Objective: To establish a capillary electrophoresis (CE) with electrophoretically mediated microanalysis (EMMA) method for the determination of EDTA-2Na in Japanese encephalitis attenuated live vaccine.
Methods: The test was performed in disodium hydrogen phosphate buffer with pH 2.5, the online metal ions complexation of 1.5 mg·mL^-1 Fe³⁺ and incubation time of 3 min. The separation voltage was 25 kV, the detection wavelength was 257 nm, and. the column temperature was 25.0 ℃.
Results: The established method had a good linear relationship in the concentration range of 0.01-0.5 mg·mL^-1 (r=0.999 9), the detection limit was 5 μg·mL^-1, and the relative standard deviation (RSD) of the measured samples was less than 2.87%. The recoveries of spiked samples were between 96.49%-101.02%.
Conclusion:The optimized method was applied to the determination of EDTA-2Na in Japanese encephalitis attenuated live vaccine. The satisfactory experimental results were obtained.

Objective:To explore the mechanism of early pancreatic exocrine function changes in severely scalded rats.Methods:The experimental research methods was used. Eighty male Sprague-Dawley rats aged 7-8 weeks were divided into simple sham injury group ( n=8), sham injury+cholecystokinin octapeptide (CCK8) group ( n=8), severe scald+CCK8 group ( n=32), and extremely severe scald+CCK8 group ( n=32) by the random number table, which were treated accordingly. Immediately after injury of rats in the 2 sham injury groups and 1, 2, 3, and 7 days after injury of rats in the 2 scald groups, the improved methods including pancreatic duct puncture and catheterization were used to dynamically collect the pancreatic-bile juice (PBJ) of rats. The PBJ secretory volume within 1 h was recorded, and the content of pancreatic lipase, α-amylase, and trypsin in PBJ was detected by enzyme-linked immunosorbent assay (ELISA), and the number of samples was 8. The femoral venous blood was collected, and the concentrations of pancreatic lipase and α-amylase in serum were detected by standard colorimetry to reflect their activity ( n=8). The pancreatic tissue was extracted, and the levels of interleukin-1β (IL-1β) and IL-6 in pancreatic tissue were detected by ELISA ( n=8), the expression of hypoxia-inducible factor 1α (HIF-1α) in pancreatic tissue was detected by immunofluorescence method, and the histopathological changes in pancreatic tissue were observed by hematoxylin-eosin staining, the severity of pancreatic tissue injury in the 2 scald groups was evaluated by modified Schmidt method ( n=6), and the ultrastructure of acinar cells in pancreatic tissue was observed by transmission electron microscopy. Data were statistically analyzed with analysis of variance for factorial design, Tukey test, independent sample t test, and least significant difference test. Results:Compared with the PBJ secretory volume (0.740±0.030) mL in the pancreatic tissue of rats in simple sham injury group within 1 h immediately after injury, the (0.823±0.033) mL in sham injury+CCK8 group was significantly increased ( t=4.92, P<0.05). Compared with that of rats in sham injury+CCK8 group immediately after injury, the PBJ secretory volume of rats within 1 h in severe scald+CCK8 group ((0.681±0.024), (0.608±0.056), (0.525±0.025), and (0.720±0.044) mL) and extremely severe scald+CCK8 group ((0.540±0.025), (0.406±0.021), (0.475±0.036), and (0.690±0.018) mL) was significantly decreased on 1, 2, 3, and 7 days after injury ( P<0.05). Compared with that in severe scald+CCK8 group, the PBJ secretory volume of rats within 1 h in extremely severe scald+CCK8 group was significantly decreased on 1 and 2 days after injury ( P<0.05). Compared with that of rats in simple sham injury group immediately after injury, the content of pancreatic lipase, α-amylase, and trypsin in PBJ of rats in sham injury+CCK8 group immediately after injury was significantly increased (with t values of 4.56, 3.30, and 4.99, respectively, P<0.05). Compared with that of rats in sham injury+CCK8 group immediately after injury, the content of pancreatic lipase and α-amylase in PBJ of rats in severe scald+CCK8 group and extremely severe scald+CCK8 group was significantly decreased on 1, 2, 3, and 7 days after injury ( P<0.05), the trypsin content in PBJ of rats in extremely severe scald+CCK8 group was significantly decreased on 2 days after injury ( P<0.05). Compared with that in severe scald+CCK8 group, the content of pancreatic lipase in PBJ of rats in extremely severe scald+CCK8 group was significantly decreased on 1, 2, and 3 days after injury ( P<0.05), and the content of α-amylase and trypsin in PBJ was significantly decreased on 1 and 2 days after injury ( P<0.05). There were no statistically significant differences in the activities of pancreatic lipase and α-amylase in serum of rats among the 4 groups at various time points after injury ( P>0.05). Compared with that of rats in sham