An online solid phase extraction ( SPE ) coupled with high performance liquid chromatography (HPLC)-mass spectrometry (MS) method was developed for the separation of 1,3,5,7-tetraacetyl-1,3,5,7-tetraazacyclooctane ( TAT ) and 1 , 3 , 5-triacetyl-1 , 3 , 5-triazacyclohexane ( TRAT ) which are the synthetic intermediates of cyclotetramethylenetetranitramine ( HMX) . In this experiment, molecularly imprinted polymers with TAT as the template were used as SPE sorbents. PC HILIC column was employed in liquid chromatographic separation. The parameters of SPE-HPLC were optimized. Acetonitrile was selected as the loading solution with flow rate of 0. 1 mL/min. After flushed by ethyl acetate, the TAT adsorbed on SPE was eluted by methanol, which was also used as the mobile phase in HPLC separation. The mass spectrometry was coupled with HPLC to identify the corresponding peaks. Under the optimized conditions, the linear detection range of this method was 6. 0 mg/L to 500. 0 mg/L, with the detection limit of 1. 8 mg/L (3σ). The enriching factor was 400 times and TAT recovery was 79%–93% in the standard addition experiment.

Professor TANG Wei-yong's treatment for children acute tonsillitisis from the three aspects oftreating symptoms, treating muscle and reconciliation, namely three-solution method. In addition, he modified three ancient prescriptions to create three-solution prescription. He applied the prescription into clinical practice and obtained a lot of good efficacy.

Objective The toxicological compatibility of Aconiti Carmichaeli Radix, as a toxic Chinese medicinal herb, combined with the other ingredients in Sini Decoction was investigated to elucidate the rationality of the combination of the ingredients in Sini Decoction from toxicological point of view. Methods Three kinds of experiments, acute toxicity in mice, heart toxicity in rats and aconitines level in water extract of Sini Decoction and its ingredients including Aconiti Carmichaeli Radix alone and its combination with licorice or dried ginger were adopted in this study. In the toxicological experiments, LD50 values for the acute toxicity test and TD50 values for the heart toxicity (arrhythemia as a parameter) of Sini Decoction, Aconiti Carmichaeli Radix alone and its combination with licorice or dried ginger were comparatively determined. And levels of individual aconitines of the water extracts from Sini Decoction, Aconiti Carmichaeli Radix alone and its combination with licorice or dried ginger were measured, respectively. Results The LD50 and TD50 of the combination of Aconiti Carmichaeli Radix and licorice in Sini Decoction were found to be higher than Aconiti Carmichaeli Radix alone or Sini Decoction, while the LD50 and TD50 of the combination of Aconiti Carmichaeli Radix and dried ginger appeared to be not different from those of Aconiti Carmichaeli Radix alone. The level of the main toxic compound of the water extracts for the combination of Aconiti Carmichaeli Radix and licorice, and Sini Decoction was lower than that of Aconiti Carmichaeli Radix alone and its combination with dried ginger. Conclusion The combination of Aconiti Carmichaeli Radix and licorice can attenuate the toxicity of Aconiti Carmichaeli Radix.

Objective To investigate the safety and efficacy on early treatment of large pancreatic pseudocyst by endoscopic ultrasound (EUS)-guided transgastric drainage. Methods The clinical data of 23 cases of large pancreatic pseudocyst treated with EUS-guided transgastric drainage from 2003 to 2008 was retrospectively analyzed. Results All were of single pseudocyst. Pseudocyst was present in the head of the pancreas in 3 cases, in the body of the pancreas in 11 cases, in the tail of the pancreas in 9 cases. Mean diameter of pancreatic pseudocysts was 11 cm( range 8 - 18 cm ). The interval between the finding of pseudocysts and EUS-guided transgastric drainage ranged 17 -65 d, averaging 31 d. Two had postoperative infection of the pseudocyst, one received percutaneons external drainage and the other one received surgical internal drainage. Three had postoperative gastrointestinal bleeding and were treated with antacid and hemostatic drugs. Patients were examined by CT two to three months later. Pancreatic pseudocysts disappeared completely in 6 cases and significantly shrank in the rest. The clinical symptoms of all patients,such as abdominal pain, abdominal bloating were significantly relieved. All patients were followed-up for 1 year, there was no pseudocyst recurrence, ulcer,bleeding or infection. Conclusions Early treatment of large pancreatic pseudocyst by EUS-guided transgastric drainage is safe and effective.

OBJECTIVE: To observe the clinical effect of 5% D-fructose injectio on energy supply in surgery patients. METHODS: By setting 5% glucose injectio as control,the influence of 5% D- fructose injectio on blood sugar level,liver and kidney function indices was detected.RESULTS: 5% D-fructose injectio did not influence liver and kidney functions, serum uric acid and RESULTS: of routine examination of blood and urine.Compared with control group, the change of blood sugar level in experiment group was slighter.CONCLUSION: 5% D-fructose injectio is effective and safe for energy supply in surgery patients.

