Objective:To investigate the differences of gene expression and signal transduction pathways in large conductance calcium-activated potassium channels(BKCa) gene knockout rats and analyze the role of BKCa gene in pancreas.Methods:Three adult female BKCa knockout SD rats (BKCa knockout group) were donated by Professor Wang Wei from Department of Pathology and Physiology of Basic Medical College of Capital Medical University, and three wild type adult femal SD rats were used as wide-type group. The whole pancreas was resected and RNA was extracted. RNA transcriptome sequencing (RNA-seq) technology was used for sequencing and DESeq2 differentiation analysis software was used for screening differentially expressed genes between two groups, and the gene ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis were performed. The key genes were validated by RT-PCR.Results:18 258 genes were detected by sequencing in the 2 groups. There were statistically significant differences in the expression of 348 genes screened by DESeq2, 200 of which were highly expressed in the pancreas of BKCa knockout group, and 148 of which were low-expressed. 214 differentially expressed genes enrichments were found in GO database, including 25 involved in biological process, 18 in cell components and 14 molecular functions. All 348 differentially expressed genes were found in KEGG database, 15 of which were significantly enriched in PI3K/Akt signaling pathways. RT-PCR results showed that the expression of key genes Hsp90ab1, Hsp90aa1, Foxo3a and Col1a2 in the BKCa knockout group was significantly higher than that in wide type group ( P<0.0001), while Thbs1, Pik3r1 and Ppp genes were not significantly different. Conclusions:Differentially expressed genes and related important regulatory signaling pathways were screened out between BKCa knockout SD rats and wild-type SD rats at the transcriptional level, and PI3K/Akt pathway was found to be the most enriched, providing an important clue for predicting the function of BKCa in the pancreas.

OBJECTIVE:To study the effects of Shenfu qiangxin(SFQX)pills on the expression of autophagy-associated pro-tein LC3b and pro apoptotic gene Bax in myocardial cells of rats with cardiorenal syndrome (CRS). METHODS:Rats were ran-domly divided into sham operation group(water),model group(water),positive control group(Captopril tablets 2.3 mg/kg)and SFQX pills high-dose,medium-dose and low-dose groups [13.2,6.6,3.3 g(crude drug)/kg],with 10 rats in each group. CRS mod-el was induced in those groups by abdominal-aortae-constriction+acute renal ischemia reperfusion injury except for sham operation group;and they were given relevant medicine intragastrically 8 week after operation,once a day,for consecutive 4 weeks. Plasma contents of Cr and ALD,the protein expression of LC3b and Bax in myocardial tissue of rats were detected 24 h after last medica-tion;ventricular index was calculated,and morphological change of myocardial tissue was observed. RESULTS:Compared with sham operation group,the plasma contents of Cr and ALD,ventricular index and the protein expression of LC3b in myocardial tis-sue increased significantly in model group (P<0.05 or P<0.01);and myocardial cell suffered from endochylema red deletion, myocardial cross striation disorder,intercellular space fibrosis aggravation and so on. Compared with model group,the plasma con-tents of Cr and ALD(except for positive control group)and the protein expression of LC3b and Bax in myocardial tissue decreased significantly in positive control group and SFQX pills high-dose group(P<0.05 or P<0.01);myocardial pathological change was improved;the ventricular index decreased significantly in SFQX pills low-dose and medium-dose groups (P<0.05). CONCLU-SIONS:SFQX pills can decrease the plasma contents of Cr and ALD,inhibit myocardial cell autophagy and apoptosis in CRS rats.