injury+CCK8 group immediately after injury, the levels of IL-1β in pancreatic tissue of rats in severe scald+CCK8 group on 1, 2, and 3 days after injury and in extremely severe scald+CCK8 group on 1, 2, 3, and 7 days after injury were significantly increased ( P<0.05), and the levels of IL-6 in pancreatic tissue of rats in severe scald+CCK8 group and extremely severe scald+CCK8 group were significantly increased on 1, 2, 3, and 7 days after injury ( P<0.05). Compared with that in severe scald+CCK8 group, the IL-1β level in pancreatic tissue of rats in extremely severe scald+CCK8 group was significantly increased on 2 and 3 days after injury ( P<0.05), and IL-6 level in pancreatic tissue was significantly increased on 2 days after injury ( P<0.05). The expression levels of HIF-1α in pancreatic tissue of rats in simple sham injury group and sham injury+CCK8 group immediately after injury were lower; and compared with that in sham injury+CCK8 group immediately after injury, the expression levels of HIF-1α in pancreatic tissue of rats in the 2 scald groups increased to a certain extent at different time points after injury, and the expression position was transited from the edge of the pancreatic tissue to the whole pancreas, the expression levels of HIF-1α in pancreatic tissue of rats in the 2 scald groups tended to be normal on 7 days after injury. Compared with that in simple sham injury group immediately after injury, the proportion of acinar cell cytoplasm in pancreatic tissue of rats in sham injury+CCK8 group was increased; and with the increase of time after injury, edema, hemorrhage, necrosis, and inflammatory infiltration appeared in pancreatic tissue of rats in the 2 scald groups. Compared with that in severe scald+CCK8 group, the scores of edema, inflammatory cell infiltration, bleeding, and necrosis in pancreatic tissue of rats in extremely severe scald+CCK8 group were increased to varying degrees at various time points after injury, and the scores of pancreatic tissue of rats in the 2 scald groups basically recovered to normal on 7 days after injury. Compared with that in simple sham injury group immediately after injury, the number of enzyme granules in acinar cells of pancreatic tissue of rats in sham injury+CCK8 group was increased, and with the increase of time after injury, the enzyme granules in acinar cells of rats in the 2 scald groups were gradually reduced basically. Conclusions:The exocrine functions of pancreas, such as synthesis and secretion of pancreatic enzymes, are decreased in the early stage in severely scalded rats. And the greater the scalded area, the more significant the decline of pancreatic exocrine function. This change may be related to hypoxic injury and inflammation in pancreatic tissue after severe scald.

Objective:To explore the characteristics of intestinal flora in female patients with type 2 diabetes mellitus (T2DM) with comorbid coronary heart disease.Methods:Female patients with T2DM from September 2019 to November 2020 were enrolled in this study and divided into 2 groups stratified by coronary heart disease: 22 patients with coronary heart disease(T2DM+ CHD group) and 49 patients with simple T2DM group(T2DM group). Thirty-one healthy females were selected as the normal control group(NC group). The abundance of intestinal flora, the difference in intestinal flora and its relationship with indicators such as glycosylated hemoglobin and fasting blood glucose was analyzed by the real-time fluorescent quantitative PCR detection technology in the three groups.Results:The abundance of Prevotella in the T2DM group was lower, and Roseburia inulinivorans and Faecalibacterium prausnitzii in the T2DM+ CHD group were lower, while Bacteroides and Enterococcus spp in the T2DM+ CHD group were higher compared with NC group, respectively( P<0.05). The abundance of Bacteroides and Enterococcus spp in the T2DM+ CHD group was lower than that in the T2DM group( P<0.05). Spearman correlation analysis showed that the abundance of Faecalibacterium prausnitzii and Eubacterium rectale was negatively correlated with obesity while abundance of Bacteroides and Enterococcus spp was positively correlated with HbA 1C and fasting blood glucose. The abundance of Bacteroides was positively correlated with TCH and TG(all P<0.05). Logistic regression analysis showed that Bacteroides and Enterococcus spp were independent influencing factor of the development of female patients with T2DM and CHD. Conclusion:Female T2DM and T2DM with CHD had intestinal flora imbalance, which was related to a variety of glucose and lipid metabolism indicators, and might be closely related to the occurrence of T2DM and CHD. Bacteroides and Enterococcus spp were independent influencing factors of the development of female patients with T2DM and CHD. Regulating the intestinal flora can provide ideas for the prevention and treatment of T2DM with CHD in female.

