According to relevant national laws and regulations, practitioner training was included into laboratory animal science teaching reform.By adjusting the training content and teaching method and use of animal models of typical human diseases, the transformation of training mode was realized and improved.By the assessment of basic theory in combination with practical operation, the thinking ability and hands-on skill of the practitioners are much improved. Through classroom instruction, experimental teaching, quality assessment and tracking survey, the evaluating process of the training quality of training teaching is performed.Therefore, the teaching reform of the laboratory animal science based on the training of practitioners is established.

Objective To investigate the relationship between truncating mutations of APC gene and sporadic colorectal cancer,and analyze the feasibility of non-radioactive protein truncation test (PTT) in the detection of mutations of APC gene.Methods Ninety-six samples of sporadic colorectal cancer tissues ( including 44 patients with colonic cancer and 52 patients with rectal cancer) were obtained from the General Hospital of Ningxia Medical University from September 2008 to September 2010.The mutation cluster region of the APC gene was screened using digital PTT labeled with fluorescent Lys-t-RNA,with a polymerase chain reaction fragment amplified from genomic DNA serving as a tenplate for in vitro translation.The occurrence of gene mutation was determined according to the emergence of truncated peptides.The mutation cluster region of the APC gene in 46 samples of colorectal cancer tissues was analyzed by direct sequencing.The detection rates of the 2 methods were compared by chi-square test.Results Thirteen (26％) truncated peptides were detected in the 50 samples of colorectal cancer tissues.The mutation type of 4 samples is nonsense mutation,which resulted in emergence of truncated gene products.Eleven (24％) truncated peptides were detected in the 46 samples of colorectal cancer tissues.There was no significant difference in the detection rates between PTT and direct sequencing ( x2 =0.033,P ＞ 0.05 ).Conclusions Truncating mutations of APC gene are common alterations in sporadic colorectal cancer in the Chinese.Digital PTT labeled with fluorescent Lys-t-RNA is rapid and high-sensitive in screening gene mutations.

Objective To investigate the effect of intraspinal grafting of brain derived neurotrophic factor (BDNF) ex vivo transgene myoblasts cells and methylprednisolone on caspase-3 expression after spinal cord injury (SCI). Methods A total of 120 experimental rats were divided into Group A (spinal cord contusion injury group), Group B (grafting of BDNF ex vivo transgene myoblasts cells group), Group C (methylprednisolone intravenous injection group) and Group D (grafting of BDNF ex vivo transgene myoblasts cells and methylprednisolone intravenous injection group). At days 1, 3, 7, 14 and 28 respectively after SCI, the expression of caspase-3 was measured immunohistochemically for quantitative analysis via a computer image analysis system. The motion functional recovery of the rats was observed by praxiologic and electrophysiologic examination. Results Positive expression cells of caspase-3 were found in all groups, with number from the highest to the lowest in order of Group A, Group B, Group C and Group D (P

Adolescent ; Adult ; Anesthesia, Intravenous ; Anesthetics, Intravenous ; Cardiopulmonary Bypass ; Female ; Fentanyl ; Heart Valve Prosthesis Implantation ; Humans ; Male ; Middle Aged ; Sufentanil

Objective To investigate the effect of dexamethasone on the pulmonary gas exchange in patients undergoing cardiac valve replacement under cardiopulmonary bypass(CPB).Methods Forty ASAⅡorⅢpatients aged 29-47 yrs weighing 50-69 kg undergoing cardiac valve replacement under CPB were randomly divided into 2 groups(n=20 each):dexamethasone group received dexamethasone 0.5 mg?kg~(-1) after induction of anesthesia and control group received normal saline(NS).Blood samples were taken before operation(T_0) immediately before CPB(T_1)and immediately after discontinuation of CPB(T_2)for determination of plasma total and active matrix metallo-proteinase-9(MMP-9)concentration(by enzyme-linked immuno-absorbent and fluorometric enzyme-linked immuno-absorbent assay respectively)and MMP-9 gene expression(RT-PCR).Blood samples were taken from radial artery at T_1 and T_2 for blood gas analysis.A-aDO_2 was calculated.Results MMP-9 gene expression and plasma total and active MMP-9 concentrations were significantly increased at T_2 as compared with those at T_0 in both groups and were significantly lower in dexamethasone group than in control group(P＜0.05 or 0.01).The A-aDO_2 at T_2 was significantly smaller in dexamethasone group than in control group.Conclusion Dexamethasone can inhibit the increase in gene expression,protein synthesis and activation of MMP-9 and decrease in gas exchange across alveolar-capillary membrane caused by CPB and protect the lungs during open heart surgery performed under CPB.

