Objective To develop a modified method of adherent culturing NSCs to replace the traditional suspension culturing method.Methods Neural stem cells(NSCs)were isolated from brain of fetal mouse at 15.5-day fetal age.In culture bottle which had been pre-coated with poly-L-ornithine hydrochloride/fibronectin(PO/FN),the cells were adherently cultured and passaged.The cell morphology was observed under inverted microscope.NSCs and their differentiated cells were identified by immunofluorescence.Results NSCs obtained in this study were successfully adherently cultured and expressed specific markers.Conclusion NSCs are successfully adherently cultured in this study.As compared with the traditional suspension culturing method,adherent culturing is simpler and has lower wastage of passage.It is easy to observe cell morphology and differentiation by this method.It is also helpful to culturing and storage of NSCs.

Objective To study the results using T-tube and retroperitoneal space drainage to manage perforating injury of the distal common bile duct(PIDC) during common bile duct(CBD) exploration.Methods We retrospectively analyzed the clinical data of 12 patients who were diagnosed to suffer from PIDC during CBD exploration from 2010 to 2012.Result All these 12 patients who received T-tube and retroperitoneal space drainage,gastrointestinal decompression,nutritional support and antibiotics recovered uneventfully.Conclusion Given that the CBD was unobstructed,T-tube and retroperitoneal space drainage was an good treatment for patients with PIDC.

Objective To assess the safety and efficacy of an olive oil-based lipid emulsion for parenteral nutrition in patients after hepatectomy.Methods Thirty-one postoperative patients with elective hepateetomy were randomized to receive isonitrogenous,isocalorie parenteral nutrition over 6 days after liver lobectomy(48-72hours)with either olive oil-based lipid emulsion(study group,n=15)or standard soybean oil emulsion(control group,n=16).The liver function and plasma protoins were assessed using peripheral venous blood collected before surgery,one day after surgery,and 7 days after surgery.The safety profiles of emulsion supports and postoperative rehabilitation were also assessed.Results The preoperative serum levels of total bilirubin,direct bilirubin,alanine amiotransferase,aspartate aminotransferase,alkaline phosphatase,total protein,albumin,and prealbumin were comparable between the two groups(all P>0.05).Although the Postoperative safety profile and liver function were not significantly different between two groups(all P>0.05),plasma total proteins,albumin,and prealbumin returned to the normal levels significantly faster in the study group than in control group[(57.57±9.84)g/L vs.(47.76±6.53)g/L,P=0.000;(31.29±3.11)g/L vs.(26.34±4.87)g/L,P=0.000;(0.188±0.059)g/L vs.(0.103±0.037)g/L,P=0.000]on the 7th Postoperative day,and the Postoperative hospital stay was also significantly shorter in the study group[(13.1±1.2)d vs.(15.2±1.1)d,P=0.041].The incidence of postoperative complications in study group and control group was 26.7%and 31.3%.respectively.Conclusions Treatment with the new olive oil-based lipid emulsion is weU tolerated in hepatectomy patients.It can speed up plasma proteins recovery and may shorten postoperative hospital stay,although it does not remarkably decrease the incidence of postoperative complications.

The initiation and operation of the tissue repair program between the broken ends after fracture is very important for fracture healing, which goes through three intertwined and gradual evolution stages: hematoma inflammatory organization stage, primitive callus formation stage and callus remodeling stage. It is completed by a variety of tissues, cells and cytokines in the bone marrow cavity. In the study of the mechanism of fracture healing, it is found that there are many signal pathways and molecules regulating bone repair, including bone formation, bone remodeling and neovascularization. At the cellular level, it regulates osteoblasts, chondrocytes, osteoclasts and endothelial cells. Hippo signaling pathway is a signal pathway that maintains the size of organs and the balance between cell proliferation and apoptosis, and also plays an important role in maintaining bone homeostasis and bone metabolism. In the process of regulating bone development and repair, it regulates the physiological activities of cells in microenvironment through protein kinase cascade reaction and transcriptional coactivator. The upstream and downstream effectors of Hippo signal pathway directly or indirectly regulate the proliferation, differentiation and apoptosis of bone metabolic cells, and the interaction with Wnt signal pathway, Notch signal pathway and other important pathways related to bone repair show that Hippo signal pathway plays an important role in the regulation of fracture healing and may become a new target to promote fracture healing. This article reviews the regulatory mechanism of Hippo signaling pathway and its regulatory role in the process of fracture healing, and looks forward to the research prospect of promoting bone healing by using it as a target.