Objective:To prospectively follow up the patients with ileocecal inflammatory lesions, to explore the characteristics of Crohn′s disease(CD) at early stage, and to provide references for early diagnosis of CD.Methods:From January 2013 to December 2018, at Department of Gastroenterology, The Seventh Medical Center of PLA General Hospital, 232 patients with unexplained ileocecal inflammatory lesions under colonoscopy examination were enrolled, which were followed up for more than one year. Chi-square test and Fisher exact probability text were used to compare the patients with early CD, with non-specific enteritis and intestinal tuberculosis in abdominal symptoms (abdominal pain, diarrhea, abdominal distension, constipation, hematochezia, changes in bowel habits), accompanying symptoms (oral ulcer, arthralgia), the proportion of patients with elevated erythrocyte sedimentation rate (ESR) or elevated C-reactive protein (CRP) level, serum antineutrophilic cytoplasmic antibody (ANCA), anti-saccharomyces cerevisiae antibody (ASCA), tuberculosis infection of T cells spot test, positive rate of fecal occult blood, lesion size, morphology, involvement site under endoscopy and histopathological results. Multivariate binary logistic regression was used to analyze the related factors of early CD.Results:Among 232 patients, 155 were males and 77 were females, and the age of first diagnosis was (43.9±13.8) years old. The follow-up period (range) was 27 months (12 to 79 months). Twenty-nine cases (12.5%) were diagnosed as early CD, 45 cases (19.4%) were intestinal tuberculosis, 105 cases (45.3%) were non-specific enteritis, and 53 cases (22.8%) as undetermined. All of 29 patients with early CD had abdominal symptoms, which accounted for 16.9% (29/172) of 172 patients with ileoceccal inflammatory lesion as well as abdominal symptoms. In early CD patients, the proportions of patients with abdominal pain, elevated CRP level and ESR level, positive rate of ASCA, positive rate of tuberculosis infection T cells and percentage of patients with thickened intestinal wall were all higher than those in patients with non-specific enteritis (62.1%, 18/29 vs. 33.3%, 35/105; 13.8%, 4/29 vs. 0; 13.8%, 4/29 vs. 1.0%, 1/105; 24.1%, 7/29 vs. 1.0%, 1/105; 20.7%, 6/29 vs. 3.8%, 4/105; 95.7%, 22/23 vs. 0), and the proportion of patients without abdominal symptoms was lower than that of patients with non-specific enteritis (0 vs. 31.4%, 33/105). And the differences were statistically significant ( χ2=6.692, Fisher exact probability text, χ2=7.162, χ2=17.826, χ2=7.497, Fisher exact probability text, and Fisher exact probability text, all P<0.05). Early CD patients were more likely to have multiple lesion sites (55.2%, 16/29), and mainly deep ulcers (55.2%, 16/29) and ulcers with a long diameter of 5 to 10 mm (39.3%, 11/28). The lesions of non-specific enteritis were mostly confined to the end of ileum (75.2%, 79/105), which were mainly superficial ulcers (41.0%, 43/105) and ulcers with a long diameter less than 5 mm (69.0%, 49/71). The proportion of patients without abdominal symptoms and the positive rate of tuberculosis infection of T cells spot test of early CD patients were both lower than those of intestinal tuberculosis group (0 vs. 15.6%, 7/45 and 20.7%, 6/29 vs. 68.9%, 31/45). The positive rate of ASCA and the proportion of patients with thickened intestinal wall were higher than those of intestinal tuberculosis group (24.1%, 7/29 vs. 0 and 95.7%, 22/23 vs. 11/19), and the differences were statistically significant (Fisher exact probability text, χ2=13.713, Fisher exact probability text and χ2=6.710, all P<0.05). The results of multivariate binary logistic regression analysis showed that abdominal pain and positive ASCA were independent risk factors for early CD (odds ratio ( OR)=2.855, 95% confidence interval ( CI) 1.014 to 8.037, P=0.047; OR=10.033, 95% CI 2.274 to 44.250, P=0.002). Conclusions:Prospective follow-up for more than one year in patients with unexplained ileocecal inflammatory lesions can effectively identify and diagnose early CD. Ileocecal inflammatory lesions with abdominal symptoms are one of the early manifestations of CD. Abdominal pain and positive serum ASCA at the initial diagnosis are independent risk factors for early diagnosis of CD.