Objective To assess the value of MSCT and postprocessing techniques in displaying abnormal blood vessels of pulmonary sequestration. Methods Ten patients with pulmonary sequestration proved with operation and pathology underwent both plain and enhanced chest CT scan. The reconstructed images of maximum intensity projection, multiplanar reconstruction and volume rendering were analyzed retrospectively. Results Of 10 patients, intra-lobe type sequestration was seen in 9 patients and extra-lobar type in 1 patient. The lesions located in the left lower lobe in 8 patients and in the right lower lobe in 2 patients. Angiography postprocessing technique displayed abnormal supplying vessels in 9 patients (7 from descending aorta and 2 from abdominal aorta) and displayed the origin, courser, branches and distribution of lesions clearly. Vessels in 9 patients drained into inferior pulmonary veins except in one drained into azygos vein. Conclusion MSCT angiography with postprocessing reconstruction techniques can display abnormalities of pulmonary sequestration clearly and has great value in the diagnosis of pulmonary sequestration, so as might be the first choice in diagnosing pulmonary sequestration.

[ABSTRACT]AIM:Toinvestigatetheeffectofhypoxiacombinedwithexercisetrainingonmitochondrialcon-tent, and the role of mitochondrial biogenesis and mitophagy in this process .METHODS:Male Sprague-Dawley rats were randomly divided into 4 groups:normoxia control (NC) group, normoxia+training (NT) group, hypoxia+control (HC) group, and hypoxia+training (HT) group.The hypoxic animals were housed in normobaric hypoxic tent (11.3 % oxy-gen) for consecutive 4 weeks.The exercise training animals were exercised on a motor-driven rodent treadmill (5°) at a speed of 15 m/min, 60 min/d, 5 d/week for 4 weeks.Mitochondrial membrane potential was determined using JC-1 fluo-rescent probe .ATP synthesis capacity was determined using a bioluminescence technique .The protein expression of cyto-chrome C oxidase IV (COXIV), voltage-dependent anion channel-1 (VDAC-1), peroxisome proliferator-activated receptor gamma cofactor 1 alpha (PGC-1α), mitochondrial transcription factor A (Tfam),Bcl-2/adenovirus E1B 19 kD-interacting protein 3 (Bnip3) and beclin-1 in the muscles was detected by Western blotting .RESULTS:Compared with NC group, hypoxia attenuated mitochondrial membrane potential , ATP synthesis capacity , and the expression of COXIV , VDAC-1, PGC-1αand Tfam.Furthermore, hypoxia increased the expression of Bnip 3 and beclin-1.Compared with HC group , the exercise training elevated mitochondrial membrane potential , ATP synthesis capacity , and the expression of COXIV , VDAC-1, PGC-1α, Tfam, Bnip3 and beclin-1.CONCLUSION:A combination of reduced mitochondrial biogenesis and increased mitophagy seems to be responsible for the decrease in mitochondrial content after hypoxia .Exercise training with hypoxia elevates mitochondrial content and function in hypoxia , which may be mediated by appropriate increase and co-reg-ulation of mitochondrial biogenesis and mitophagy .

