1.Screening and identification of aptamers against the recombinant transpeptidase domain of PBP 2a
Xiaohua TANG ; Weiqin YANG ; Jiezhen HUANG ; Jianqiong HUANG
Chongqing Medicine 2016;45(17):2385-2388
Objective To screen and identify the aptamers of the recombinant transpeptidase domain of PBP2a(penicillin binding protein 2a ,PBP2a) .Methods By using the recombinant transpeptidase domain of PBP2a as the screening target ,oligonucle-otides which were capable of specifically binding to the protein were screened by a random oligonucleotide library through the stem -atic evolution of ligand by exponential enrichment (SELEX )technique .The ssDNA was cloned and sequenced ,and the secondary structure of aptamer clones was predicted with mfold program .Results After 11 cycles of the selection ,the aptamers which were capable of binding to PBP2a with high affinity have been selected .40 clones from the 8 and 10 cycles were selected randomly and se-quenced .The aptamers obtained had no obvious homology according to their sequences by the sequence alignments ,and the 40 aptamers were classified to three groups according to their secondary structures .The aptamer 13 was found to be specific for the target protein with the highest affinity .Conclusion Aptamers for the recombinant transpeptidase domain of PBP2a with high affili-ty and specificity were successfully screened by SELEX ,which lays a foundation for exploring new ways of diagnosis and treatment of MRSA infection .
2.Pathogen profile of bloodstream infections in low birth weight preterm infants:a report of 95 cases
Xiaohua TANG ; Xicai TANG ; Weiqin YANG ; Jiezhen HUANG ; Zihao OU
Chinese Journal of Infection and Chemotherapy 2015;(5):439-442
Objective To study the etiology and antibiotic resistance of bloodstream infections in low birth weight preterm infants .Methods A total of 95 cases of bloodstream infections in low birth weight preterm infants were treated in our hospital from January 2011 to April 2014 .The clinical data of these patients were analyzed retrospectively .Results A total of 96 pathogens were isolated ,including 57 strains of gram‐negative bacilli ,38 strains of gram‐positive cocci ,and 1 strains of Trichosporon asahii .The most frequently isolated pathogens were Klebsiella pneumoniae (40 strains)and coagulase‐negative Staphylococcus(31 strains).All gram‐negative bacilli were sensitive to carbapenems such as imipenem and panipenem . Streptococcus isolates were sensitive to most antibiotics .Most Staphylococcus isolates were methicillin‐resistant ,which were highly resistant to common antibiotics but all sensitive to linezolid , vancomycin and teicoplanin . Conclusions The most important pathogens responsible for bloodstream infections in low birth weight preterm infants in our hospital are K lebsiella pneumoniae and coagulase‐negative Staphylococcus . Early identification of responsible pathogen and rational antimicrobial therapy are critical for good prognosis of bloodstream infections in low birth weight preterm infants .
3.Clinicopathological features of gastric alpha-fetoprotein-producing adenocarcinoma with SWI/SNF complex deletion
Jiezhen YANG ; Xinxin GUO ; Chen XU ; Yingyong HOU ; Wei YUAN
Chinese Journal of Pathology 2024;53(1):52-57
Objective:To investigate the clinicopathological features and treatment of gastric alpha-fetoprotein (AFP)-producing adenocarcinoma with SWI/SNF complex deletion.Methods:Four cases of gastric AFP-producing adenocarcinoma with SWI/SNF complex deletion diagnosed in Zhongshan Hospital of Fudan University from January 2021 to December 2022 were collected, and their histomorphological characteristics, immunohistochemical (IHC), in situ hybridization of Epstein-Barr virus-encoded RNA (EBER), next-generation sequencing results, clinicopathological features and treatment were summarized, and literature review was conducted.Results:Among the 4 patients, there were three males and one female. They presented with abdominal pain, belching and melena. Serum AFP was significantly elevated in three patients, and endoscopy showed ulcerative