Sleep is a vital phenomenon of life,the disturbance of which could be associated with a wide range of diseases,such as depression,anxiety,memory loss and hypertension. After years of efforts,pharmacological research of sleep in China has been in line with the pace of international sleep study,especially in the mechanisms of sleep. Our research is beginning to involved in several hot spots of study,such as the relationship between sleep disorders and their comorbidities(PTSD, depression,hypertension,diabetes and neurodegenerative diseases). Attention has also been paid to the research and discovery of novel hypnotic drugs. Despite the gap in sleep research between China and other developed countries,sleep study in China will definitely step into a gold period as well as solve sleep problems for more patients as soon as possible with the joint efforts of researchers and with the increasing attention to healthy sleep.

Objective To compare the effect of freeze-thaw tumor cells and the supernatant from tumor cell culture on the activation of lymphocytes. Methods Malignant melanoma B16 cells were prepared as tumor cell vaccine and the supernatant from tumor cell culture was collected at different time point.Transwell method was used to determine the chemotaxis of lymphocyte attracted by freeze-thaw tumor cells and the supernatant from tumnor cell culture. The cytotoxic activity of lymphocyte was detected by CCK-8 method. Results Freeze-thaw tumor cells and the supernatant from more than 2 h of tumor cell culture were found to show chemotaxis of lymphocyte. The chemotaxis of tumor cell culture more than 4 h was stronger than freeze-thaw tumor cells. Each group of chemotactic lymphocytes demonstrated to have the activity of killing tumor cells. The ability of killing tumor cells induced by the tumor cell culture more than 4 h was stronger than that induced by freeze-thaw tumor cells.In a certain range,the ability of lymphocyte chemotaxis and activation were enhanced over time. Conclusion The chemotaxis and cytotoxic activation of lymphocyte induced by the supernatant from tumor cell culture for a certain time are stronger than those by freeze-thaw tumor cells. The supernatant from tumor cell culture can be used as tumor antigen to get better immune activation instead of the freeze-thaw tumor cells.

Radical pancreaticoduodenectomy is the only effective method for the treatment of malignancies in the pancreatic head and the periampulary region.Early determination of the involvement of the superior mesenteric artery (SMA)is important for the selection of the surgical procedure and judgment of the prognosis.The operation should follow the principle of tumor-free and adequate resection range,safe resection margin and complete lymph node resection.For this purpose,we performed the radical pancreaticoduodenectomy via mesenteric approach.The SMA was dissected first,and then the tumor was en-bloc resected.From December 2011 to December 2012,24 patients with tumors in the pancreatic head or the periampullary region received radical pancreaticoduodenectomy via the mesenteric approach at the Second Affiliated Hospital of Harbin Medical University,and the short-term outcome was satisfactory.

To explore the therapeutic effect of micro-traumatic surgery of loose seton and cutting seton by rubber bands in the treatment of high anorectal fistulae.Application of cutting seton (truss rubber bands) implemented the high part of fistulae and loose seton (ligation rubber band but non-fastened) for the low part of fistulae.133/136 patients undergoing micro-traumatic surgery were cured by one operation,2 cases had pseudo-healing and there was 1 recurrent case.And the curative rate was 97.8％.The microtraumatic surgery of loose seton and cutting seton by rubber bands in the treatment of high anorectal fistula has such multiple advantages as small incision,minor trauma,lesser pain,faster healing and a shorter course of treatment.And it may preserve the proper anal function and the integrity of anal skin.And its clinical efficacy is satisfactory.

