To develop a method for the determination of residual solvents inα-ketophenylalanine calcium by capillary gas chromatography. Methods:The residual solvents were separated on a DB-624(30 m × 0. 32 mm, 0. 25 μm) capillary chromato-graphic column with temperature programming. The column temperature was maintained at 40℃ for 1 min,and then raised to 180℃at a rate of 10℃·min-1 and maintained for 2 min. N2 was used as the carrier gas, and FID was used as the detector with temperature of 250 ℃. The injector temperature was 200 ℃ and the split ratio was 10∶1, and direct injection was adopted. Methanol, ethanol, ethyl acetate and tetrahydrofuran in α-ketoleucine calcium were detected using an external standard method. Results:The four solvents were separated completely. There was a good linear relationship between the peak area and the concentration of each solvent ( r=0. 997 2-0. 999 5). The average recovery of the four solvents was 95. 47%-100. 26%(RSD≤4. 7%, n=9). Conclusion:The method is rap-id, simple, accurate and sensitive, and can be used in the determination of residual solvents in α-ketophenylalanine calcium.

Objective To develop and validate a simple and reliable HPLC method for the analysis of flee and total carnitine in human serum and to investigate its clinical significance. Methods After proteins in serum were precipitated with a precipitating reagent, carnitine in serum was derivatized to form its ester. HPLC separation of the sample solution was performed on a Lichrospher SiO2 column and detected by ultraviolet absorbance at 260 nm. A mobile phase composed of acetonitrile-citric acid-triethanolamine was found to be the most suitable for this separation at a flow rate of 1.2 ml/min and enabled the baseline separation of the carnitine from interferences with isocratic elution. The free and total carnitine levels in serum were studied in 347 subjects. Results Under the chromatographic conditions described, the carnitine derivative had a retention time of approximately 10 min. Good separation and detectability of carnitine in human serum sample were obtained. The method proved to be linear in the range of carnitine from 0 μmol/L to 400 μmol/L The relative standard deviations of within-assay ( n = 5 ) for free and total carnitine analysis were 3.36% and 1.97% , respectively, and between-assay (n =7) for free and total carnitine analysis were 3.04% and 1.77%, respectively. The average recovery was 98. 2% for free camitine and 96.3% for total carnitine, respectively. The average L-carnitine concentrations in the 347 subjects were as follows: total carnitine (52. 2 ± 8. 6 ) μmol/L, free carnitine ( 42. 3 ± 8. 3 ) μmol/L and acylcarnitine ( 9. 9 ± 2. 9 )μmol/L in the male group ( n = 182), and total carnitine (48.2 ± 9. 9 ) μmol/L, free camitine ( 37.9 ±8. 7) μmol/L and acylcarnitine ( 10. 3 ± 3.5 ) μmol/L in the female group ( n = 165 ). Statistical analysis showed that total and free carnitine levels of male were higher than that of female ( P <0. 01 ) while there was not statistical difference of acylcarnitine levels between two groups ( P > 0. 05). Conclusion The method has been successfully applied to the simultaneous determination of free and total carnitine in serum with good sensitivity, specificity and repeatability, and this is a useful guidance for the clinic therapy and the mechanism study on the diseases associated with carnitine.

Objective To analyze the succumbed reasons of emergent pereutaneous coronary intervention(EPCI)in treatment of acute myocardial infarction(AMI)during operation and in-hospital period.Method During March 1999 to June 2005,623 AMI patients received EPCI,and 27 patients died.The succumbed reasons and clinical characteristics of the succumbed patients were analyzed.Result Among the 27 succumbed patients,with age 51 to 91(69?18)years old, 16 had three-vessel lesions.10 had two-vessel lesions and l had single vessel lesion.Ten patients were accompanied with old myocardial infarction,9 with diabetes meUitus,19 with hypertensions,4 with impaired renal functions,and 6 with old cerebral infarction.Nine patients died of eardingenic shocks,6 died of no-reflows,2 died of heart ruptures,2 thrombosis, 2 acute left heart failure,2 acute renal failure,2 intracranial hemorrhage,l shock due to hemorrhage of puncture position, and l acute perieardiae tamponade.Conclusion The succumbed reasons of EPCI in treatment of acute myocardial infarction were various.Cardiac shock and no-reflow were primary reasons.Old age,multi-vessel lesion,diabetes mellitus, and old myocardial infarction may serve as predicting factors.