lesions. Microscopically, the tumor cells showed mainly diffuse flaky or nest-like growth and typical characteristics of hepatoid adenocarcinoma. In two cases there were adenoid growth, and the tumor cells in these areas possessed clear cytoplasm, suggesting enteroblastic differentiation. The tumor cell nuclei were pleomorphic with large nucleoli and brisk mitoses. The IHC results showed that the tumor cells expressed AFP, GPC3 and SALL4, and there was retained expression of broad-spectrum keratin (CKpan) and E-cadherin. IHC detection of SWI/SNF complex subunits, namely INI1 (SMARCB1), BRG1 (SMARCA4), BRM (SMARCA2), ARID1A protein was performed. In all four cases the hepatoid adenocarcinoma region and enteroblastic differentiation region showed SMARCA2 deletion, and one case with enteroblastic differentiation also showed ARID1A deletion. SMARCB1 and SMARCA4 deletions were not seen. All the four cases were diffusely positive for p53 protein, and the Ki-67 proliferation index was 80%-90%. There were no mismatch repair deletion detected; one cases showed HER2 was strongly positive (3+), and EBER was negative. None of the four cases had mutations in the SWI/SNF complex-related subunits detected by next-generation sequencing. Among the four patients, two underwent palliative surgery due to distant metastasis at the time of surgery, two underwent radical resection. Postoperative adjuvant chemotherapy was given to three patients.Conclusions:AFP-producing adenocarcinoma is a rare subtype of gastric cancer, which can be combined with SWI/SNF complex deletion, and the pathomorphological manifestations are different from the classical SWI/SNF complex deletion of undifferentiated carcinoma with rhabdoid phenotype.
4.Effect of quality control circle on postoperative ambulation in patients with lumbar spinal stenosis
Yaoying? JIANG ; Lingyun LI ; Jiezhen LIU ; Fang WANG ; Minqi WU ; Ying YANG ; Bin WEI ; Yingjie LIANG ; Hongbin JU
Chinese Journal of Modern Nursing 2015;(19):2299-2303,2304
Objective To explore the effect of quality control circle ( QCC ) on postoperative ambulation among patients with lumbar spinal stenosis. Methods The QCC group had been established. 5M selection method was used for the topic “To improve the correct rate to get out of bed for postoperative patients with lumbar spinal stenosis”, and then we made plan time table, set up the goal-89. 44% correct rate o get out of bed for postoperative patients with lumbar spinal stenosis after analyzing the incorrect reasons, confirmed 5 reasons after argument, formulated countermeasure and checked out the effects. Results After QCC activities, the rate of correct ambulation ( get out the bed ) amounted 92%, while 76% before the activities, with a significant difference (χ2 =9. 52,P<0. 01). Level of specialty, working passion, team spirit, ability of QCC activity, problem solving skills and communication skills had been improved. Conclusions QCC activities can significantly improve the correct rate of getting out bed, effectively shorten the average length of hospitalization, and improve patients′satisfaction and professional level in clinical nursing.
5.Therapeutic effects of high monounsaturated fatty acid and low carbohydrate formula on blood glucose levels and diarrhea in critically ill neurological patients
Xiaochang HUANG ; Rong LAI ; Qiuliang YANG ; Jiezhen FENG ; Yongjing SU ; Huiyu FENG ; Hongyan ZHOU
Chinese Critical Care Medicine 2024;36(9):980-984
Objective:To investigate the effects of using a high monounsaturated fatty acid (MUFA) and low carbohydrate formula on blood glucose levels and diarrhea treatment effects in critically ill neurological patients.Methods:A self-controlled before-and-after study design was employed, with 13 patients admitted to the neurology intensive care unit (ICU) of the First Affiliated Hospital of Sun Yat-sen University from November to December 2023, who were treated with a high MUFA and low carbohydrate formula [Glucerna enteral nutrition (EN) preparation]. Changes in blood glucose parameters within 7 days before and after the use of Glucerna EN preparation were