Objective To investigate the effects of KAll gene transfection on proliferation,migration,invasion and VEGF expression of pancreatic cancer MiaPaCa-2 cells under hypoxic condition,and explore possible mechanism.Methods The pcMV-KAI1 vector which contained the full length of KAI1 cDNA was transfected into pancreatic cancer cells MiaPaCa-2,and KAI1,VEGF C,VEGF A protein expressions were determined by Western blot.The proliferation of pancreatic cancer cells was evaluated by MTT method.Wound-healing assay and cell invasion assay were used to detect the migration and invasion of pancreatic cancer cells.The expression of VEGF C,VEGF A in supernatant of culture was measured by ELISA method.Results The expression of KAI1 protein in MiaPaCa-2 K transfected with KAI1 was significantly higher than that in nontransfected cells [(0.549 ± 0.021) vs 0,P＜0.05].The proliferation under hypoxic condition was not significantly different,but the migration distance was significantly shorter and the number of transmembrane cell was significantly decreased [(14.0 ± 5.8) vs (43.0 ± 14.4),P ＜ 0.05].The expression of VEGF-C in cell was significantly decreased [(0.218 ± 0.043) vs (0.745 ± 0.069),P ＜0.05],but the expression of VEGF-A was not significantly different; the expression of VEGF-C in cell culture supernatants was significantly decreased [(1236 ± 247) vs (2045 ± 221) pg/ml,P ＜ 0.01].Conclusions The migration,invasion ability of pancreatic cancer MiaPaCa-2 cells with KAll transfection under hypoxic condition is decreased,and the possible mechanism of inhibiting lymphatic metastasis is down-regulation of VEGF-C expression.

Objective To observe the inhibitory effect on lymph node metastasis of pancreatic cancer and lymphangiogenesis in mice by injection of KAll gene within xenograft tumor.Methods Pancreatic cancer cell line MiaPaCa-2 wag used to construct the nude mice models bearing tumors,then the mice were divided into normal saline group,Ad group and Ad-KAI1 group.Since the successful model construction,normal saline,Ad,Ad-KAI1 was injected every week for 3 times,respectively in the three groups,then the tumor size was documented.50 d after model construction,the tumor and enlarged lymph nodes were collected and subjected to pathological exam,and the expression of LYVE-1 and the MLVD in xenograft tumor was detected by immunohistochemistry.Results Two weeks after MiaPaCa-2 implantation,the model was 100％ successfully constructed.The growth curve of subcutaneous tumor among 3 groups was not statistically significant (P＞0.05) ; the weights of subcutaneous tumor in the 3 groups were (2514.4 ±351.3),(2466.1 ± 295.5),(2294.4±255.4) mg after 50 d,and the difference among the 3 groups was not statistically significant (P ＞0.05).Enlarged lymph nodes metastasis was observed in 8 mice (80％) in normal saline group,and 20 lymph nodes were collected,with 2.0 lymph nodes per mice; and enlarged lymph nodes metastasis was observed in 7 mice (70％) in Ad group,and 15 lymph nodes were collected,with 1.5 lymph nodes per mice; while enlarged lymph nodes metastasis was observed in 4 mice (40％) in Ad-KAI1 group,and 6 lymph nodes were collected,with 0.6 lymph nodes per mice.All the lymph nodes were confirmed to be metastasis of the primary tumor after pathologic exam.The difference of lymph nodes metastasis,number of lymph nodes metastasis per mice among the 3 groups was statistically significant (F =3.14,3.35,P ＜ 0.05).The MLVD of subcutaneous tumor among the 3 groups was (18.70 ± 5.60),(19.40 ± 4.58),(9.80 ±4.10)/400 times magnification,the MLVD of Ad-KAI1 group was significantly lower than those in normal saline group and Ad group (F10.76,11.36,P ＜ 0.05),but the difference between normal saline group and Ad group was not statistically significant.Conclusions KAI1 can inhibit the lymph node metastasis of pancreatic cancer,and the mechanism may be related with decreased lymphangiogenesis and reduced lymphatic vessel density.

