To investigate the clinical application of high-dose-rate(HDR)endobronchial brachytherapy for the treatment of primary lung cancer with atelectasis. Methods A total of 142 cases of lung cancer with atelectasis treated with HDR endobronchial brachytherapy and 71 cases(contr01)treated with extemal irradiation only at onr center of cancer were comparatively studied.1n the treatment group,with the aid of bmnchoscope,brachytherapy was applied 2 or 3 times at the dose of 8 Gy each time to a tissue depth of 1 to 1.5 cm.Cases in the contmlgroup,treated with external irradiation,received 60-70 Gy in total.Results Complete recruitment of atelectasis was observed in 80.28%cases of the treatment group and 64.79%in the control,T11ere Was significant difference between the two groups.Survival rates after 1,2 and 3 years in cases with complete remission of atelectasis in treatment group were 60.68%,44.44%and 23.93%,but 36.00%,28.00%and 16.00%in cases with no remission of atelectasis.Significant difference of one-year survival Was found between the two groups(P

Objective:To optimize the ultrasonic extraction process of Danggui Baogan capsules. Methods:The ultrasonic extrac-tion process of Danggui Baogan capsules was optimized by L9 (34 ) orthogonal test with the contents of ferulic acid and angelica sinensis polysaccharide as the indices and the amount of 30% ethanol, ultrasonic duration and extraction times as the influencing factors. Re-sults:The best ultrasonic extraction conditions were as follows: adding 12-fold amount of 30% ethanol and extracted four times with 1. 5 hours for each time. Conclusion:The ultrasonic extraction process is simple and reproducible, and suitable for the industrial pro-duction.