1.Establishment and preliminary application of a real-time fluorescent quantitative PCR assay for detection of murine norovirus
Chinese Journal of Comparative Medicine 2016;26(12):70-76
Objective To establish a real-time fluorescent quantitative PCR ( FQ-PCR ) method for detection of murine norovirus ( MNV) in laboratory mouse and provide the basis for establishment of a standard detection method for MNV.Methods Specific primers were designed and MNV DNA standards were prepared according to the MNV genome sequences published on NCBI .The specificity, sensitivity, repeatability and stability of the established Q-PCR method were tested.The established Q-PCR method was applied to detect 766 mouse caecum content samples to explore preliminarily the infection status of laboratory mice in Beijing .Results No cross reaction showed in human norovirus and feline calicivirus with the established Q-PCR method.The sensitivity was up to 10 copies/μL.The coefficient of variation ( CV) of intra-assay and inter-assay was less than 2%.There were 301 positive cases detected in the 766 samples of laboratory mice.Conclusions The established FQ-PCR method is accurate and effective with high specificity , sensitivity and repeatabiliy in the quantitative detetion of nucleic acid , and can be applied to rapidly and quantitatively screen MNV in laboratory mice .
3.Study on Activation of Nuclear Factor Erythroid 2-related Factor 2 Pathway with Fructus Schisandrae Extracts Medicated by Kinase Pathways Such as Extracellular Signal-regulated Kinase
Yang YU ; Qiao LAI ; Changqiang HE ; Jinlian HE ; Jie GAO
Journal of Guangzhou University of Traditional Chinese Medicine 2014;(6):957-962
Objective To observe the influence of several kinase pathways such as extracellular signal-regulated kinase ( ERK) , and mitogen-activated protein kinase p38 ( p38MAPK) on nuclear factor erythroid 2-related factor 2 (Nrf2) pathway activated by Fructus Schisandrae extracts (FSE) . Methods HepG2 cells were treated by FSE for 24 hours after pretreatment with protein kinase inhibitors for 2 hours. The mRNA expression levels of Nrf2 and downstream target genes heme oxygenase-1 (HO-1), NAD (P) H quinine oxidoreductase 1(NQO1), P-glycoprotein ( P-gp) and multidrug resistance-associated protein 2 ( MRP2) were detected by real-time polymerase chain reaction ( RT-PCR) , and their protein expression levels and Nrf2 nuclear translocation were measured by Western blotting method. Results RT-PCR results showed that the mRNA expression levels of HO-1, NQO1, P-gp and MRP2 activated by FSE in HepG2 cells were inhibited by PD98059, SB203580 and Rottlerin, and the mRNA expression of Nrf2 was suppressed only by SB203580 and Rottlerin. Western blotting results showed that the mRNA expression levels of HO-1 and P-gp activated by SCE in HepG2 cells were inhibited by PD98059, SB203580 and Rottlerin. In addition, the protein expression of Nrf2 in HepG2 cytoplasm was increased by the above three inhibitors, and nuclear translocation of Nrf2 was inhibited by PD98059 and SB203580. Conclusion The mechanism of FSE activating Nrf2 pathway may be associated with the increase of Nrf2 nuclear translocation through the direct phosphorylation of Nrf2 induced by ERK and p38MAPK.
5.A comparison between adults and children tonsillectomy with monopolar electrocautery.
Min AO ; Jie DENG ; Lei GAO ; Gang HE
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2015;29(3):240-242
OBJECTIVE:
Tonsillectomy is one of the most frequently applied operations in the ENT practice. This prospective study compared intraoperative records and postoperative clinical outcomes between adults and children patients following monopolar electrocautery tonsillectomy.
METHOD:
Forty adult patients and Forty children patients with histories of recurrent tonsillitis or hypertrophic tonsillitis were enrolled. Intraoperative parameters and postoperative outcomes were compared.
RESULT:
Children tonsillectomy with monopolar electrocautery was significantly faster to perform (P < 0.05), and produced significantly less intraoperative blood loss (P < 0.05), and faster to return to commencement of a regular diet (P < 0.05) than adults. Children tonsillectomy endured less postopera- tive pain within a week (P > 0.05) than adults, but there was no significant difference in pain on the 14th postoperative day in two groups. There was no obvious postoperative hemorrhage in two groups. There was no significant difference in postoperative tonsillar fossa healing and postoperative temperature between the groups.
CONCLUSION
Children and adults tonsillectomy with monopolar electrocautery had clinical characteristics respectively. Monopolar electrocautery tonsillectomy was safe and operated easily in both two groups.