BACKGROUND:Subcutaneous fat of human body is a rich reservoir of adipose derived stromal stem cells(ADSCs).ADSCs can proliferate rapidly when being cultured in vitro,and has the capacity of multi-directional differentiation.ADSCs attracted much attention in research of tissue engineered seed cells.OBJECTIVE:To isolate and culture stromal vascular fraction(SVF)cells from rabbit subcutaneous fat in vitro,and to testify whether it has multiple differentiation capacity.METHODS:SVF cells were isolated in vitro from rabbits,and cultured under standard condition.Cellular surface antigens CD44,CD45 and CD29 of passage 3 SVF cells were examined using flow cytometry.Passage 3 SVF calls were induced to differentiate into osteoblasts,chondrocytes,and lipocytes.Oil red staining was used to examine lipocyte induction.Alkaline phosphatase(ALP)staining,alizarin red staining and von Kossa staining were used to examine osteoblast induction.Type Ⅱ collagen immunohistochemical staining and type Ⅱ collagen mRNA RT-PCR were used to examine chondrocyte differentiation.RESULTS AND CONCLUSION:Primary SVF cells were multi-angular or short spindle-shaped.Passage 3 SVF calls were long spindle-shaped.Flow cytometry showed CD44+,CD29+,CD45.Oil red staining exhibited positive reaction in lipocyte induction group.ALP staining,alizarin red staining and Von kossa staining demonstrated positive reactions in osteoblast induction group.Type Ⅱ Collagen immunohistochemical staining and alcian blue staining have suggested positive reactions at 14 days of chondrogenic induction group.RT-PCR of type Ⅱ collagen mRNA test showed that the product band had strong signal at 14 days of chondrogenic induction group compared with that before induction.Above mentioned results have indicated that SVF cells isolated from rabbit subcutaneous fat have identical surface makers of stem cells,and have the ability to differentiate into lipocytes,ostsoblasts and chondrocytes in vitro by induction,and it could be concluded that the SVF cells were ADSCs.

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Silicosis ; diagnostic imaging ; Tomography, Spiral Computed

BACKGROUND:Conventional therapies for lumbar spondylolisthesis can result in trauma,bleeding and low back pain.With the vigorous development of spinal biomechanics and novel spinal fixation systems,we have more understanding on the reduction and fusion after spondylolisthesis.OBJECTIVE:To observe the clinical effects of transforaminal lumbar interbody fusionvia the quadrant system on lumbar spondylolisthesis and related biomechanical changes.METHODS:A retrospective analysis was done in 23 patients with lumbar spondylolisthesis undergoing transforaminal lumbar interbody fusionviathe quadrant system admitted from June 2012 to September 2013.Oswestry disability index and visual analog scale score were detected at 3 months and 1 year after treatment,as wel as fusion conditions and internal fixation with or without loosening or breakage.RESULTS AND CONCLUSION:Al patients were successfuly treated,with no cerebrospinal fluid leakage and nerve injury.Incisions were healed wel in al cases except one case suffered from incision infection that wascontroled after 10 days of antibiotic treatment.Al the patients were folowed up.The Oswestry disability indexes and visual analog scale scores were significantly improved at 3 months and 1 year after treatment (P ＜0.05),but there was no difference in these two scores at 3 months and 1 year after treatment (P＞0.05).The improvement rates of Oswestry disability index and visual analog scale score were (65.3±14.8)％and (58.2±12.0)％,respectively.These findings indicate that the transforaminal lumbar interbody fusionvia the quadrant system is safe and effective to correct lumbar spondylolisthesis,maintains the biomechanical stability,improves patient's symptoms,reduces the incidence of low back pain and improves the quality of life.

BACKGROUND: Studies demonstrated that small intestinal submucosa (SIS) had no immunogenicity, which can not lead to rejection following transplantation, thus, this is an ideal skin substitutes for natural skin.OBJECTIVE: Basic fibroblast growth factor (bFGF) gene was transfected into bone marrow mesenchymal stem cells (BMSCs),and combined the cells with SIS to construct tissue-engineered skin.METHODS: BMSCs were obtained from Japanese big-earad rabbits, and in vitro cultured. Then the subculturad BMSCs were transfected by pCDNA3.1 plasmid, followed by incubation on swine SIS to construct the tissue-engineered skin. The growth of cells and phenotype of BMSCs were detected by flow cytometry. In addition, the result of transfecting BMSCs with pCDNA-bFGF vector was measured by Western blot, and the structure of tissue-engineered skin was observed.RESULTS AND CONCLUSION: After passaged, BMSCs were grown quickly with Iong-fusiform shape. The cells were positive expressed CD90 and CD44, but negative expressed CD45. bFGF had been transfected into BMSCs, and stable expressed. The transfected BMSCs grew well in SIS. By this method, tissue-engineered skin can be constructed in vitro.

