Objective To analyze the protein expression in the rat hippocampus by the proteomic approach.Methods Proteins from hippocampal tissue homogenates of the rat were separated by two-dimensional gel electrophoresis(2-DE),and stained with colloidal Coomassie blue to produce a high-resolution map of the rat hippocampus proteome.Selected proteins from this map were digested with trypsin,and the resulting tryptic peptides were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS).The mass spectrometric data were used to identify the proteins through searches of the NCBI protein sequence database.Results 37 prominent proteins with various functional characteristics were identified.The identified brain protein classes covered metabolism enzymes,cytoskeleton proteins,heat shock proteins,antioxidant proteins,signalling proteins,proteasome-related proteins,neuron-specific proteins and glial-associated proteins.Furthermore,3 hypothetical proteins,unknown proteins so far only proposed from their nucleic acid structure,were identified.Conclusion This study provides the first unbiased characterization of proteins of the rat hippocampus and will be used for future studies of differential protein expression in rat models of neurological disorders.

Objective To detect mite allergens in the dust of air conditioner filter-net and floating air in room. Methods Samples were collected from rooms of asthma patient and normal families with or without air conditioner. Der p 1,Der f 1 and Der 2 were determined by two monoclonal antibody-based ELISA. Results In asthma patient families,the concentration of airborne Der p 1,Der f 1 and Der 2 was (0.23 ? 0.13),(2.62 ? 1.08),(0.93 ? 0.41) ng/m3,and (0.56 ? 0.25),(4.74 ? 1.22),(2.33 ? 0.64) ng/m3 respectively before and after the air conditioner switched on,all showing a significant difference (P

Objective To study the expression of apolipoprotein H (ApoH) in childhood primary nephrotic syndrome (PNS) and to discuss its role in PNS. Methods Immunohistochemistry staining and real-time quantitative polymerase chain reaction(RT-PCR) were performed to evaluate the expression of ApoH in renal tissues of 78 patients with PNS and 14 cases of normal controls. Serum albumin, serum lipid, proteinuria and urinary retinol binding protein (RBP) were tested before renal biopsy in all patients. Tubulointerstitial lesions were investigated. Results (l)There was positive expression of ApoH in renal tissues of PNS patients and normal controls,mainly in the proximal tubules by immunohistochemistry staining. ApoH mRNA was also shown in all renal tissues by RT-PCR. ApoH protein expression was positively correlaed with its mRNA expression(r=0.264, P 0.05) whereas these data displayed no significant difference between two groups. Above expression in mesangial proliferative glomerulonephritis (MsPGN) and focal segmental glomersclerosis (FSGS) down-regulated significantly (3.30?0.28,2.82?0.36, and 10.13?3.09,10.12?1.02, respectively), as compared to those in MCNS,MN and the controls, P

The study aims at cloning the CDS fragment of erk2 gene cDNA in Inner Mongolia Cashmere Goat and analyzing its tissue-specific expression, erk2 gene cDNA was cloned by RT-PCR. The nucleotide sequence was analyzed by Blast and amino acid sequence was analyzed by online softwares SMART and Psite. The tissue-specific expression pattern of erk2 was analyzed by quantitative RT-PCR. The expression of erk2 in testis of goat was detected by Immunohistochemistry. The cloned erk2 gene cDNA (GenBank Accession No. JX569765) was 1 083 bp in length, including a complete ORF encoding 360 amino acids residues. The amino acid sequence shares 100% identity with the Bos Taurus ERK2 (Bos Taurus BC133588.1). Analysis by SMART suggests that the encoded protein contained a "TEY" structure and an S-TKc domain possessing serine/threonine kinase catalytic activity. Analysis with Psite indicates one cAMP-/cGMP-dependent protein kinase phosphorylation site, 3 protein kinase C phosphorylation sites, 5 casein kinase II phosphorylation sites, 2 protein kinases ATP-binding region signatures and one serine/threonine protein kinases active-site signature in this protein. Analysis by Psort (k-NN prediction) suggestes that this protein most probably is localized in cytoplasm. The results of quantitative RT-PCR show that the expression of erk2 mRNA was higher in heart, skin and breast, whereas lower in spleen and kidney. ERK2 protein was detected in testis by immunohistochemistry.
Amino Acid Sequence ; Animals ; China ; Cloning, Molecular ; DNA, Complementary ; genetics ; Goats ; genetics ; Male ; Mitogen-Activated Protein Kinase 1 ; genetics ; metabolism ; Molecular Sequence Data ; Real-Time Polymerase Chain Reaction ; Testis ; metabolism