Objective:To observe the effect of warm-unblocking acupuncture plus fluticasone propionate nasal spray on the pulmonary ventilation, level of interferon-γ (IFN-γ) and sleep quality in patients with allergic rhinitis (AR). Methods: A total of 112 AR patients were enrolled between January 2013 and August 2018 and were divided into an observation group and a control group by the random number table method, with 56 cases in each group. Patients in the observation group received warm-unblocking acupuncture plus fluticasone propionate nasal spray, and patients in the control group only received fluticasone propionate nasal spray. The nasal symptom score, pulmonary function indexes, the levels of IFN-γ and interleukin (IL)-4 in serum, and sleep quality in the two groups were compared. Results: After treatment, the total effective rate in the observation group was higher than that in the control group (P＜0.05). The nasal symptom score dropped in both groups after treatment (both P＜0.05), and the score in the observation group was lower than that in the control group (P＜0.05). The pulmonary ventilation indexes all increased significantly after treatment in the observation group (all P＜0.05); the forced expiratory volume in 1 second (FEV1) to forced vital capacity (FVC) ratio (FEV1/FVC) and the forced expiratory flow at 50%, 75% and 25%-75% of the vital capacity (FEF50%, FEF75%, FEF25%-75%) increased after treatment in the control group (all P＜0.05); the pulmonary ventilation indexes were higher in the observation group than those in the control group (all P＜0.05). The level of IFN-γ increased significantly after treatment in the two groups (both P＜0.05) and the level of IL-4 dropped significantly (both P＜0.05); the observation group had a higher IFN-γ level (P＜0.05) and a lower IL-4 level (P＜0.05) compared with the control group. Regarding the Pittsburgh sleep quality index (PSQI), the scores of subjective sleep quality, habitual sleep efficiency and sleep disturbances and the general PSQI score decreased significantly after treatment in both groups (all P＜0.05), and the scores in the observation group were significantly lower than those in the control group (all P＜0.05). Conclusion: Warm-unblocking acupuncture plus fluticasone propionate nasal spray can effectively control the clinical symptoms and improve pulmonary function in the treatment of AR; this approach can regulate the levels of IFN-γ and IL-4 towards the normal range in AR patients; it can also improve patient’s sleep quality. This method can produce more significant efficacy than fluticasone propionate nasal spray used alone.

Animals ; Cochlea ; pathology ; ultrastructure ; Diabetes Mellitus, Experimental ; pathology ; Male ; Mitochondria ; pathology ; ultrastructure ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effect of miRNA-101 on the expression of the enhancer of zeste homolog 2 (EXH2) in human androgen-independent prostated cancer LNCaP cells.

METHODSWe divided LNCaP cells into a blank control, a negative control, and a miRNA-l01 transfection group, constructed the vector by transfecting synthetic miRNA-101 mimics into the LNCaP cells, and evaluated the efficiency of transfection by fluorescence microscopy. Then we determined the expression level of EZH2 mRNA by qRT-PCR in the three groups of cells and that of the EZH2 protein in the negative control and transfection groups by Western blot.

RESULTSGreen fluorescence signals were observed in over 70% of the LNCaP cells in the transfection group after 24 hours of transfection. At 72 hours, the expression of miRNA-101 was significantly upregulated in the transfected cells (P < 0.01), that of EZH2 mRNA was remarkably lower in the transfection group (0.01 ± 0.10) than in the blank control (0.95 ± 0.40) and negative control (0.86 ± 0.30) groups (both P < 0.01), and that of the EZH2 protein was increased in the negative control but decreased in the transfection group with the extension of culture time.

CONCLUSIONmiRNA-101, with its inhibitory effect on the expression of EZH2 in LNCaP cells, is a potential biotherapeutic for prostate cancer.

Androgens ; Cell Line, Tumor ; Enhancer of Zeste Homolog 2 Protein ; Genetic Vectors ; Humans ; Male ; MicroRNAs ; physiology ; Polycomb Repressive Complex 2 ; genetics ; metabolism ; Prostatic Neoplasms ; metabolism ; RNA, Messenger ; metabolism ; Transfection