1.Establishment and application of a Taqman MGB real-time PCR for the detection of Streptobacillus moniliformis
Jin XING ; Yufang FENG ; Bingfei YUE ; Zhengming HE ; Fangwei DAI ; Xiaoying SA ; Jiejie DAI
Chinese Journal of Comparative Medicine 2015;(8):62-67
Objective To establish a real-time quantitative PCR ( qPCR) method for detection of Streptobacillus moniliformis, which can be used to rapidly detect this pathogen in laboratory animals .Method According to the S. moniliformis sequences published in NCBI , we designed specific primers and MGB probe .The specificity, sensitivity and stability of this method were evaluated using 24 standard reference strains .Total of 823 respiratory specimens of animals including mice, rats, guinea pigs, hamsters, rabbits, Mongolian gerbils and tree shrews , were detected by this established Taqman MGB qPCR method .Results We had successfully established the S.moniliformis Taqman MGB qPCR method . S.moniliformis was not detected in the samples of mice , rats, guinea pigs, hamsters and rabbits.The positive rate of S. moniliformis was 1.5% ( 1/65 ) and 61.7% ( 37/60 ) in conventional Mongolian Gerbils and tree shrews , respectively . Conclusions Our developed qPCR method can be used to effectively detect S.moniliformis in laboratory animals .Moreover , its accuracy and sensitivity are better than the national standard method .This study laid the foundations for optimizing the quality inspection system of laboratory animals .
2.Dynamic study on levels of serum PRL, LH, FSH, E2 and development of ovarian follicles in different phases of estrus cycle in gerbils
Li XIAO ; Liqing CHEN ; Fangwei DAI ; Xiaoming SONG ; Honggang GUO ; Lingqun LU ; Yu LV ; Xiaoying SA
Chinese Journal of Comparative Medicine 2014;(6):30-34
Objective To investigate the numbers of corpus luteum and ovarian follicles and compare the levels of serum prolactin (PRL), luteinizing hormone (LH), follicle-stimulating hormone (FSH) and estradiol (E2 ) in different phases of estrus cycle in female gerbils .Methods Consecutively taking vaginal smears of the gerbils and directly examined under light microscope to distinguish the four phases of the estrus cycle .Hematoxylin and eosin staining was used to histological examination of the gerbil ovaries , and to detect the levels of serum PRL , LH, FSH and E2 by ELISA assay during estrus cycle .Results The proportion of cornified vaginal exfolliated cells could be the basis to distinguish four phases respectively:proestrus, oestrus, metoestrus, and dioestrus.Moreover, there were no significant differences between the numbers of ovarian follicles in different phases of estrus cycle .The numbers of corpus luteum in preoestrus were significantly lower than that in the other phases of estrus cycle ( P <0.05 ) .The levels of serum PRL and LH were increasing constantly from preoestrus to dioestrus , and both reached a peak at dioestrus ( P<0.05 ) .The levels of serum FSH and E2 both peaked at preoestrus , and were significantly higher than those at oestrus , metoestrus and dioestrus ( P<0.05).Conclusions There are no significant differences between the numbers of ovarian follicles in different phases of estrus cycle .Gonadotropin , prolactin and estradiol paly important roles in the regulation of estrous cycle .The phases during which surges of FSH and E 2 occur in Mongolian gerbils are similar to those of rats and mice , while the PRL and LH are different .Our findings provide further reference to the study of reproductive physiology of Mongolian gerbils .
3.Establishment and application of a fluorescence quantitative Taqman-PCR detection method of mouse poxvirus
Jiangtao DU ; Fangwei DAI ; Shasang ZHOU ; Xiaoming SONG ; Yu LV ; Xiaoying SA
Chinese Journal of Comparative Medicine 2015;(6):59-64
Objective To establish a fluorescence quantitative Taqman-PCR method for rapid and accurate detection of mouse poxvirus.Methods After sequence alignment and comparison, ERPV_027 gene was selected as the primer and probe design gene.Furthermore, the specificity, sensitivity, stability and reproducibility of these primers and probes were detected.Results The detection limitation of this method was 68 copies/μL.Data showed that this method has high specificity, which specifically amplifies mouse poxvirus, with no amplification signal of mouse hepatitis virus, Sendai virus, Salmonella and some other viruses and bacteria.This method also showed good stability and reproducibility. Conclusions This study has successfully established a fluorescence quantitative Taqman-PCR method for detection of mouse poxvirus, with high specificity, sensitivity, good stability and reproducibility, and a broad application potential.
4.Clinical efficacy of patient-controlled intravenous analgesia for intractable cancer pain
Jianguang LIN ; Tianwen XU ; Fangwei XIE ; Deqiang FU ; Yijun DAI ; Aiyue ZHAO
Chinese Journal of Clinical Oncology 2015;(12):586-589
Objective:To compare the clinical efficacy of oxycodone hydrochloride controlled-release tablets (OHCT) and pa-tient-controlled intravenous analgesia (PCIA) in the treatment of intractable cancer pain. Methods:Retrospective analysis was conduct-ed to evaluate the intractable cancer pain of 89 elderly patients who were admitted to the medical oncology departments of The Second Affiliated Hospital of Fujian Medical University and the Fuzhou General Hospital of Nanjing Military Command between September 2012 and March 2014. Among the 89 patients, 47 were treated with OHCT, and 42 received PCIA. The total dosage ranged from 60 mg/d to 400 mg/d PO q12h for patients in the OHCT group, whereas abackground dose+patient-controlled dosemode was adopted for patients in the PCIA group. The therapeutic efficacy, presence of adverse reactions, cost of treatment, and degree of patient satisfaction were compared between the two groups. Results:The average dosages of analgesics in the two groups were almost the same (P>0.05). Visual analogue scale (VAS) values and daily average VAS values were both lower in the PCIA group than in the OHCT group at 24 h after analgesia (P<0.05). The incidence of adverse reactions, such as nausea and vomiting, was also lower in the PCIA group than in the OHCT group (P<0.05). The cost of treatment and degree of patient satisfaction were the same in both groups (P>0.05). Conclusion:Pa-tients who received PCIA attained better analgesia and exhibited less adverse reactions than those who received OHCT whereas the treatment cost and patient satisfaction did not differ in both groups.
5.Histological Characteristics of the Kidney in Mongolian Gerbils of Different Ages
Lingqun LU ; Honggang GUO ; Qiaojuan SHI ; Fangwei DAI ; Xiaofeng CHU
Laboratory Animal and Comparative Medicine 2023;43(1):61-66
ObjectiveTo explore the histological characteristics of the kidney in Mongolian gerbils of different ages. MethodsTen Mongolian gerbils of 2, 6, and 12 months old (half male and half female) were selected. After euthanasia, the kidneys were taken and paraffin sections were made. After HE, MASSON and PAS staining, the structural differences of different parts of the kidney tissue in gerbils of different ages were observed by digital scanning, and the relevant data of the kidney tissue were measured by image analysis software. ResultsThe number of proximal convoluted tubules was more than that of distal convoluted tubules in the renal cortex and outer medulla of gerbils. With age, the glomerular density decreased, the glomerular diameter increased, the basement membrane of renal tubules thickened, and the fibrous components between renal tubules increased. ConclusionThe histological structure of Mongolian gerbil's kidney varies with age, which may be related to glomerulosclerosis and parenchymal cell reduction. The specific mechanism needs further study.