Objective To develop a novel method for treating complete heart block by autotransplantation of simus node node cells to right ventricular anterior wall.Methods Twenty healthy mongrel dogs were involved in the present study.The dogs were randomly assigned to transplant group or control group(n=10).The sinus node (SN)was harvested and isolated in vitro after an electronic pacemaker was implanted and complete heart bolck was introduced.The SN cells from dogs of transplant group were injected to autogenic right ventricular wall.Commensurable culture medium was implanted to ghe same position of dogs in control group.Two weeks later,detailed electropohysiological study was performed.For investigating the variation of the rhythm,epinephrine was administrated through femoral vein to dogs of transplant group.Results Most of isolated SN cells from dogs were thin-spindle shape,and cell activity was fine.The SNs by VG stained displayed typical structural feature.2 weeks after cell autotransplantation,higher heart rates were achieved from transplant group than that in control group(P＜0.05).This rhythm was stable in 4 weeks and became faster remarkably after administration of eninephrine(P＜0.05).Conclusion SN cell of dogs tutorgrfted into right ventricular anterior wall can form new pacemaker site in ventrcle and improve ventricular rate of complete heart bolck.This pacemaker site can also be regulated by epinephrine.

Objective:To understand the clinical characteristics and prognosis of Langerhans cell histiocytosis (LCH) with skin-limited lesion.Methods:A retrospective analysis was performed on clinical characteristics and prognosis of 16 skin-limited LCH patients, out of 578 LCH patients who were hospitalized in Beijing Children′s Hospital during December 2013 to June 2018.Results:A total of 16 skin-limited LCH cases, accounted for 2.7% of all 578 cases, were included.Among which, sex ratio (male vs.female) was 1.28∶1.00.Median ages of skin eruption occurrence and of diagnosis of the disease were 3.5 months (3 days to 2 years and 5 months) and 6 months (2 months 14 days to 2 years and 8 months) in this group.Among the 16 cases, seborrheic dermatitis-like lesions(11 cases, 68.7%) was the most common, and the trunk was most frequently involved[75.0% (12 cases)]. Serine/threonine protein kinase gene V600E [ BRAF (p.V600E)] mutation was detected in pathological specimens from 10 skin-limi-ted cases, with 9 cases being positive.Plasma samples from 5 positive cases were further detected for BRAF (p.V600E) mutation, and 4 positive results were gained.Of all 16 patients, 11 cases (68.7%) were treated.Remission were achieved in 3-6 months from treatment start in patients treated whether according to the Histiocyte Society′s LCH-2009 protocol for 25 weeks(6 cases, 37.5%), or with topical mometasonefuroate for 3 months (3 cases, 18.8%). Two patients(12.5%) with solitary cutaneous lesions underwent excision biopsy (one face and one prepuce) and were considered to be in remission immediately after surgery.None of these patients suffered from the recurrence of the disease.The remaining 5 patients (31.3%) with skin-limited LCH were just evaluated regularly, and achieved remission in 3-6 months of commencing observation.Among these untreated patients, 1 with consistently positive BRAF (p.V600E) mutation in plasma had bone involvement in the 24 th month of assessment, and was then treated based on the Histiocyte Society′s LCH-2009 Protocol.No clinical or imageological evidence supporting disease progression was found on this patient.Median follow-up period was 32.8 months (2.9-63.9 months). Except one patient, none of the rest cases had active disease till follow-up ended.Two-year event free survival(EFS) of this research was (92.3± 7.4)%.There was no significant difference between EFS of treated group and that of observation group( χ2=1.250, P=0.264). Conclusions:Skin-limited LCH often occurs in infants and newborns, with strong heterogeneity in clinical manifestations, laboratory indicators, and pathogenesis.Seborrheic dermatitis-like lesions were the most common cutaneous type.The prognosis of the patients is excellent despite progressing into multisystem involvement can be seen in a few patients.

Objective To explore the feasibility of repairing porcine bile duct defect with decellularized rabbit abdominal aorta matrix scaffold.Methods Sodium dodecyl sulfate and Sodium deoxycholate were used to remove the cells in the blood vessel,and the residual DNA and RNA fragments were removed by nuclease.The prepared scaffold was implanted to repair defect of bile duct in swine,which were sacrificed after 45 days of surgery for histological evaluation.Results HE,Masson and elastic fiber staining showed that the composition and structure of the scaffold maintained their native features after dcellularization treatment.DNA content in acelllular scaffold (0.12 ± 0.01) μg/mg dry weight) was significantly decreased as compared with the native ones (2.31 ± 0.03) μg/mg dry weight,P ＜ 0.05).Collagen content was increased from (152 ±22) μg/mg dry weight in intact aorta to (177 ±21) μg/mg dry weight.Adipose derived mesenchymal stem cells with typical morphology survived well in the decellularized vascular matrix.It was observed that seeded ASCs penetrated into the inner wall of the scaffold.After transplantation,there was no leakage in the anastomosis and collapse of acellular blood vessel matrix.After 45 days of transplantation,repaired bile duct was harvested for histological evaluation.HE and Masson staining revealed that there were a large number of cells distributed in the inner wall of the scaffold,and some suspected epithelial cells and glands were found.Conclusion Decellularized aorta matrix scaffold hold great potential in serving as scaffold repairing defect of bile duct.

