Objective To identify stable reference genes for a comparison of the transcription levels of target host genes under viral infection in order to provide data for studies on interactions between the host and the influenza virus.Methods Reverse transcription quantitative real-time PCR(RT-qPCR)was performed to detect the relative expression levels of six candidate reference genes,including glyceraldehyde 3-phosphate dehydrogenase(GAPDH),β-actin,18S RNA,β2-microglobulin(B2M),ubiquitin-conjugating enzyme E2D2(UBE2D2),and ribosomal protein L37A(RPL37A)in classical cell models(A549 cells and THP-1 cells)under different conditions.The stability of the reference genes was evaluated using such methods as BestKeeper,GeNorm,NormFinder,and comparative A Ct method.Results The stability of reference genes varied depending on conditions.When such experimental factors as influenza virus infection and immune activation were taken into consideration,β-actin and GAPDH were identified as the most stable reference genes in A549 cells and THP-1 cells,followed by UBE2D2 and B2M.Conclusion The optimal reference genes in A549 cells and THP-1 cells under influenza virus infection or after being treated with interferons or LPS have been identified,which is of referential value for studying the mechanisms of viral infections.

Objective:To explore the level of tibial growth plate chondrocyte mitophagy in young rats with chronic renal failure (CRF) and its effect on chondrocyte apoptosis.Methods:Male 4-week-old Sprague-Dawley rats were randomly divided into two groups according to random number table method: normal control group ( n=20, intragastric administration with distilled water) and CRF group ( n=20, given adenine suspension 150 mg·kg -1·d -1). All the young rats were sacrificed after continuous gavage for 6 weeks. The length of tibia was measured on X ray film, the width of tibia growth plate was measured and compared on histological section, and the apoptosis rate of chondrocytes in growth plate was detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay. The growth plate chondrocytes of two groups were isolated and cultured to the third generation in vitro, and the apoptosis rate of chondrocytes was detected by TUNEL assay. The co-localization of mitochondria and autophagy lysosomes in chondrocytes was observed by double fluorescence staining. Western blotting was used to detect the level of mitochondrial marker protein translocate of the outer mitochondrial membrane-20 (Tom-20) and autophagy marker light chain-3 protein (LC-3). The mitophagy of growth plate chondrocytes was observed by transmission electron microscope. Results:Compared with the normal control group, the tibia length of CRF group was shorter [(27.32±5.81) mm vs (35.43±3.61) mm, t=5.226, P<0.001], and the relative width of growth plate in histological section was narrower (0.56±0.19 vs 1.00±0.21, t=6.744, P<0.001). The apoptosis rate of chondrocytes in growth plate in CRF group was higher than that in the normal control group (17.2%±4.8% vs 5.1%±3.4%, t=6.505, P<0.001). The apoptosis rate of chondrocytes cultured in vitro in CRF group was higher than that in the normal control group (11.8%±6.2% vs 3.1%±1.2%, t=4.357, P<0.001). The result of double influorescence staining showed that there was co-localization between mitochondria and autophagy lysosomes in CRF group. Western blotting results showed that the levels of LC-3 protein ( t=8.944, P<0.001) and Tom-20 protein ( t=6.708, P<0.001) in CRF group were lower than those in the normal control group. Conclusion:The level of tibial growth plate chondrocyte mitophagy in young rats with CRF increases, which will lead to a decrease in the number of mitochondria, an increase in the apoptosis and a decrease in the number of chondrocytes, and eventually lead to dysplasia of tibia.

Psoriatic arthritis (PsA) is a complicated psoriasis comorbidity with manifestations of psoriatic skin and arthritic joints, and tailoring specific treatment strategies for simultaneously delivering different drugs to different action sites in PsA remains challenging. We developed a need-based layered dissolving microneedle (MN) system loading immunosuppressant tacrolimus (TAC) and anti-inflammatory diclofenac (DIC) in different layers of MNs,