Objective To study the differences of endoscopic submucosal dissection ( ESD ) for colorectal tumors of different diameters. Methods Data of 210 cases which were treated with ESD for colorectal tumors at the Endoscopy Center, the Seventh Medical Center of PLA General Hospital from October 2012 to December 2015 were retrospectively analyzed. The lesions were divided into two groups according to different diameters (≥4. 0 cm group and <4. 0 cm group) for comparative analysis of related factors. Results The mean procedure time of ESD for 210 colorectal tumor cases was 50. 3±42. 7 min and the mean size of lesions was 7. 98 ± 10. 84 cm2 . En bloc resection rate was 91. 4％, R0 resection rate was 90. 5％, and the curative resection rate was 88. 6％. Perforation rate was 5. 2％ (11/210), and the late hemorrhage rate was 0. 5％ (1/210). Compared with lesions < 4. 0 cm, those ≥ 4. 0 cm required longer resection time (79. 63±53. 91 min VS 35. 28±24. 99 min, P<0. 001); and the lesions were mainly located in the rectum ( 61. 97％) . LSTs were mainly mixed granular/nongranular type ( 54. 93％);en bloc resection rate, complete resection rate and curative resection rate of the tumors≥4. 0 cm were all lower than those of tumors < 4. 0 cm. The difference in complete resection rate was statistically significant ( 85. 92％ VS 94. 24％;P=0. 041) . The perforation rate ( 7. 04％) was higher in≥4. 0 cm group, but the difference was not statistically significant. Conclusions ESD of colorectal tumors of diameters ≥ 4. 0 cm requires longer time with higher operation risk. Additionally, physicians should be more careful with non-rectal lesions.

Objective To explore the value of methylation of SDC2 and SFRP2 genes promoter in fecal DNA for colorectal cancer ( CRC) screening. Methods All stool samples were enrolled from Changhai Hospital of Naval Medical University, the Tenth People' s Hospital of Tongji University and the Seventh Medical Center of Chinese People's Liberation Army General Hospital. A total of 500 stool samples collected from March 2018 to December 2018 were allocated to CRC group ( 132 CRCs ) , adenoma group ( 38 advanced adenomas), healthy group (152 healthy individuals), interferential group (178 cases of benign colorectal disease or other non-colorectal tumors) and negative group (330 cases composed of healthy group and interferential group ) . The promoter methylation of fecal SDC2 and SFRP2 genes was detected by methylation-specific PCR (MSP) and compared with single gene methylation and the fecal immunochemical tests ( FIT) to evaluate its sensitivity and specificity. Results The stool sample analysis showed that the sensitivity of combined detection of SDC2 and SFRP2 in CRC group was 97. 73％ ( 129/132 ) , which was significantly higher than those of the single gene SDC2 test [ 70. 45％ ( 93/132) , P=0. 000] , single SFRP2 test [81. 82％ (108/132), P=0. 000] and FIT [69. 70％ (92/132), P=0. 000]. In adenoma group, the sensitivity of combined detection of SDC2 and SFRP2 was 57. 89％ (22/38), which was significantly higher than those of the single gene SDC2 test [ 15. 79％ ( 6/38 ) , P= 0. 000 ] and FIT [ 21. 05％ ( 8/38 ) , P=0. 021] , with no significant difference compared with that of SFRP2 test [ 47. 37％ ( 18/38) , P=0. 358] . In healthy group, the specificity of combined detection of SDC2 and SFRP2 was 98. 68％ (150/152), with no significant difference compared with those of single gene SDC2 test [ 100. 00％( 152/152) , P=0. 156] , single SFRP2 test [98. 68％ (150/152), P=1. 000] or FIT [95. 39％ (145/152), P=0. 091]. Specificities of combined detection of two genes in interferential and negative groups were 90. 45％ ( 161/178) and 94. 24％( 311/330) , which were significantly higher than 73. 03％( 130/178, P=0. 000) and 83. 33％( 275/330, P=0. 000) of FIT, respectively. Conclusion The combined detection test of methylation of SDC2 and SFRP2 is superior to single gene test, whose sensitivity of CRC and aggressive adenoma and specificity of distinguishing benign and malignant lesions are higher than FIT, which has potential application value.