Adult
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Blood Loss, Surgical
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Child
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Electrocoagulation
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Humans
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Pain, Postoperative
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Postoperative Hemorrhage
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Postoperative Period
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Prospective Studies
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Tonsillectomy
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methods
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Tonsillitis
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surgery
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Wound Healing
6.Clinical application of tacrolimus in children with primary nephrotic syndrome
Zhengkun XIA ; Guangling LIU ; Yuanfu GAO ; Zhongmin FAN ; Jie FU ; Yuanfeng FU ; Xu HE
Chinese Journal of Nephrology 2009;25(3):187-190
ObjectiveTo investigate the clinical application of tacrulimus (TAC, FK506) in children with primary nephrotic syndrome (NS). MethodsSixty-five primary NS children received routine or decreased-dosage glucocorticosteroid according to clinical NS types after hospitalization. At the same time, TAC was given orally with the dosage of 0.1 to 0.15 mg/kg, once every 12 hours, for 6 to 24 months. And the serum concentration of TAC was monitored during the course. ResultsAfter the treatment of TAC for 1 to 2 months, 65 patients were recovered with gradually reduced urinary protein, rapidly increased serum albumin, and improvement of cholesterol and triglycerides. Total remission rate was 83.1% and onset time was 7 to 54 days. Twelve cases experienced recurrence. Increased CD4, as well as 3/3 or 3/1 TAC genotype, indicated higher remission rate. Various pathological types had different remission rates or ratio, which were as follows: minimal change nephropathy (96.4%), mesangial proliferative glomendonephritis (90.0%), membranous nephropathy (2/3), membranous proliferative glomerulonephritis (3/5), focal segmental glomerulosclerosis (4/9). The patients would recover in the course of treatment under the conditions of TAC initial dose as 0.1 to 0.15 mg /kg per 12 hours and controlled serum concentration as 5 to 10 g/L. During the treatment, 12 cases appeared gastrointestinal symptoms, mainly as anorexia, nausea and vomiting, 1 abdominal pain, 2 headache, 1 tremor, 1 paresthesia, 3 insomnia, 4 transient increased Scr, 8 slightly increased NAG, 6 increased C3 and α-2 macroglobulin. The symptoms disappeared within one week or after stopping TAC. ConclusionsTAC is effective in primary NS children, even with abnormal liver function or tuberculosis infection. TAC can also be a substitute to cyclosporine A.
7.Characteristics of coronary arteriography and traditional Chinese medicine syndrome of 1,069 patients with coronary artery disease
Jie WANG ; Yanwei XING ; Jianxin CHEN ; Qingyong HE ; Yonghong GAO ; Zun LI
Journal of Integrative Medicine 2008;6(2):148-52
OBJECTIVE: To study the characteristics of coronary arteriography and traditional Chinese medicine syndrome of 1,069 patients with coronary artery disease (CAD). METHODS: One thousand and sixty-nine patients with CAD were investigated by epidemiological method. The patients were divided into young patients (n=82, aged 45 years or younger) and middle-aged and old patients (n=987, older than 45 years). The characteristics of the two groups were analyzed, including clinical data, coronary arteriography and traditional Chinese medicine syndrome. RESULTS: Compared with middle-aged and old patients, proportion of male, triglyceride, total cholesterol, smoking patients, acute myocardial infarction and family history of CAD in young patients were significantly higher (P<0.05). Patients accompanying with hypertension and diabetes in middle-aged and old patients were more than those in young patients (P<0.05). Occurrence rates of morbidity of left circumflex coronary artery, left main coronary artery and multi-branch were higher in middle-aged and old patients (P<0.05), however, the occurrence rates of morbidity of single and double-branch were higher in young patients (P<0.05). The occurrence rates of syndromes of qi stagnation and phlegm turbidity in young patients were higher than those in middle-aged and old patients (P<0.05). But the proportions of cold coagulation, yin deficiency, yang deficiency and kidney deficiency in middle-aged and old patients were obviously higher (P<0.05). CONCLUSION: The traditional Chinese medicine syndrome and pathological changes of CAD in young patients are different from those in old patients.