BACKGROUND:Many studies concern the comparison of micro-titanium plate fixation and suture suspension fixation during cervical posterior expansive open-door laminoplasty, but the sample size of many studies has limitations. There is lack of objective evaluation on advantages and disadvantages of micro-titanium plate.
OBJECTIVE:To systemicaly evaluate the efficacy and safety of micro-titanium plate fixationversus suture suspension fixation in cervical posterior expansive open-door laminoplasty.
METHODS: English and Chinese randomized controled trials were searched by two reviewers. They retrieved the Cochrane Central Register of Controled Trials (CENTRAL), PubMed, EMbase, the ISI Web of Knowledge Database, CNKI, CMB, VIP and Wanfang database for randomized controled trials addressing micro-titanium plate fixationversus suture suspension fixation in cervical posterior expansive open-door laminoplasty published from database foundation to March 1, 2015. The references were also searched by hand. Meta-analyses were performed by using the Rev-Man 5.3 software, provided by the Cochrane Colaboration.
RESULTS AND CONCLUSION: A total of 9 studies involving 642 patients were included. The results of meta-analyses showed that: (1) safety: There were no significant differences between the two groups in operation time [SMD=-0.02, 95%CI (-0.57, 0.54),P=0.95 > 0.05], and intraoperative blood loss [SMD=0.07, 95%CI (-0.26, 0.40),P=0.69 > 0.05]. (2) Efficacy: compared with suture suspension fixation, Japanese Orthopaedic Association Scores were higher [SMD=0.26, 95%CI (0.10, 0.42),P=0.001 < 0.05], the angle of the opened laminae was bigger [SMD=0.25, 95%CI (0.02, 0.48),P=0.04 < 0.05], cervical curvature was better [SMD=0.46, 95%CI (0.27, 0.65),P < 0.000 01], and incidence of axial symptoms was lower [RR=0.40, 95%CI(0.29, 0.56),P< 0.000 01] after micro-titanium plate fixation. These findings suggest that during expansive open-door laminoplasty for treatment of cervical spondylosis, micro-titanium plate fixation and suture suspension fixation can obtain good clinical outcomes. However, Japanese Orthopaedic Association Scores were higher and the angle of the opened laminae was better in micro-titanium plate fixation than in suture suspension fixation. Micro-titanium plate fixation could effectively prevent loss of cervical curvature and reduce the incidence of axial symptoms. For the poor quality of the original studies and smal sample size, a prudent choice is suggested. More high-quality large-sample studies are needed for further verification.

In order to know that Streptococcus exist in health swine herd,2 204 cases of nose swabs were collected in swine from different areas in Dongbei three province of northeast China.Seed swab to Streptococcus culture medium,24 hours after culturing,gram staining with becteria culture,gram positive Streptococcus were identified with PCR technology,then farther identified serotype with Streptococcus suis 1,2,7,9 serotype special primers.The results showed that Streptococcus exist rate in swine herd of Heilongjiang,Jinlin,Liaoning is 29 %,27 %,34 %,respectively.PCR typing assay indicated that 155 strains of Streptococcus suis were isolated including 7 strains of 1 serotype,39 strains of 2 serotype,4 strains of 7 serotype,11 strains of 9 serotype,94 strains of other serotype in three province of northeast China.

This study aimed at exploring the mechanism of the rapid antidepressant effects of Yueju pill on depression of Parkinson's disease (DPD).The fast antidepressant effects of Yueju pill in this study was evaluated by behavior tests,such as open field test (OFT),tail suspension test (TST),forced swimming test (FST) and sucrose preference test (SPT) according to the modeling method of subacute DPD in the literature.In vitro experiment was implemented using PC12 cells.Moreover,the protective effects of Yueju pill on 1-methyl-4-phenylpyridinium (MPP+) induced neural injury with the engagement of cAMP response element binding protein (CREB) were expounded.As a result,it was found that the immobility time of the mice in the model group was significantly longer than that in the normal group in TST and FST tests (P ＜ 0.01),and the SPT ratio of mice in the model group remarkably decreased (P ＜ 0.01).In addition,the immobility time of DPD mice was shortened in the FST test after administering Yueju pill (P ＜ 0.05),while the SPT ratio was increased (P ＜ 0.01).Yueju pill took the effects on the third day after a single administration.The phosphorylation of CREB (p-CREB) in MPP+ induced PC12 cells was decreased in comparison with the model group,while the expression of p-CREB was up-regulated with the administration of Yueju pill (P ＜ 0.01).In conclusion,DPD was quickly mitigated after the treatment of Yueju pill,the activation of CREB signaling pathway and its neuroprotective effects may be the mechanism behind it.