Objective: To investigate the general situation of scoliosis and influencing factors among middle school students in Guangzhou, so as to provide evidence for behavioral intervention measures. Methods: By stratified cluster random sampling method, 2 121 students from 8 middle schools of Guangzhou were selected to conduct questionnaire survey and physical examination. Results: The detection rate of scoliosis among middle school students in Guangzhou was 8.20%, girls (9.50%)＞boys (7.00%), urban area (10.45%)＞suburb area (4.77%), senior high school (10.08%)＞junior school (6.39%) ( P ＜0.05). Among the most of the 174 positive students were found to have large thoracic curve, right lateral bending, being girls with medium scoliosis. Multivariate Logistic regression analysis showed that students in urban area ( OR=2.56, 95%CI =1.71-3.82), with mobile electronic devices usage time≥3 h/d( OR=1.59, 95%CI =1.12-2.27), prolonged near vision work ≥1 h ( OR=1.40, 95%CI =1.00-1.95), outdoor activity time＜2 h/d( OR=1.82, 95%CI =1.24-2.67) had a higher detection rate of scoliosis ( P <0.05). Conclusion The detection rate of scoliosis among middle school students in Guangzhou is much higher, which might be related to district, mobile electronic devices usage, prolonged near vision work and insufficient outdoor activity. Health education regarding spinal knowledge should be strengthened.

Cloning of flanking sequences of double-copy gene is a challenge in molecular biology. We developed a method to solve this problem by combining an optimized inverse PCR (iPCR) with TAIL-PCR. First, Southern blotting analysis was used to determine a proper restriction enzyme that could obtain proper-length restriction fragments that contained the target gene. Then optimized iPCR was performed to amplify the restriction fragments that contained the separated copies of the gene. Based on the obtained sequences, TAIL-PCR was performed to amplify further flanking regions of the gene. With this method, we obtained all of the EcoR I restriction fragments (2.2-5.1 kb) and Hind III restriction fragments (8.5-11.7 kb) of mitochondrial atpA gene in cytoplasmic male sterile (CMS) line and maintainer line of Upland cotton. The results showed that this method was an efficient approach to clone flanking sequences of double-copy gene.
Chromosome Walking ; Cloning, Molecular ; Gene Expression Regulation, Plant ; Genes, Mitochondrial ; Genes, Plant ; genetics ; Gossypium ; genetics ; Plant Proteins ; genetics ; metabolism ; Polymerase Chain Reaction ; methods ; Terminal Repeat Sequences

Objective: To study on the mRNA expression level of IN1 gene associated with the clinical efficacy and prognosis of hepatoblastoma(HB) in children and to elucidate the early warning value of IN1 gene for prognosis and chemotherapy sensitivity. Methods: Forty HB patients were selected based on primary diagnosis and treated from January 2015 to May 2018 in Tongren Hospital Affiliated to Capital Medical University. The expression level of INI1 gene was detected by using fluorescence immuno-PCR. Clinical data of children with HB were collected, including staging, grouping, risk, efficacy and prognosis of chemotherapy. The relationship between the mRNA expression of IN1 gene and clinical data was analyzed by SPSS 21.0 software. Results: (1)Clinical characters: the medium age of 40 HB patients was(30.50±2.39) months, male 29 cases (72.5%), and female 11 cases (27.5%). There was 1 case (2.5%) of stage Ⅱ HB, 13 cases (32.5%) of stage Ⅲ HB, and 26 cases (65.0%) of stage Ⅳ. The low-and intermediate- risk group had 12 cases (30.0%) of HB, and the high-risk group had 28 cases (70.0%) of HB.(2)Clinical efficacy and prognosis: by July 1^st, 2018, the medium following-up time was (12.2±10.1) months, 4 patients (10%) were treated by chemotherapy without surgery, and 36 patients (90%) were treated by surgery .All patients (100%) received chemotherapy.The average cycle of chemotherapy lasted(13.17±0.02). Ten patients were dead and 30 patients survived till following-up.The overall survival (OS) rates were 75% and the event-free survival rates were 50% (20 patients with HB). (3) IN1 gene mRNA expression: The average PCR quantitative of IN1 gene mRNA of 40 HB patients(2^-ΔCT)was 0.31±0.70.The average mRNA quantitative of HB patients (2^-ΔCT)with high-risk and low/intermediate-risk group was 0.23±0.43 and 0.48±1.13(t=6.363, P=0.05). According to histology diagnosis, the average mRNA quantitative of small cell undifferentiated (SCU) type (12 cases)and other type (25 cases) was 0.09±0.11 and 0.43±0.86(t=4.533, P=0.04). The average mRNA quantitative of patients with or without radical surgery was 0.04±0.03 and 0.34±0.74 (t=2.935, P=0.022). The data of children with poor chemotherapy sensitivity(19 cases)and those sensitive to chemotherapy (21 cases) were 0.03±0.04 and 0.30±0.82, and the difference was statistically significant (t=5.688, P=0.018). Conclusions Poor therapeutic effect results in low mRNA expression of IN1 gene in children of HB.The IN1 gene expression could be an early warning factor for treatment sensitivity and prognosis.