Objective To investigate the expression of long intergenic non-protein coding RNA-regulator of reprogramming (Linc-ROR) in high-grade ovarian serous cancer, and explore the relationship between Linc-ROR expression and biological function of high-grade ovarian serous cancer. Methods A total of 34 high-grade ovarian serous cancer tissue samples and 19 normal fallopian tube tissue samples were collected between June 2014 and February 2016. Real-time reverse transcription (RT)-PCR was used to detect the Linc-ROR mRNA expression in different samples. The relationship between Linc-ROR expression level and ovarian cancer International Federation of Gynecology and Obstetrics (FIGO) stage, lymph node metastasis was analyzed. Constructed Linc-ROR small interference RNA (siRNA) and pIRES2-EGFP-Linc-ROR plasmid, then Linc-ROR siRNA and pIRES2-EGFP-Linc-ROR plasmid were respectively transfected into SKOV3 cells. Cell proliferation, migration and invasion ability were assessed by cell counting kit-8 (CCK-8), wound healing assay and transwell invasion assay. Results (1) The expression level of Linc-ROR mRNA was significantly higher in high-grade ovarian serous cancer than normal fallopian tube tissues (4.31± 0.38 vs 1.03 ± 0.21; t=25.842, P<0.01). With the progression of FIGO stages, the expression of Linc-ROR was increased (F=95.702, P<0.01), and it was associated with lymph node metastasis (t=7.397, P<0.01). (2) The results of RT-PCR showed that the expression level of linc-ROR in Linc-ROR-i group was significantly lower than that in Linc-ROR-NC-i group (0.30 ± 0.11 vs 1.02 ± 0.10; t=15.269, P<0.01). The expression level in Linc-ROR-p group was significantly higher than that in Linc-ROR-NC-p group (8.90 ± 0.45 vs 1.03 ± 0.17;t=21.934, P<0.01). The CCK-8 assay showed that when the cells were cultured for 3, 4, 5 and 6 days, the A value in Linc-ROR-i group was significantly lower than that in Linc-ROR-NC-i group (P<0.05). And the A value in Linc-ROR-p group was significantly higher than that in Linc-ROR-NC-p group (P<0.05). Wound healing assay showed that, after 48 hours incubation, migration rate of cells in Linc-ROR-i group was significantly less than that in the Linc-ROR-NC-i group [(52±4)%vs(67±5)%;t=5.720,P<0.01]. The migration of cells in Linc-ROR-p group was significantly greater than that in the Linc-ROR-NC-p group [(84±4)%vs(66±4)%;t=7.330,P<0.01]. Cell transwell invasion assay showed that, after 48 hours of incubation, the number of invasive cells in Linc-ROR-i group was lower than that in Linc-ROR-NC-i group (74 ± 3 vs 104 ± 3; t=15.810,P<0.01). And the number of invasive cells in Linc-ROR-p group was higher than that in Linc-ROR-NC-p group (217 ± 4 vs 108 ± 5; t=38.060, P<0.01). Conclusion Highly expressed Linc-ROR could enhance the proliferation, migration and invasion ability of high-grade ovarian serous cancer cells, which may be one of the important molecules in the occurrence and development, invasion and metastasis of high-grade ovarian serous cancer.

Objective To investigate the mechanism of interlekin-6 (IL-6) in wound healing of human biliary epithelial cells ( BECs ).Methods BECs were cultured in IL-6 at different concentrations:0 ng/L(0 ng/L group),10 ng/L (10 ng/L group),50 ng/L (50 ng/L group),100 ng/L (100 ng/L group),1000 ng/L ( 1000 ng/L group).The effects of IL-6 on the phosphorylation of signal transducer and activator of transcription 3( STAT3 ) and the expression of trefoil family factors 3 (TFF3) were detected.BECs were divided into untreated group,STAT3-RNAi group (BECs transfected with STAT3 RNAi adenovirus) and Control-RNAi group (BECs transfected with vacant RNAi adenovirus).The effects of IL-6 on the expression of TFF3 were detected after RNAi of STAT3.In vitro wound models were constructed for the untreated group,STAT3-RNAi group and Control-RNAi group,and the effects of IL-6 and TFF3 on BECs of the 3 groups were detected.All data were analyzed by using the Student's t test,analysis of variance or Sidak test.Results The expressions of phosphorylated STAT3 in the 50 ng/L group,100 ng/L group and 1000 ng/L group were 0.240 ± 0.052,0.714 ± 0.124,0.327 ± 0.069,respectively,which were significantly higher than 0.033 ± 0.011 of the 0 ng/L group (q =5.246,17.260,7.451,P ＜ 0.05 ).The contents of mRNA and protein of TFF3 increased as the increase of IL-6 concentration (q =12.045,9.889,P ＜ 0.05).After RNAi of STAT3 of the BECs,the expression of TFF3 decreased when the concentration of IL-6 was 1000 ng/L.The expression of TFF3 of the STAT3-RNAi group was 0.037 ± 0.005,which was significantly lower than 0.267 ± 0.038 of the Control-RNAi group and 0.301 ± 0.042 in the untreated group ( q =12.135,13.929,P ＜ 0.05 ).In the in vitro wound model,the speed of BECs migration in the STAT3-RNAi group was (9.1 ± 1.5 ) μm/h,which was slower than (25.1 ± 3.8 ) μm/h of the Control-RNAi group after 12 hours of interference with IL-6 (q =7.737,P ＜ 0.05 ).The speed of BECs migration of STAT3-RNAi group was (39.2 ± 4.7) μm/h after adding 1 g/L of recombinant TFF,which was significantly faster than that of the Control-RNAi group (q =14.507,P ＜0.05).Conclusion IL-6 promotes cell migration and wound healing by activating STAT3 and up-regulating TFF3 expression.