Objective:To determine risk factors for postoperative esophageal refractory stenosis after endoscopic submucosal dissection (ESD) of large-scale early esophageal carcinomas and precancerous lesions.Methods:Two hundred and twelve early esophageal carcinomas or precancerous lesions in 186 patients who underwent ESD larger than 3/4 the total esophageal circumference in Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, between July 2013 and December 2017 were divided into two groups according to session number of endoscopic balloon dilatation (EBD), the refractory stenosis group ( n=69, ≥6 EBD sessions) and non-refractory stenosis group ( n=117, ≤5 EBD sessions). Student′s t-test or Mann-Whitney U test was used for univariate analysis and χ2 test and Fisher exact test were used for comparison of categorical variables. Logistic regression was used for multivariate analysis. Results:Compared with the non-refractory stenosis group, the refractory stenosis group had statistically significant differences in the longitudinal diameter of lesions, the longitudinal diameter of artificial ulcer, lesion location, the circumferential range of lesions and the composition of the muscular layer injury (all P<0.05). After eliminating the factor of the vertical diameter of artificial ulcer (because there was significant correlation between the vertical diameter of artificial ulcer and the longitudinal diameter of lesion in clinical practice), multivariate logistic regression analysis showed that the longitudinal diameter of lesion>5 cm (VS ≤5 cm: P=0.003, OR=3.531, 95% CI:1.547-8.060), the location of lesion in the upper thoracic segment (VS lower thoracic segment: P=0.001, OR=36.720, 95% CI:4.233-318.551), in the cervical segment (VS lower thoracic segment: P=0.003, OR=24.959, 95% CI:2.927-212.795), the whole circumferential lesion (VS ≥3/4 but not the whole circumference: P<0.001, OR=10.082, 95% CI:4.196-24.226) and the presence of muscular layer injury ( P<0.001, OR=7.128, 95% CI:2.748-18.486) were more likely to lead to esophageal refractory stenosis after ESD. Conclusion:The longitudinal lesion diameter of more than 5 cm, the circumferential extent of esophageal ESD, cervical or upper-thoracic esophageal lesions, and muscular layer damage are independent risk factors for postoperative esophageal refractory stenosis after ESD for large-scale esophageal cancer and precancerous lesions.

Objective To optimize the culture method for rheumatoid arthritis fibroblast-like synoviocytes in vitro,and observe the effect of Dishevelled (Dvl) 2 on vascular endothelial growth factor (VEGF) in rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS).Methods Synovium from RA patients who underwent knee arthroplasties were cut into small piece,and RA-FLS were isolated and cultured in vitro using tissue block method.Dvl 2 lentivirus overexpressing plasmid was constructed and transfected into RAFLS.Q-polymerase chain reaction (PCR) and enzyme linked immunosorbent assay (ELISA) were used to detect the mRNA and protein expression levels of VEGF.Then we used 10 ng/ml tumor necrosis factor (TNF)-α recombinant protein to stimulate the transfected RA-FLS.24 h after stimulation,mRNA and protein expression of VEGF were detected again.Student's t test was used for two group analyses.Results RA-FLS was successfully isolated and cultured in vitro.The multiplicity of infection was 30 and was in conjunction with appropriate concentration of polybrene to promote transfection.Transfection efficiency could meet the test requirements.The mRNA of Dvl 2 increased for 79-fold than the control group.Compared with the control group,Dvl 2 could mildly inhibit RA-FLS secretion of VEGF.After TNF-α stimulation,Dvl 2 could significantly inhibit the VEGF's mRNA (2.15±0.10,2.92±0.47 fold,t=-3.924,P=0.003) and protein [(285±100) pg/ml,(155±61) pg/ml,t=-2.714,P=0.022] expression compared with the control group.Conclusion Dvl 2 can inhibit the effect of TNF-α induced secretion of VEGF in RA-FLS.The specific mechanism needs further study.

Objective To test the protective immunity in mice induced by recombinant Schistosoma japonicum Sj14FABP through several adjuvant formulations. Methods The recombinant Schistosoma japonicum Sj14FABP was prepared by expression in E. coli as a GST fusion protein (rSj14/GST) and used to vaccinate outbred Kunming mice by using complete Freund's adjuvant (FCA)/incomplete Freund's adjuvant (FIA), Bacillus Calmette-Guerin (BCG) and the immunostimulating complex (ISCOM) as adjuvant respectively. Results The purified recombinant protein rSj14/GST was immunogenic in mice, and 34.3% and 36.0% worm reduction rates were obtained in outbred Kunming mice immunized intradermally with BCG adjuvant and immunized subcutaneously with ISCOM adjuvant respectively, compared with non-vaccinated control group. However, intramuscularly vaccination with rSj14/GST in FCA/FIA was not protective, although the high level of IgG antibody was induced. Conclusion Both BCG and ISCOM are suitable adjuvants for rSj14/GST.