8.Pathogenic bacteria distribution and drug susceptibility characteristics of the patients with lactation acute mammitis
Zhiqing YAN ; Xiangjun MA ; Qing WANG ; Jie WANG ; Haifeng GAO ; Xiangping HE ; Yi ZHANG
Clinical Medicine of China 2014;(7):732-734
Objective To explore the pathogenic bacteria species of the patients with lactation acute mammitis and their sensitivities to antibiotics in order to provide a guideline in choosing antibiotics reasonably. Methods Four hundred and thirty-three samples from diseased breast(include milk,puncture fluid and secretion in ulcerateing skin)from 310 patients with lactation acute mammitis,who were treated by the center of mammary gland of Maternal and Child Health Care of Haidian district in BeiJing from Jan. to Aug. in 2012. Statistics analysis was made to analysis the pathogenic bacteria species and the characteristics of drug sensitivity. Results (1)Of the 433 samples,407 strains of pathogens were picked out,which consisted of 215 strains of Staphylococcus aureus(SAU),43 strains of methicillin-resistant staphylococcus aureus( MRSA),43 strains of coagulase negative staphylococcus(CNS),42 strains of Methicillin-resistant staphylococcus epidermis (MRSE),22 strains of Staphylococcus epidermidis(SE),12 strains of Methicillin-resistant coagulase negative staphylococci(MRScon)and 8 strains of Alpha hemolytic streptococcus.(2)There were 237 strains of G +bacteria,and the drug sensitive test showed that they were all sensitive to vancomycin(100% ),91. 8% were sensitive to penicillin,57. 0% to clindamycin( CLDM))and 69. 0% to erythrocin. Meanwhile,94. 0% were sensitive to levofloxacin,70. 0% to cephalosporins,and 80. 0% to gentamicin. In addition,there were 43 MRSA, and drug sensitivity test showed that 95. 0% were sensitive to levofloxacin,96. 0% to gentamicin,97. 0% to macrodantin,and 92. 0% to cotrimoxazole. Meanwhile,it's drug resistance rate to erythrocin was 13. 0% and 15. 0% to clindamycin( CLDM). MRSA was completely resistant to Lactam antibiotics( 100% ) such as penicillin and Cephalosporins,95. 0% to levofloxacin,96. 0% to gentamicin,97. 0% to furadantin and 92. 0% to bactrim. Conclusion The predominant pathogenic bacteria of lactation acute mammitis include SAU,MRSA, MRSA,CNS,MRSE,SE and they are all the G + bacteria. They are showed with a high rate of drug resistance, especially for the sensitive drugs which had been proved in the past. Due attention should be paid to germiculture from diseased breast and drug sensitive test as soon as possible,and to provide a guideline in choosing antibiotics reasonably.
9.Prenatal ultrasound diagnosis of fetal aortic atresia
Yan XU ; Jinghai HE ; Lianli JIE ; Jian CHENG ; Xiang GAO ; Qinghua LIU
Chinese Journal of Ultrasonography 2012;(11):946-948
Objective To investigate the clinical value of prenatal ultrasonography in diagnosing fetal aortic atresia.Methods Ultrasound images of 9 cases with fetal aortic atresia confirmed by autopsy were retrospectively analysed and summarized.Results Fetal echocardiography showed ascending aorta and aortic arch were thin in seven cases and unclear in two cases.Color Doppler flow imaging showed fetal aorta no blood connection with left ventricle and reverse blood flow in aortic arch among all 9 cases.There were 6 cases associated with hypoplastic left heart syndrome,two cases associated with ventricular septal defect and well-developed left ventricle,one case complicated with complete atrioventricular septal defect.Nine cases were detected at average gestational age (26.0 ± 2.6) weeks,minimum gestational age was only 16 weeks pregnant.Conclusions Prenatal ultrasound diagnosis of fetal aortic atresia has important clinical value.Ascending aorta and aortic arch are thin or unclear,no blood flow connection between fetal aorta and left ventricle and reverse blood flow in aortic arch are major diagnostic evidences.Color Doppler flow imaging is helpful for early diagnosis.
10.Development of the Diode method for patient's dose measurement in radiotherapy
Zhijian HE ; Jilong YUAN ; Lei GAO ; Suming LUO ; Jie QIU ; Bo YANG ; Tingtian PANG
Chinese Journal of Radiological Medicine and Protection 2011;31(3):355-358
Objective To explore the measurement method of the treatment dose of the patient with Diode for photon beam in radiotherapy,and to validate the treatment dose by comparing with the treatment planning system (TPS).Methods Experiments of the reproducibility,dose rate dependence,non-linearity dose response,and calibration factor in 60Co γ and 6 MV X beams were carried out with Diode on the surface of solid phantom and in water phantom.According to the needs of clinic treatment,different conditions were chosen to observe the dose changes with the angle of incidence,energy response,distance of source to skin,field size,wedge angle,block and tray using ionization chamber and water phantom.The Diode was placed on the surface of the solid phantom to obtain the correction factors.The doses of the chest,abdomen,and head and neek were verified with the Alderson phantom and Diode.Diode doses of the pelvis,head and neck at 14 points on the patient were measured.Results The Diode was irradiated at the points of the Alderson phantom,such as AP,RL and LL of the pelvis,with and without wedges,RL and LL junction of the neck and chin,with and without mask,the maximum relative deviation of doses was within ± 3% between Diode and TPS.The Diode was placed in different locations on the patient,including chest,abdomen and head and neck.The relative maximum deviation of doses was within ±5% between Diode and TPS.Conclusions The Diode method is reliable for measuring the exposure doses of the patient in radiotherapy.