Objective To explore the expression of frizzled-7 and β-catenin proteins in hepatocellular carcinoma (HCC),and determine their relationship with clinicopathological features and prognosis.Methods Expression levels of frizzled-7 and β-catenin proteins were detected by the SP immunohistochemical technique in 64 cases of HCC and 15 normal liver tissues.Results Frizzled-7 and β-catenin proteins were found in 42 (65.6％) and 45 (70.3％) of tumor specimens respectively,which was significantly higher than that in normal liver tissues.The expression of frizzled-7 protein was significantly positively correlated with that of β-catenin (P ＜ 0.05) in HCC.The high expression of frizzled-7 was closely correlated to tumor size (P =0.014),histologic grade (P =0.020),portal vein tumor thrombus (P =0.034),tumor recurrence (2 years,P =0.030),TNM stage (P =0.022),and HBsAg (P =0.025),and negatively correlated with 5-year postoperative survival (47.6％ vs.13.2％).The expression of β-catenin protein was significantly associated with histologic grade (P =0.012),tumor recurrence (2 years,P =0.010),and TNM stage (P =0.026),and negatively correlated with 5-year postoperative survival (36.8％ vs.20.0％).Conclusions Frizzled-7 is overexpressed in HCC and associated with decreased postoperative survival.Moreover,frizzled-7 may up-regulate the expression of β-catenin and promote β-catenin-mediated tumor invasion and recurrence.

Objective To investigate the changes of cardiac function at pre -and post -treatment in preterm infants with patent ductus arteriosus (PDA)in order to guide drug treatment.Methods Totally 84 preterm infants with PDA admitted to Neonatal Intensive Care Unit of Xuzhou Hospital Affiliated to Medical College of Southeast University from July 201 2 to June 201 4 were divided into 4 groups according to treatment drug:Ibuprofen group (27 cases),Indo-methacin group (24 cases),control group (1 1 cases),and Paracetamol group (22 cases).Patients were also divided into symptomatic PDA group (38 cases)and asymptomatic PDA group (46 cases)according to severity;PDA closed group (69 cases)and PDA unclosed group (1 5 cases)according to sequel.The level of plasma brain natriuretic pep-tide (BNP),cardiac troponin I (cTnI),correct QT intervals dispersion (QTcd)were monitored pre -and post -treat-ment.Data were analyzed by using SPSS 1 9.0 software.Results Three cardiac markers at post -treatment were of no significant difference among 4 treatment drugs.The changes of the cTnI and QTcd at pre -and post -treatment were of no significance.The level of BNP in symptomatic PDA group was significantly higher than that in asymptomatic PDA group at pre -treatment [(378 ±94)ng/L vs (1 47 ±75)ng/L,t =2.584,P =0.01 4].In the symptomatic PDA group,the level of BNP at post -treatment [(1 82 ±81 )ng/L]was significantly decreased than that at per -treatment (t =2.741 ,P =0.009).In the asymptomatic PDA group,there was no significant difference between the pre - and post -treatment [(1 21 ±61 )ng/L]in the level of BNP (t =1 .254,P =0.207).There was no significant difference in the level of BNP at per -treatment between PDA closed group and PDA unclosed group [(274 ±91 )ng/L vs (289 ± 87)ng/L,t =-0.874,P =0.391 ].In PDA closed group,the level of BNP at post -treatment [(1 21 ±74)ng/L] was significantly decreased compared with that at per -treatment (t =3.580,P =0.000).In PDA unclosed group, there was no significant difference between the pre - and post -treatment [(245 ±74)ng/L]in the level of BNP (t =0.854,P =0.392).Conclusion Early medication intervention for symptomatic PDA of preterm infants is benefi-cial for the closure of PDA and for attenuating negative effects on cardiac function of PDA.