analyzed, including standard deviation ( SD) of blood glucose, mean blood glucose (MG), median blood glucose, mean amplitude of glycemic excursions (MAGE), largest amplitude of glycemic excursions (LAGE), coefficient of variation ( CV) of blood glucose, the incidence of hyperglycemia (> 7.8 mmol/L) and severe hyperglycemia (> 13.9 mmol/L), and daily insulin dose. Changes in total protein (TP), albumin (ALB), hemoglobin (Hb), C-reactive protein (CRP), and white blood cell count (WBC) were observed before and after intervention. Improvement in diarrhea symptoms, Hart diarrhea score, Bristol Stool classification score, and incontinence dermatitis classification were also analyzed before and after the use of Glucerna EN preparation. Results:A total of 13 critically ill neurological patients were enrolled, among whom 9 patients had a history of hyperglycemia and 8 patients had diarrhea symptoms. After intervention with Glucerna, the patients' SD of blood glucose, MG, median blood glucose, MAGE, LAGE, CV of blood glucose, incidence of hyperglycemia, incidence of severe hyperglycemia, and daily insulin dose were all lower than those before the intervention [ SD of blood glucose (mmol/L): 1.83±1.11 vs. 2.10±1.13, MG (mmol/L): 8.87±2.03 vs. 9.75±1.37, median blood glucose (mmol/L): 9.12±1.67 vs. 10.17±0.48, MAGE (mmol/L): 0.66±0.31 vs. 0.78±0.32, LAGE (mmol/L): 4.95±3.64 vs. 5.58±3.10, CV of blood glucose: 16.00% (11.00%, 28.50%) vs. 18.00% (12.50%, 27.50%), hyperglycemia incidence: 47.31% vs. 74.66%, severe hyperglycemia incidence: 6.08% vs. 6.71%, daily insulin dose (U): 5.25 (0.00, 32.59) vs. 20.76 (0.00, 66.88)], with a significant decrease in daily insulin dose after the intervention ( P < 0.05); TP, ALB, Hb, CRP and WBC showed no significant changes before and after the intervention with Glucerna EN preparation. The improvement time of diarrhea symptoms after intervention was (3.50±1.41) days, and the Hart diarrhea score on the seventh day after intervention (4.88±3.48 vs. 10.00±3.38) and the Bristol Stool classification score on the third and seventh days after intervention (5.87±0.35, 5.50±0.53 vs. 6.50±0.53) were significantly lower than before the intervention (all P < 0.05). Before the intervention with Glucerna EN preparation, the classification of incontinence dermatitis was mainly classified as Grade 2 severity (71.43%); after the intervention, it significantly improved by the seventh day, with Grade 1 being the main classification (57.14%). Conclusion:The high MUFA and low carbohydrate formula has a positive effect on blood glucose control and diarrhea treatment in critically ill neurological patients.
6.Mechanism of liposome curcumol reversing cisplatin resistance in ovarian cancer
Jiezhen Yang ; Jing Wang ; Yonghong Song ; Qin Tang ; Qiang Wu
Acta Universitatis Medicinalis Anhui 2022;57(7):1106-1111
Objective:
To investigate the effect and mechanism of liposome curcumol( LC) on cisplatin resistance of human ovarian cancer cells.
Methods:
Ovarian cancer cisplatin resistant cell line SKOV3/DDP was used. Cells were divided into five groups: blank control group,negative control group( containing 12. 5 μg/ml liposome),cisplatin group( 4 μg/ml),LC group( 20 μg/ml),and combined group( cisplatin( 4 μg/ml) + LC( 20 μg/ml).Cell proliferation was detected by CCK8 assay,the content of cisplatin in cells was detected by high performance liquid chromatography-tandem mass spectrometry( HPLC-MS),and apoptosis was detected by flow cytometry. The level of P-glycoprotein( P-gp) mRNA was detected by qRT-PCR and the expression of P-gp protein was detected by immunohistochemistry and Western blot.
Results:
The cell proliferation ability of the combination group was the lowest while the apoptosis rate was the highest( P<0. 05) among the groups. HPLC-MS showed that the content of cisplatin in combined group was higher than that in cisplatin group( P<0. 05). P-gp mRNA and protein in the combined group were down-regulated( P<0. 05).
Conclusion
LC can reduce cisplatin resistance of human ovarian cancer cell line SKOV3/DDP by inhibiting the expression of P-gp.