Objective To investigate the diagnostic significance of synchronous observation on bone marrow biopsy,bone marrow smear and peripheral blood smear in stubborn hematonosis.Methods Thirty-seven patients with difficult and complicated hematologic diseases were detected by peripheral blood smear,bone marrow smear and bone marrow biopsy.Those slides and smears were observed.Results The 37 cases with blood disease was diagnosed of aplastic anemia in 3 cases,5 cases of myelodysplastic syndrome,6 cases of primary myelofibrosis,13 cases of secondary myelofibrosis,7 cases of lymphoma with bone marrow infiltration and 3 cases of metastatic carcinoma of bone marrow.In the diagnosis of aplastic anemia,myelodysplastic syndrome,primary and secondary myelofibrosis,lymphoma and metastatic carcinoma of bone marrow diagnosis,bone marrow biopsy is superior to smear.Conclusion Bone marrow biopsy,bone marrow smear and peripheral blood smear synchronous observation can effectively increase the diagnostic rate,decrease misdiagnosis rate,and correctly identify the stage of the disease.

Background and purpose:Unbalance of apoptosis regulation plays an important role in tumor initiation and progression.Survivin and caspase-3 all take part in the regulation of cell cycle and apoptosis.We investigated the role of survivin and caspase-3 in apoptosis of gastric carcinoma,as well as its relationship with the clinicopathologic features and the prognosis of gastric carcinoma by using tissue microarray.Methods:The gastric cancer tissue microarrays were composed of tissues from 100 cases of carcinoma of the stomach without a history of chemo-radiation therapy and 30 gastric tissues of control subject.The diameter of each specimen on tissue microarrays was 2.0 mm.Expressions of survivin,caspase-3 were investigated immunohistochemically on these tissue microarrays.Tumor cell apoptosis index was also examined by TUNEL method.Of the 100 cases,47 cases were followed-up from 14 months to 13 years,in which the prognosis was analyzed.Results:Two paraffin-embedded gastric carcinoma tissue microarrays were successfully constructed,including 114 and 116 tissue spots,respectively.Immunohistochemical analysis showed that survivin expression was positive in 78 cases(78%) in this series.In contrast,no expression of survivin in control tissues was detected(P

The evolution of hepatocellular carcinoma (HCC) is a compound process which involves many kinds of genes and transductional pathways. The expression of the peptidyl-proplyl-isomerase PIN1 gene, the mutation in exon 3 of beta-catenin and its correspondent abnormal expression and their roles in the hepatocellular carcinogeneisis were investigated. Among 29 pair cases of HCC and non-carcinoma tissues, the expression of PIN1 gene was detected by immunochemical staining. Mutations in exon 3 of beta-catenin gene and differential expression of beta-catenin gene were investigated by the methods of PCR-SSCP, direct sequencing and immunohistochemical technique as well. The results indicated: (1) 44.8% (13/29) cases of HCC presented higher level of PIN1 gene expression than non-cancerous tissues (chi2=32.63, P<0.05), especially in cytoplasm and nucleus, while there was lower level of PIN1 expression in non-cancerous tissues; (2) 58.6% (17/29) HCC tissues showed beta-catenin protein accumulation in cytoplasm and nucleus. 46.2% (6/13) HCC tissues indicated beta-catenin protein accumulation with higher level of PIN1 expression, while 53.8% (7/13) HCC tissues indicated beta-catenin protein accumulation with lower level or trace of PIN1 expression (chi2=0.00, P>0.05); (3) 24.1% (7/29) of primary tumor lesions carried gene mutations in exon 3 of beta-catenin, and accompanied by beta-catenin protein accumulation. There was no mutation in non-cancerous tissues. All the mutation presented in tissues with low level of PIN1 expression. There was no mutation of beta-catenin gene in tissues with high PIN1 expression level (chi2=58.12, P<0.05). So it was postulated that the increase of PIN1 gene expression could promote hepatocellular carcinogenesis via a way different from beta-catenin gene mutation.

ObjectiveTo investigate the clinicopathologic significance of a disintegrin and metalloproteinase 17(ADAM 17) mRNA in esophageal squamous cell carcinoma (ESCC),and to evaluate its relation to clinicopathological features. MethodsThe expression of ADAM17 mRNA in 50 ESCC and 50 normal esophageal tissues were detected by RT-PCR. ResultsThe mRNA expression of ADAM17 in 50 ESCC and 50 normal esophageal tissues were 0.937±0.241 and 0.225±0.077,respectively.The expression of ADAM17 mRNA in the ESCC was much higher than in normal esophageal tissues (t=-19.899,P＜0.01).The expression of ADAM 17 mRNA was positively correlated with lymph node metastasis(t=-4.703,P＜0.01 ) and TNM staging (t=-2.652,P＜0.05).There were no correlation between the expression of ADAM17 mRNA and sex,age and histological grade (t=0.299,-0.907,-3.163,all P＞0.05).ConclusionADAM17 mRNA was higherly express in ESCC than in normal esophageal tissues and may play an important role in the development,invasion and metastasis of ESCC.It may be used as a prognostic factor.