Objective To investigate the feasibility and effect of synchronization imaging technology of 64-slice spiral CT on urography and angiography(CTUA).Methods 36 adult patients with urinary symptoms that apply for the conventional CTU were selected. Exclusion of iodine allergy,serious heart,liver and renal insufficiency,multiple myeloma,long-term diabetes,hyperthyroidism,etc.All patients were placed intravenous tube.100 mL iohexol(300 mg I/mL),divided into 40-60 mL and 35-40 mL,shall be injected respectively by interval about 25-45 min,and the corresponding injection rate was 2-2.5 mL/s and 3.5-4.0 mL/s.About 20-30 s after the second injection,scan urinary tract for a quick CTUA source image,then scan urinary tract again for a quick source image of CTU with 5 min delay.Two groups of data with reconstruction technique(including MPR,MIP,VR etc.)should be read and diagnosed independently and double-blinded by two doctors with many years'experience in CTU.They also need to evaluate the image quality of the kidneys,urinary tracts and renal vessels,and make a statistical analysis to judge whether the CTUA inspection technique was feasible and effective.Results CTUA was good at showing renal parenchyma,ureters,urinary bladder and renal vessels,and the display rate was respectively:0.83,0.916,1,0.806,by contrast,the conventional CTU was 0,0.972,1,0,accordingly.Compared with the conventional CTU,CTUA was obviously different from the CTU,especial in displaying the distinction between renal cortex and medulla,renal vessels,and CTUA was better than the conventional CTU.While in display of ureters and urinary bladder,they had no difference,both show good.Conclusion CTUA inspection technology is superior to the conventional CTU technique,and using a routine dose contrast medium with twice bolus injection and one scanning can provide information such as the kidneys,urinary tract and renal vessels.

Objectives To investigate the clinical features of clomiphene citrate (CC) resisitant in patients with the polycystic ovary syndrome(PCOS). Methods A total of 145 women with PCOS were analyzed in the center for reproductive medicine of our hospital. According to the responses to CC ,patients were divided into two groups:(1)the CC resistant group(n = 32),(2)the CC response group(n = 113). The clinical features were compared between these two groups. Results (1)Body weight,body mass index(BMI),blood pressure and F?G scores were significantly higher in the CC resistant group than those in the CC response group (P < 0.05, respectively).(2)Basic antral follicle counts(AFC),C reaction protein(CRP),fasting insulin(FI)and HOMA?IR in the CC resistant group were significantly higher than those in the CC response group (P < 0.05, respectively),however,no significant difference was observed in sex hormones between these two groups. Conclusion PCOS patients with CC resistant are more likely to have higher body weight,BMI,blood pressure, insulin resistant?indexes and basic AFC ,the clinical doctors should consider these features before the treatment of ovulation induction.

Considering the weak relevance between the stages of C,D,I and O and the weak operability of CDIO theory in teaching,mind map is introduced into CDIO theory to drive medical English teaching.Firstly,how mind map and CDIO theory are mutually complementary is discussed,based on which,along the main clue of CDIO theory,a further discussion is carried out from four levels:conceiving the main structure and content of a passage,designing sentence teaching,implementing vocabulary memorization and operating self-directed learning,all with mind map as an assistance.Moreover,specific cases are used to exemplify how mind map is applied to CDIO medical English teaching.Using.mind maps in CDIO medical English teaching is highly coherent and operable.It helps to integrate the four stages of C,D,I and O and enhances teaching quality and learning efficiency.

The purpose of this study was to compare the pharmacokinetic profiles of tetramethylpyrazine phosphate (TMPP) in plasma and extracellular fluid of the cerebral cortex of rats via three delivery routes: intranasal (i.n.), intragastric (i.g.) and intravenous (i.v.) administration. After i.n., i.g. and i.v. administration of a single-dose at 10 mg/kg, cerebral cortex dialysates and plasma samples drawn from the carotid artery were collected at timed intervals. The concentration of TMPP in the samples was analyzed by HPLC. The area under the concentration-time curve (AUC) and the ratio of the AUCbrain to the AUCplasma (drug targeting efficiency, DTE) was calculated to evaluate the brain targeting efficiency of the drug via these different routes of administration. After i.n. administration, TMPP was rapidly absorbed to reach its peak plasma concentration within 5 min and showed a delayed uptake into cerebral cortex (t max=15 min). The ratio of the AUCbrain dialysates value between i.n. route and i.v. injection was 0.68, which was greater than that obtained after i.g. administration (0.43). The systemic bioavailability obtained with i.n. administration was greater than that obtained by the i.g. route (86.33% vs. 50.39%), whereas the DTE of the nasal route was 78.89%, close to that of oral administration (85.69%). These results indicate that TMPP is rapidly absorbed from the nasal mucosa into the systemic circulation, and then crosses the blood-brain barrier (BBB) to reach the cerebral cortex. Intranasal administration of TMPP could be a promising alternative to intravenous and oral approaches.