Objective: Oxidative modification of low density lipoprotein(LDL) plays an important role in the development of premature atherosclerosis in autoimmune disorders.Oxidized LDL(ox-LDL) reportedly forms stable complexes with ?2-glycoprotein I(?2-GPI),a major autoantigen for anticardiolipin antibodies,in circulation and the intima of atherosclerotic lesions. This study aims to investigate the serum ?2-GPI/ox-LDL concentration in Chinese patients with systemic lupus erythematosus(SLE),and clinical diagnostic value.Methods: The concentrations of ?2-GPI/ox-LDL complexes were analyzed in 47 SLE patients and 42 healthy controls by ELISA.Results: The serum ?2-GPI/ox-LDL concentrations were significantly higher in the SLE patients than in the controls(\U/ml vs\U/ml,P

Objective To investigate the effects of the interaction between human hepatoma cells and hepatic stellate cells on their growth state,and study its role of interaction on the progression of hepatocellular carcinoma.Methods Human hepatoma cell line HepG2 and hepatic stellate cell line hepatic stallate cells (HSC)-T6 were used and the methods including methyl thiazolyl tetrazolium (MTT) assay,flow cytometry (FCM) analysis,immunohistochemistry,and electron microscopy were employed in this experiment.The effects of conditioned medium (CM) of HepG2 on the activation and proliferation of HSC were explored.The effects of activated HSC CM on HepG2 proliferation were investigated.The uhrastructural changes of the two co-cultured cells were observed.Results MTT assay result showed that HepG2/HSC CM could promote HSC/HepG2 proliferation.FCM result demonstrated that HepG2/HSC CM could influence the cell cycle distribution in HSC/HepG2.Immunohistochemistry exhibited that after the treatment of HepG2/HSC CM,the expression ofα-smooth muscle actin (α-SMA) in HSC and proliferating cell nuclear antigen (PCNA) in HepG2 were increased.When HepG2 and HSC were co-cultured,the ultrastructure of HSC displayed an activated feature.Conclusions HepG2 cells can induce the activation and proliferation of HSC,and the activated HSC can also stimulate the proliferation of HepG2.Interaction between hepatoma cells and hepatic stellate cells may play an important role in the progression of hepatocellular carcinoma.

Objective To retrospectively evaluate the value of SPECT/CT imaging for follow-up of bone metastases.Methods A total of 178 patients with bone metastases (387 lesions) underwent 2 or more events of whole-body bone scintigraphy (WBS) and SPECT/CT imaging.Sequential images were analyzed by 2 experienced,nuclear medicine physicians.Each lesion was interpreted as progressive,remissive or stable in WBS or SPECT/CT independently.Reasons for the discordance between WBS and SPECT/CT results were analyzed.The results of clinical follow-up,including clinical symptoms,tumor markers,serum ALP,radiograph,CT and MRI,were likewise classified as progressive,remissive or stable.The x2 test was used to compare the differences between the two imaging methods.Results The follow-up results of the two imaging methods were consistent in 313 (80.88％,313/387) lesions,including 208 in progression and 105 in remission or stable condition.Among the 74(19.12％,74/387) lesions showing discordance,48 showed remission or stable conditions on WBS but progression on SPECT/CT (64.86％,48/74) ; while 26 showed progression on WBS but remission or stable condition on SPECT/CT (35.14％,26/74).There was a statistically significant difference of the follow-up results between WBS and SPECT/CT (x2 =6.54,P ＜0.05).Conclusion SPECT/CT is more valuable than WBS for follow-up of bone metastases.

To investigate the cellular immunity abnormalities of patients with rheumatoid arthritis(RA), peripheral blood lymphocytes subpopulations, interleukin 2 (IL-2) production and natural killer (NK) cell activity were determined in 9 patients with RA. The results showed that there were a remarkable decrease in NK cell activity and poor response to IL-2 stimlulation. IL-2 production and.the expression of membrane-bound IL-2 receptor were increased in RA patients. The percentage of T4 positive cells and the ratio of T4/T8 were also increased in patients with RA. The results indicate that there are severe cellular immunity abnormalities in patients with RA.