Objective By analyzing the problems existed in the Investigator Initiated Trial (IIT),this article put forward the corresponding countermeasures and therefore provides reference for the standardization of clinical research project management.Methods Four types of problems identified in the supervision of hospital IIT projects are analyzed according to literature review,data analysis of clinical research project,comparative study and summary.Identified problems are existed in the following aspects:scientific research supervision function,research method guidelines,technical specification of the diagnosis and treatment,scientific research design and project approval review,research funds,medical ethics committee,construction of Biobank and Regulation Conflicts.Results This article put forward 6 countermeasures for improvement:establishing and perfect IIT project scientific research supervision entity,bring in the IIT project steering group to strengthen the scientific review;Strengthen risk management to ensure medical safety,carry out IIT training,establish IIT management database information system,build a comprehensive integrated development multi-point application model of hospital BioBank.Conclusions The establishment of the hospital's IIT scientific research supervision system,management mode and technical standard system is of great importance to standardize clinical research,ensure research quality and guide the clinical research work of the hospital effectively.

Objective:To investigate the effect and mechanism of miR-195 regulating FOXK1 gene and PI3K/Akt pathway on stomach adenocarcinoma proliferation, invasion and migration ability.Methods:Public database samples were employed to analyze the expression differences and prognostic significance of miR-195 in stomach adenocarcinoma. After overexpression of mir-195-5p in two cell lines, MGC803 and AGS, altered cell proliferation, invasion, and migration abilities were detected by Alamar Blue, Wound healing, and Transwell assays. The potential target genes and binding sites of miR-195 were predicted by the starBase. Western blot was used to detect the expression levels of foxk1 and phosphorylation sites in the PI3K/Akt pathway of target genes after overexpression of mir-195-5p. A Dual-luciferase reporter assay was used to verify the relationship between mir-195-5p and foxk1. Statistical analyses were performed with IBM SPSS 22 software and R 4.0.3.Results:Our results showed a significant over-expression of miR-195 in the tumor tissues, compared with the paired normal tissues ( P<0.001) , which could inhibit the proliferation and invasion of stomach carcinoma cells and significantly correlated with survival ( P=0.011) . Moreover, our study indicated that miR-195 depressed the expression of FOXK1 and significantly reduced the activation of the PI3K/Akt pathway, which had a negative effect on the proliferation and invasion of stomach carcinoma cells. The phosphorylated Akt (s473 site) expression in the PI3K/Akt pathway was significantly decreased after overexpression of miR-195. Conclusion:Overall, our studies clarify the important function of the miR-195 in the diagnosis and therapy of patients with stomach carcinoma and reveal the FOXK1 and PI3K/Akt pathway regulation by the miR-195, which are of important clinical significance in the differential diagnosis.

OBJECTIVE To evaluate the survival and functional outcomes of advanced laryngeal squamous cell carcinoma after modified supracricoid partial laryngectomy(SCPL).METHODS We selected 65 laryngeal cancer patients treated with modified supracricoid partial laryngectomy from 2002 to 2015 in our hospital.Among them,62 cases were males,3 cases were females with an age ranged from 35 to 80 years(median age 58 years)There were 26 cases with T2 stage,32 cases with T3 stage and 7 cases with T4 stage.We selected 120 laryngeal cancer patients treated with vertical partial laryngectomy at the same period as control.Then we evaluated the functional outcomes of modified supracricoid partial laryngectomy group compared with vertical partial hemilaryngectomy group.RESULTS The 5-year cumulative survival rate and decannulation rate were 82.3％ and 98.3％ for supracricoid laryngectomy group respectively.Decannulation ratewas 86.1％ for vertical partial hemilaryngectomy group(P＜0.05).However,there was no significant difference between supracricoid partial laryngectomy group and vertical partial laryngectomy group in pronunciation evaluation and abnormal deglutition.CONCLUSION Modified supracricoid partial laryngectomy is a good choice for local advanced laryngeal squamous cell carcinoma.