BACKGROUND:Study confirmed that the de-epidermized dermis(DED)can be used as dermal substitute and may form epidermal structure after incubating keratinocytes.However,the cell biological activity,tissue structure characteristics and the basement membrane component analysis of dermal substitute have been reported less.OBJECTIVE:To investigate the cell activity and the tissue structure characteristics of DED.METHODS:Skin flap was treated with 56℃ phosphate buffered solution to remove the epidermis,and the dermal cell components were deleted by freezing and thawing with liquid nitrogen to obtain DED.The DED cell activity was detected with tissue culture method,hematoxylin nuclear staining was used to determine the DED cell nuclei,and vimentin immunohistochemistry was applied for fibroblast determinations.The basement membrane and its components were detected using Periodic Acid-Schiff staining and collagen type Ⅳ immunohistochemistry.Van Gieson stain,Weigart stain and those double staining were respectively used to determine DED collagen fibers and elastic fibers.The DED ultrastructure was observed under transmission and scanning electron microscope.RESULTS AND CONCLUSlON:Using tissue culture method,the cultured DED did not exhibit cell growth at 2 weeks.Hematoxylin-eosin staining showed no nuclear in DED,vimentin immunohistochemistry showed no vimentin expressed in DED.Van Gieson staining showed DED collagen fibers were stained as rose red,Weigert staining showed DED elastic fibers were stained as pu rplish black double staining further demonstrated uniform arrangement of collagen fibers and elastic fibers.DED surface and the remaining appendages were strongly positive for Periodic Acid-Schiff staining,and type Ⅳ collagen expression was significant.Transmission and scanning electron microscope results showed that,the DED elastic fibers and collagen overlap arranged with pore intervals,they intercrossed into a network.There is no living cell component in DED,dermal matrix surface and appending organ luminal wall still retain glycogen,type Ⅳ collagen and other basement membrane components,dermal matrix is rich in collagen and elastic fibers.it is a three-dimensional collagen matrix similar to in vivo dermis.

AIM: To explore the mutual effect of co-culture of wild-type astrocytes (ASC) and motor neurons VSC4.1 (VSC) on the respective ability to produce reactive oxygen species (ROS). METHODS: The inhibition rates of cell growth in ASC and VSC in co-culture or independent culture was detected after exposed to excitatory stimulus by MTT method. Real-time observation of ROS production by ASC and VSC labeled with Hoechst 33342 was detected by confocal microscopy under the conditions of co-culture or independent culture. RESULTS: Higher concentration of glutamate induced a higher inhibition rate in mixed cell growth than that in ASC alone, while lower concentration of glutamate induced a higher inhibition rate in mixed cell growth than that in VSC only. Real-time observation by confocal microscopy showed that ROS production by VSC under the condition of co-culture, which showed a notable increase at 15 min, was significant less than that in independent culture, which peaked at 5 min and was gradually decreased. ROS production by ASC in co-culture began to increase significantly at 10 min. CONCLUSION: Compared to independent culture, ASC reduces the resting ROS production by co-cultured with VSC, while ASC prolongs the duration of ROS production by VSC after exposed to excitatory stimulus.

Objective The purpose of this study was to explore the lived experience of infertile women who terminated treatment after in vitro fertilization (IVF) failure. Methods Using a qualitative research de-sign, 12 subjects were recruited who had experienced IVF failure for many times. Data were collected through deep interviews, and analyzed using interpretive research strategies of phenomenology. Results Informed consent was obtained from each subject. There were four themes of lived experience which emerged from the data: frustration, high pressure, giving up pregnancy, and replaning the future. Conclusions Infertile women who received in vitro fertilization treatment will eventually accept the reality and mark out their future life after experiencing hustrated sense and various pressure by failure treatment.Nurses should give more loving care and persuasion during this process.