Objective:The vasculature and canal were located using radiography after the fresh osseous specimens were decalcified, after which the anatomic investigation of intraosseous vasculature was conducted based on the orientation of the canals.Methods:To investigate the basic dissected methods for intraosseous vasculature and the related clinical significance. Methods The materials were obtained from seven fresh knee joint specimens from patients with amputation due to car accidents, nine fresh knee joint specimens from patients with amputation due to oncological radical surgery, and 44 knee joint specimens from 24 cadavers. Among them, 22 were males (55%) and 18 were females (45%), 28 were left knees (46.7%) and 32 were right knees (53.3%). 10 were aged from 16-90 years old (from 8 donors) and 50 were aged from 15-85 years old (from 32 donors). The tributaries of middle genicular vein which penetrate into the proximal tibial epiphysis and metaphysis via our previously discovered and denominated "foramen of tibial intercondylar eminence (FTIE)" were dissected as an example. After obtaining the fresh knee joint specimen, angiography was performed to observe the continuous extraosseous and intraosseous blood vessels. The first group of specimens with the removal of cortical bone was reserved in formalin solution at 4 °C for 7 d, sequentially immersed in Ethylene Diamine Tetraacetic Acid (EDTA), the decalcification agent, for 30 d with replacement for each two days. Based on the CT scanning and three-dimensional reconstruction, the orientation of bony canal which enclosed the vasculature was exposed to guide the anatomic incision. The exquisite dissection was achieved with the help of ophthalmological microsurgical instruments. The anatomical dissection were intuitively observed, compared with the angiographic images, and verified by histological examinations. The second group of samples was decalcified with strong acid as another strategy, and the comparison between different groups was conducted. To estimate the advantages and disadvantages of the two decalcification and dissection methods, and the distribution and universality of specific intraosseous vasculatures and canals, the methods can be utilized to dissect the diameter of the intraosseous vessels. Based on the anatomical study of intraosseous vasculature, the mechanisms including etiology, recurrence and spread of bone tumors and epiphyseal injuries were analyzed to improve the therapeutic regimen.Results:The intraosseous tributaries of middle genicular vein which penetrate into the tibial intercondylar eminence from the articular cavity were dissected, these vessels extended to the tibial metaphysis from epiphysis through the epiphyseal line or senescent physes. The diameter of the vessel entering the FTIE was 1.2 mm, and the intraosseous vessels divided into several tinier tributaries with the diameter of 0.3 mm to cross the epiphyseal line or closed physeal plate and differentiated into capillaries in the distal regions, therefore was difficult to dissect directly. The histological examinations confirmed the authenticity of intraosseous vessels. Compared with the samples decalcified with strong acid, the blood vessels were obviously dissolved, and only a few residual epithelial cells were observed under the light microscope. Based on the anatomical study of intraosseous vessels, the treatment protocols for some related bone tumors and epiphyseal injuries were modified and satisfactory results were achieved.Conclusion:The methods can realize the ideal direct dissection for the intraosseous blood vessels with the outer diameter greater than or equal to 0.3 mm.

Objective:To investigate the clinical characteristics of E. coli peritoneal dialysis-associated peritonitis (PDAP) and the risk factors for its occurrence and treatment failure.Methods:The clinical data of patients with episodes of PDAP in four general hospitals in Jilin Province from 2013 to 2019 were retrospectively reviewed. According to the pathogenic bacteria, the patients were divided into E. coli and non- E. coli groups. The incidence of E. coli PDAP in the last seven years was calculated and the clinical characteristics were compared between two PDAP groups. Logistic regression was used to analyze the risk factors for the occurrence and treatment failure of E. coli PDAP. Results:A total of 693 PDAP episodes/cases were enrolled in this study, including 100 episodes/cases in the E. coli group and 593 episodes/cases in the non- E. coli group. The incidence rate of E. coli PDAP in the four hospitals showed a decreasing trend during 2013 to 2019. Compared with the non-E.coli group, the proportion of diabetic patients and the average blood albumin levels in the E. coli group were lower ( χ2=5.006, Z=-2.992, P<0.05), while the proportion of refractory peritonitis was higher, and the duration of antibiotic therapy was longer ( χ2=6.350, Z=-2.779, P<0.05). Multivariate Logistic regression analysis showed that history of PDAP ( OR=1.577, 95% CI: 1.015-2.448) and low baseline serum albumin level ( OR=0.958, 95% CI: 0.923-0.995) were independent risk factors for the development of E. coli PDAP, while concomitant diabetes was an independent protective factor for E. coli PDAP ( OR=0.538, 95% CI: 0.330-0.876). Moreover, long-term dialysis was an independent risk factor for treatment failure of E. coli PDAP ( OR=1.047, 95% CI: 1.018-1.076). Conclusion:The incidence rate of E. coli PDAP in study institutions has declined in the past 7 years, but the rate of refractory PDAP is still high. The history of PDAP and low blood albumin level are independent risk factors for the occurrence of E. coli PDAP, while concomitant diabetes is an independent protective factor for the occurrence of E. coli PDAP. Long-term dialysis is an independent risk factor for treatment failure of E. coli PDAP.