Objective To compare the curative effect of anatomical plate and locking plate in treatment of calcaneous intraarticular fracture. Methods 67 petiants with calcaneous intraarticular fracture were randomly divided into anatomical plate group (n=33) and locking plate group (n=34) . The Bo..hler angle, the Gissane angle, the length of calcaneal axis, the width and height of calcaneous and the Maryland grade were compared at 1 week and 6 month after operation. Results (1) week after operation, the Bo..hler angle, the Gissane angle, the height and width of calcaneous, the length of calcaneal axis, the Maryland grade had no significant difference between 2 groups . 6 months after operation, the Bo..hler angle, the Gissane angle, the height of calcaneous had significant differe nce between 2 groups. There was no significant difference in the length of calcaneal axis and the grade of Maryland between 2 groups. Conclusions The locking plate group is better than anatomical plate group in major anatomical measure indicators in 6 months follow up. The therapy of locking plate is worth of clinical promotion.

This study was aimed to compare the activation of daidzein and puerarin on antioxidation, NO and NOS suppression in vitro . The RAW264 . 7 cell line was used to prepare radical reaction model induced by LPS (1 μg/mL). MTT method was adopted to detect cytotoxicity of daidzein and puerarin. The DCFH-DA probe and confocal microscopy were used to examine the antioxidant ability of daidzein and puerarin. Griess reagent was adopted to test the NO level in the culture medium. And chemical colorimetry was used to detect the content in RAW264.6 cells. The results showed that daidzein and puerarin can significantly suppress the NO and T-NOS expression in RAW264.7 cells induced by LPS. It was concluded that there was no difference on the activation of antioxidant free radical between daidzein and puerarin . In this regard , daidzein can be the used as substitutes of puerarin .

Objective To validate the feasibility of the simulationmethod and the reliability of thesimulationresult through comparison between simulation and measurement of the energy spectrum from medical diagnostic X-ray (RQR-Radiation qualities in radiation beans emerging from the X-ray source assembly).Methods A simplified model of the medical diagnostic X-ray RQR radiation quality was established using code of BEAMnrc.The energy spectrum of the same RQR radiation quality were measured through a plane high-purity germanium spectrometer,and compared with the simulationresult.Results The difference of spectral distribution between measurement and simulation was less than 3％,in spite of the convolution processing not happened to the pulse height distribution measured by the spectrometer.And the spectral distribution,fluence,energy fluence,means energy distribution of the radiation was obtained using the code of BEAMDP.Conclusions As indicated above,it is possible to use the simulation of the energy distribution as a foundation for the establishment of X-ray RQR radiation quality.

To investigate the interaction and involvement of sodium hydrosulfide (NaHS), a H(2)S donor, on hippocampus of rats suffering from sepsis-associated encephalopathy, rats were subjected to cecal ligation and puncture (CLP)-induced sepsis. Adult male Sprague-Dawley rats were randomly divided into four groups: Sham group, CLP group, CLP+NaHS group and CLP+aminooxyacetic acid (AOAA, an inhibitor of H(2)S formation) group. The four groups were observed at 3, 6, 9, 12 h after treatment. We examined hippocampal H(2)S synthesis and the expression of cystathionine-β-synthetase (CBS), a major enzyme involved in the H(2)S synthesis in hippocampus. CBS expression was detected by reverse transcription polymerase chain reaction (RT-PCR). The concentrations of inflammatory cytokines (TNF-α, IL-1β) were determined in hippocampus by using enzyme-linked immunosorbent assay (ELISA). Neuronal damage was studied by histological examination of hippocampus. In CLP group, H(2)S synthesis was significantly increased in hippocampus compared with sham group and it peaked 3 h after CLP (P<0.05). Sepsis also resulted in a significantly upregulated CBS mRNA in hippocampus. The levels of TNF-α and IL-1β in the hippocampus were substantially elevated at each time point of measurement (P<0.05), and they also reached a peak value at about 3 h. Administration of NaHS significantly aggravated sepsis-associated hippocampus inflammation, as evidenced by TNF-α and IL-1β activity and histological changes in hippocampus. In septic rats pretreated with AOAA, sepsis-associated hippocampus inflammation was reduced. It is concluded that the rats subjected to sepsis may suffer from brain injury and elevated pro-inflammatory cytokines are responsible for the process. Furthermore, administration of H(2)S can increase injurious effects and treatment with AOAA can protect the brain from injury.