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.

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: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.

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: 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 survey the biliary flora in patients with obstructive jaundice due to pancreatic head cancer,also the multiple factors which affect the positive findings of bile culture in these patients.Methods The information of 65 patients with obstruetive jaundice due to pancreatic head eancer,who admitted to surgery in Huashan Hospital from Oetober 2007 to October 2008 were reviewed retrospectively.The factors which may potentially affect the detection of bile pathogen in patients with malignant obstructive jaundice were studied with univarite analysis and muhivariate analysis,including age,history of biliary surgery,yellow stained time,serum alanihe aminotransferase level,serum bilirubin level,CA19-9 level,tumor size,site of obstruction,with or without clinical manifestations of biliary infection,and APACHE Ⅱ score.Results Twenty-five positive cultures happened in 65 bile samples (38.5％),including 21 strains of Gram-negative baeilli (72.4％),6 strains of Gram-positive bacteria (20.7％),and 2 strains of fungi (6.9％).Univariate analysis showed that the relevant factors which may affect the rate of positive bile culture in patients with malignant obstructive jaundice were age,history of biliary surgery,biliary obstruction site,biliary tract infection symptoms and APACHE Ⅱ score.Multivariate analysis showed that age,history of biliary surgery,biliary obstruction site and APACHE Ⅱ seore were independent risk factors.Conctusion Age,history of biliary surgery,biliary obstruction site and APACHE Ⅱ score were independent risk factors which led to positive findings of bile cultures in patients with obstructive jaundice due to pancreatic head cancer.

Objective To investigate the changes of the expression of HIF-1α (hypoxia induciblefactor-1a) mRNA in myocardium of rats after cardiopulmonary resuscitation (CPR) and the intervention effects of exogenous hydrogen sulfide on it. Method Forty five male SD rats were randomly divided into three groups: control group ( n = 15 ), cardiopulmonary resuscitation group ( CPR group, n = 15 ) and NaHS + CPR group (n = 15 ) . Rats of control group were anesthetized and intubated without asphyxia and cardiac arrest. The rats of CPR group and NaHS + CPR group were operated to induce cardiac arrest by asphyxiation. In the rats of NaHS + CPR group, NaHS in dose of 50 ug/kg was administrated via the femoral venous line 1 minute before CPR. Hemodynamic was monitored continuously. The expression of HIF - lα mRNA in myocardium of rats in each group was determined by using RT-PCR and the activity of myeloperoxidase (MPO) in the myocardium of rats in each group was assayed by using a patent reagent box 6 h after CPR. The histopathological changes of myocardium were also observed. The t- test was used for statistical analysis. Results There was no statistically significant difference in hemodynamic changes between CPR group and CPR + NaHS group ( P ＞ 0. 05 ) . When compared with the control group, the activity of MPO and the expression of HIF-1α mRNA in CPR group and CPR + NaHS group were both increased, and those increased in CPR + NaHS group was more significant (P ＜ 0. 05) . When compared with CPR group, the expression of HIF-1α mRNA in CPR + NaHS group was higher, however, the activity of MPO in CPR + NaHS group was lower ( P ＜ 0. 05) . There were various histopathological changes of myocardium of rats found in CPR group and CPR + NaHS group, and the damage of myocardium of rats in CPR group was more obvious than that in CPR + NaHS group. Conclusions The expression of HIF-1α mRNA in myocardium of rats was increased after CPR. Exogenous hydrogen sulfide can protect myocardium cells from resuscitation-perfusion injury, and the protection is associated with increase in expression of HIF 1 αmRNA.