ObjectiveTo explore the feasibility and safety of preoperative cardiac rehabilitation (CR) nursing intervention in patients with severe valvular disease, and to evaluate the effect of preoperative exercise-mediated cardiac rehabilitation nursing on the clinical outcomes of patients after operation. MethodsFrom January to December of 2017, 108 patients with advanced valvular heart disease (AVHD) admitted in Department of Cardiothoracic Surgery, Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School were selected based on the diagnostic standard for AVHD. The AVHD patients should not be younger than 18 years old, left ventricular ejection fraction(LVEF) less than 40%, and need repair of interventricular septal perforation, resection of ventricular aneurysm and radiofrequency ablation of surgical atrial fibrillation simultaneously. Random number method was used to divide the patients into control group and cardiac rehabilitation nursing intervention group (CR group). Mitral and aortic valve lesions were the main types of valvular lesions in the two groups, and the principles of preoperative treatment were the same. Patients in the control group received routine nursing after admission. The patients in the observation group were evaluated by 6 minute walking test (6MWT) after admission. According to the distance of 6MWT, MOTO Med nursing intervention was carried out in gradeⅠ andⅡ. Frequency was once a day, 30 minutes each time, the first movement resistance was 1 kg, and was adjusted appropriately according to the individual situation. Exercise training and aerobic training were carried out for gradeⅢ andⅣ, with emphasis on lower limb muscles, abdominal muscles and ectopectoralis once a day for 20 minutes each time. ResultsThere was no significant difference in hospital stay, ICU stay time and mechanical ventilation time between the two groups (P＞0.05). The LVEF value of CR group was significantly higher than that of control group at the early stage after operation [(42.5±6.2) vs .(31.0±4.5),P＜0.001)]. The 6 MWT (P=0.009) and self-management(Barthel) score (P=0.001) of CR group were better than those of control group. There was no significant difference in quality of life between the two groups 30 days after operation(P＞ 0.05). ConclusionsPreoperative cardiac rehabilitation nursing intervention can effectively improve the cardiac function of patients with severe valvular disease early after operation, increase the early exercise tolerance of patients after operation, improve the early self-care ability of patients after operation, and help patients recover better and faster.

OBJECTIVE:To establish a method for the determination of dendrobine and its metabolites M-250 and M-280 in mice plasma for the first time. METHODS:Mice were given dendrobine 60 mg/kg by intragastric administration,1 h later plasma were collected and treated. Using pseudoephedrine hydrochloride as internal standard and dendrobine reference substance as control, the plasma concentrations of dendrobine and its metabolites M-250 and M-280 were determined by UPLC-MS combined with quantitative analysis of multi-components by single marker. The separation was performed on Hypersil Gold C18 column with 0.1％formic acid-acetonitrile(gradient elution)at the flow rate of 0.3 mL/min. The column temperature was set at 40℃,and sample size was 5 μL. Heatable electrospray ionization (HESI) source, scan/ESI + were applied and operated in positive ion mode with atomization temperature of 300℃,ion transmission tube temperature of 350℃,the sheath gas velocity of 35 arb,the auxiliary air velocity of 15 arb,the spray voltage of 3.5 kV,the collision voltage of 30,40,50 eV. The mass-to-charge ratio of detection range were 100-1500. RESULTS:The endogenous substances of mice plasma had no interference with the content determination of dendrobine and its metabolites M-250 and M-280. The linear range of dendrobine were 9.13-912.94 ng/mL(r=0.9996). The limit of quantitation was 3.04 ng/mL. RSDs of intra-day and inter-day were all less than 7.5％(n=5 or n=3). The accuracy were 96.8％-107.5％(n=5). Matrix effects were 97.1％-106.0％(RSD=1.8％-4.7％,n=5). RSDs of the content of sample at 15℃ for 24 h,at -70 ℃ after three times freeze-thaw,at -70 ℃ for 15 d were lower than 12.8％ (n=3). The content of dendrobine in plasma sample of mice was (41.3 ± 5.7) ng/mL (n=12). The contents of its metabolites M-250 and M-280 were (493.0 ± 73.1) and (41.4 ± 3.0) ng/mL (n=12) with Relative correction factor of 1.0. CONCLUSIONS: The method is sensitive and accurate,and can be used for content determination of dendrobine and its metabolites M-250 and M-280 in mice plasma.