ObjectiveTo evaluate the role of galactomannan (GM) detection in bronchoalveolar lavage fluid (BALF) in the diagnosis of invasive pulmonary aspergillosis (IPA) in elderly patients with lung diseases.MethodsThe elderly patients with lung diseases and suspected of having IPA were enrolled. BALF for culture and GM detection, and serum samples for GM detection were obtained from all participants.According to European Organization for Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG) with host factors, clinical criteria, mycological evidence and histological or cytopathological evidence, all patients were categorized into the proven, probable,possible and no IPA patients. The IPA patients were considered as IPA group while no IPA was considered as control group. ResultsAmong 76 enrolled elderly patients, 18 cases were diagnosed with IPA (1 proven, 11 probable and 6 possible), the remained 58 patients were as control group.Based on a GM index cutoff value of ≥0. 5, the sensitivity, specificity, positive predictive value and negative predictive value for BALF were 83.3％, 82. 8％, 60.0％ and 94.1％, respectively; these variables in serum were 55.60％ , 91.4 ％, 66.7 ％ and 86.9％ , respectively. Using a GM index cutoff value of ≥1.0, the specificity and positive predictive value of both BALF GM and serum GM reached 100％, but the sensitivity and negative predictive value of BALF GM were higher than serum GM (66.7％ vs. 22.2％, 90.60％ vs. 80.60％, respectively). Receiver operating characteristic curve analysis yielded a sensitivity of 83.4％ and a specificity of 96.6％ at a BALF GM index cutoff value of ≥0.725. ConclusionsGM detection in BALF seems to be a useful tool in the diagnosis of IPA in elderly patients with lung diseases,and could be widely used in future.

Objective To establish a canine model with pulmonary thromboembolism(PTE) of selective lobar pulmonary artery embolization mimicking chronic thromboembolism, to assess the effects of an inhaled nitric oxide (NO) 20 ppm on vital signs, blood gas, hemodynamic parameters and neutrophils in the alveolar of the canine model. Methods Twenty canines were divided into four groups: group 1: sham group (n=5); group 2; ischemic lung group (PTE group without embolectomy, n=5); group 3; reperfusion lung group (PTE group with embolectomy, n=5); group 4: reperfusion lung group with inhaled NO (PTE group inhaled 20 ppm NO after embolectomy, n=5). And central venous pressure (CVP), mean pulmonary arterial pressure (MPAP), pulmonary arterial wedge pressure (PAWP) and carbon monoxide (CO) were recorded, pulmonary vascular resistance (PVR) was also calculated. Vital signs, blood gases were measured before embolectomy and at 2, 4, 6 hours after the operation. Albumin in bronchoalveolar lavage fluid (BALF) was measured by chromatometry of Coomassie brilliant blue. Lung wet to dry weight ratio(W/D) was also measured. Lung tissue pathology and alveolar PMN in the left lower lobar were observed by optical microscopy. Results MPAP increased significantly at 2 hours after reperfusion [(3. 20±0.53)kPa vs. (2. 27±0. 67)kPa,F=63,P=0.02]; At 6 hours after reperfusion as compared with baseline,HR increased significantly [(175±8) beats/min vs. (155±5) beats/min, F=38.72, P=0.01],PaO_2/FiO_2 also decreased significantly (41.70±8.04 vs. 54.71±3.78,F=48.36,P=0.03). MPAP decreased significantly in group 4 as compared with group 3 at 2 hours after reperfusion [(2.53±0.4)kPa vs. (3. 20±0. 53)kPa,F=55,P=0.04]. At 4 hours after reperfusion,PaO_2/FiO_2 raised in group 4,but there was no significant difference as compared with group 3 (49.17±7.37 vs. 39.71±7.31, F=2.36, P=0. 11). The quantities of alveolar PMN infiltration in group 4 decreased significantly as compared with group 3 (19±6/10 HPF vs. 31±11/10 HPF, F=98, P=0.01).Conclusions Lung ischemia-reperfusion injury can be induced by embolectomy from lower pulmonary artery in the PTE model embolized for one week. An inhaled NO 20 ppm can decrease the elevated pulmonary artery pressure induced by ischemia-reperfusion injury and may alleviate the injury by reducing the PMN immigration into the alveoli.