Objective:To screen the time points of high survival rate and efferocytosis dysfunction of rat alveolar macrophages stimulated by cigarette smoke extract (CSE), establish an in vitro model of alveolar macrophage efferocytosis function, and study chronic respiratory diseases with chronic inflammatory reaction as the main pathological changes. Methods:① Time point screening experiment: rat alveolar macrophages (NR8383 cells) were cultured in vitro, and the cells in logarithmic growth phase were divided into blank control group (100 μL complete medium) and 5% CSE group (90 μL complete medium + 10 μL 100% CSE). Alma blue method was used to detect the effect of 5% CSE on the activity of NR8383 cells at 6, 12, 24 and 48 hours. ② Apoptosis induction experiment: rat type Ⅱ alveolar epithelial cells (RLE-6TN cells) were cultured in vitro as phagocytic target cells of NR8383 cells, and the cells in logarithmic growth phase were divided into blank control group and 10, 30 and 60 minutes groups after ultraviolet exposure (apoptosis was induced by 30 000 μJ/cm 2 ultraviolet irradiation for 15 minutes). Flow cytometry was used to detect the apoptosis rate of RLE-6TN cells cultured for 10, 30 and 60 minutes after ultraviolet exposure. ③ Cell efferocytosis experiment: NR8383 cells in logarithmic phase were divided into blank control group and 5% CSE group. Two hours before NR8383 cells were stimulated by CSE for 6, 12 and 24 hours, RLE-6TN cells were exposed to ultraviolet to induce apoptosis, and the RLE-6TN cell suspension was added to NR8383 cells (the ratio of RLE-6TN cells to NR8383 cells was 5∶1). Flow cytometry was used to detect the efferocytosis rate of NR8383 cells to RLE-6TN cells at different time points treated with 5% CSE. Results:① Compared with the blank control group, the activity of NR8383 cells significantly decreased after treatment with 5% CSE for 48 hours [cell reduction rate: (68.5±4.1)% vs. (73.6±2.3)%, P < 0.05]. However, there were no significant differences when the activities of NR8383 cells treated with 5% CSE for 6, 12 and 24 hours were compared with the blank control group, so these three time points were selected for the subsequent establishment of alveolar macrophage cell efferocytosis dysfunction in vitro model experiment. ② Compared with the blank control group, the apoptosis rate of RLE-6TN cells significantly increased at 10, 30 and 60 minutes after ultraviolet exposure [(66.87±8.63)%, (85.51±2.39)%, (96.13±2.74)% vs. (9.13±3.17)%, all P < 0.01] in a time-dependent manner. Considering that it taked about 50 minutes for RLE-6TN cells to be labeled with PKH26 membrane labeling probe, 10 minutes after ultraviolet exposure was selected to label RLE-6TN cells. ③ Compared with the blank control group, the efferocytosis function of NR8383 cells was significantly decreased after treatment with 5% CSE for 12 hours [cell efferocytosis rate: (33.64±1.30)% vs. (44.02±2.71)%, P < 0.01], but there was no significant effect on the efferocytosis function of NR8383 cells at 6 hours and 24 hours. Conclusions:CSE can induce alveolar macrophage cell efferocytosis dysfunction. Based on the test results of the effect of 5% CSE on NR8383 cell activity and cell efferocytosis function, 12 hours with high survival rate and weak efferocytosis effect of NR8383 cells can be selected as the in vitro model condition of alveolar macrophage cell efferocytosis dysfunction.

BACKGROUND:Previous studies have shown that home-made porous tantalum has non-toxicity and good biocompatibility, and can promote osteogenesis. Herein, we explore the mechanisms of tantalum-bone interface osseointegration.OBJECTIVE:To observe the morphological characteristics and expressions of integrin β1 and fibronectin on the interface between porous tantalum and bone tissues after implantation into the right rabbit femur, and to evaluate the biological mechanisms of tantalum-bone interface osseointegration.METHODS: Animal models of bilateral femoral condyle defects were made in Japanese big ear rabbits. Porous tantalum rod and allogeneic bone were respectively implanted into the left (experimental group) and right (control group) femur of rabbits. The animal specimens at the bone defect region were taken and made into paraffin sections and hard tissue sections at postoperative 2, 4, 8 weeks for morphological observation of new bone at the junction between the tantalum rod and host bone under light microscope, for osteogenic observation of the tantalum-bone interface under scanning electron microscope, and for immunohistochemical detection of integrin β1 and fibronectin expression.RESULTS AND CONCLUSION:Porous tantalum was bonded closely with the host bone. The loose and thick fibrous capsule was observed in the early stage and became thinner in the late stage shown by hematoxylin-eosin staining. The new bone was visible on tantalum-bone interface. Hard tissue slicing observation showed that the new bone was seen on the porous tantalum-bone interface, blood capillaries grew into the pores at postoperative 2 weeks and the pores were full of new bone tissues at postoperative 4 and 8 weeks. Under the scanning electron microscope, the osteoblasts appeared on the tantalum surface and in the pores at the early stage, and bone maturation and lamelar bone were seen at the late stage. The immunohistochemical results showed that the expression of integrin β1 in the experimental group was significantly lower than that in the control group at postoperative 2 weeks (P < 0.05), but the expression of fibronectin had no significant difference between the two groups (P > 0.05). In addition, there was a decline trend in the expression of integrin β1 and fibronectin atpostoperative 2, 4, 8 weeks. To conclude, the porous tantalum material is beneficial to enhance adhesion of osteoblasts on the surface and inside the micro-pores. Increased expression of integrin β1 and fibronectin on the tantalum-bone interface at early stage may promote early osteogenesis, while their decreased expression at bone maturing stage can promote osseointegration and bone remodeling.