Objective To discuss the clinical effects and the major adverse cardiac events of intracoronary prostaglandin E1 injection prior to percutaneous coronary intervention (PCI) in early(within 24h of symptom onset) interventional treatment for patients with acute non -ST segment elevation myocardial infarction ( NSTEMI ) . Methods 122 patients with NSTEMI who underwent early interventional treatment were divided into three groups according to the digital table:41 cases in prostaglandin E 1 group,41 cases in nitroglycerin group ,40 cases in control group.The TIMI blood flow was compared among the three groups after PCI .All patients were followed up during 6 months about major adverse cardiac events ( MACE) and the cardiac structure and function by echocardiography . Results After primary PCI,the corrected TIMI frame count(CTFC) was significantly better in the prostaglandin E 1 group[(20.22 ±6.82)] than in the nitroglycerin group[(26.35 ±8.71)] and the control group[(27.02 ±9.65), t=6.451,6.763,all P<0.05].The TIMI myocardial perfusion grade (TMP) was significantly better in the prosta-glandin E1 group(7.3％) than in the nitroglycerin group(26.8％) and the control group(30.0％)(P<0.05). There was no statistically significant difference between the nitroglycerin group and the control group (P>0.05).All patients were followed up for 6 months,the LVDd in the prostaglandin E1 group[(46.8 ±3.7)mm] was significantly lower than that in the nitroglycerin group[(49.5 ±5.8) mm] and the control group [(50.2 ±4.9) mm,t=6.312, 5.893,all P<0.05].The LVEF in the prostaglandin E1 group [(55.8 ±8.2)％] was significantly higher than that in the nitroglycerin group [(49.3 ±7.9)％] and the control group [(50.5 ±6.8)％,t=7.011,5.981,all P<0.05].The incidence rate of MACE in the prostaglandin E 1 group(4.9％) was significantly lower than that in the nitroglycerin group(12.2％) and control group(12.5％)(χ2 =5.834,5.719,all P<0.05).There was no statistically significant difference between the nitroglycerin group and the control group (P>0.05).Conclusion Intracoronary administration of prostaglandin E 1 injection prior to balloon dilation can significantly improve the myocardial microcir-culation perfusion,and can decrease MACE in patients with NSTEMI who underwent early interventional treatment .

Objective To evaluate the clinical efficacy of pre-procedure simethicone on detection of pharynx by upper gastrointestinal endoscopy. Methods A total of 100 patients with esophageal squamous cell carcinomas ( ESCCs) were enrolled in this prospective controlled trial and randomly assigned into two groups. The study group was given gargle with 5 mL simethicone plus 5 mL water combined with conventional procedure before gastroscopy. The control group was prepared according to the conventional procedure. The age, gender, tumor stage, lesion size, doctor＇s satisfaction and examination time between two groups were analyzed. Results The basic conditions between the two groups, including age, gender, and tumor stage were not significantly different (all P＞0. 05). The detection rate of superficial lesion in pharynx was higher in the study group than that in the control group, with no significant difference [8. 16%(4/49) VS 2. 04%( 1/49), P＝0. 362]. The median time of pharyngeal observation in the study group was less than that in the control group (21. 7 s VS 33. 9 s, P＝0. 000). The doctor was more satisfied in the study group than the control group (P＝0. 001). Conclusion Pre-procedure with simethicone improves the endoscopic visibility and detection rate of superficial squamous cell carcinoma with less observation time.