We constructed the eukaryotic expression vector of human IL-35-IgG4 (Fc)-pOptiVEC-TOPO by gene recombination technique and expressed the fusion protein human IL-35-IgG4 (Fc) in CHO/DG44 cells. The two components of the newly discovered cytokine human IL-35, EBI3 and IL-12p35, were amplified by PCR from the cDNA library derived from the KG-I cells after LPS induction. The two PCR-amplified cDNA fragments of human IL-35 were linked by over-lapping PCR and then cloned into the IgG4 (Fc)-pOptiVEC-TOPO vector. The constructed plasmid with the recombinant cDNA IL-35-IgG4 (Fc) was verified by restriction enzyme digestion analysis, PCR and DNA sequencing. The verified plasmid with the recombinant cDNA was transfected into CHO/DG44 cells using Lipofectamine 2000. The success of the transfection was examined and confirmed by RT-PCR. After selection in alpha-MEM (-) medium, the IL-35-Ig G4 (Fc) positive CHO/DG44 clones were chosen and the media from these positive clones were collected to be used to purify the fusion protein. The positive CHO/DG44 clones were further cultured in increasing concentrations of MTX and the expression levels of the fusion protein IL-35-Ig G4 (Fc) were repetitively induced by MTX-induced gene amplification. The IL-35-IgG4 (Fc) fusion protein was purified from the media collected from the positive CHO/DG44 clones by protein G affinity chromatography and then identified by SDS-PAGE and Western blotting. The results showed that one protein band was found to match well with the predicted relative molecular mass of human IL-35-IgG4 (Fc) and this protein could specifically bind to anti-human IgG4 (Fc) monoclonal antibody. In conclusion, our study successfully established an IL-35-IgG4 (Fc) positive DG44 cell line which could stably express IL-35-IgG4 (Fc) fusion protein.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; Gene Fusion ; genetics ; Genetic Vectors ; Humans ; Immunoglobulin Fc Fragments ; biosynthesis ; genetics ; Immunoglobulin G ; biosynthesis ; genetics ; Interleukins ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

Nine cases of colorectal lipoma who were admitted from January 2005 to December 2016 were reported.The pathological diagnosis was confirmed after surgery,and all 9 cases were cured and discharged.All patients were followed up and were alive by January 2017.

OBJECTIVETo investigate the apoptosis inducing effects of ponicidin (PON) on leukemic K562 cells and its mechanisms of action.

METHODK562 cells in culture medium in vitro were given different concentrations of PON (10-50 micromol x L(-1)) for 24, 48 and 72 h. The inhibitory rate of the cells was measured by MTT assay, cell apoptotic rates were detected by flow cytometry (FCM) using Annexin V staining after K562 cells were treated with different concentrations of PON for 72 hours, and cell morphology was observed by Wright-Giemsa staining. Western blot was used to detect caspase-3 and poly(ADP-ribose) polymerase (PARP) expression, and the protein levels in mitogen-activated protein kinase signaling pathways (MAPKs, p-P38, p-ERK and p-JNK) as well as p-AKT and p-P85 in PI3K/AKT signaling pathways were also detected.

RESULTPON (over 30 micromol x L(-1)) could inhibit the growth of K562 cells in both time- and dose-dependent manner. FCM analysis revealed that apoptotic cells were gradually increased in a dose-dependent manner after treatment for 72 hours, and that marked morphological changes of cell apoptosis such as condensation of chromatin was clearly observed by Wright-Giemsa staining after treatment by 50 micromol x L(-1) PON. Western blot showed cleavage of the caspase-3 zymogen protein (32 kD), with the appearance of its 17 kD subunit, and a cleaved 89 kD fragment of 116 kD PARP was also found. Furthermore, Western blotting also showed that expression of p-AKT and p-P85 in PI3K/AKT signaling pathways was downregulated dramatically whereas the expression of p-P38 as well as p-ERK and p-JNK remained unchanged after the cells were treated by PON for 48 h.

CONCLUSIONThe results demonstrate that PON exhibits in vitro anti-leukemia effect by induction of apoptosis in K562 cells, and that PON induced apoptosis in K562 cells mainly related to activation of caspase-3 as well as inactivation of PI3K/AKT signaling pathway via down regulation of the expression of p-AKT and p-P85 protein levels. These results provide strong laboratory evidence for further anti-leukemia trials of PON.

Apoptosis ; drug effects ; Blotting, Western ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Diterpenes ; pharmacology ; Humans ; Poly(ADP-ribose) Polymerases ; metabolism ; Signal Transduction ; drug effects

Objective To analyze the application of virtual reality technology and mixed reality techniques in our hospital before and during intraoperative evaluation of complicated congenital heart diseases .Methods Methods Retrospectively ana-lyze the clinical treatment, surgical decision-making, intraoperative and early prognosis of 11 children with complicated congen-ital heart disease assessed by virtual reality technology and mixed reality techniques.The time of operation was 34-121 min, CPB time was 26-101 min, the clamping time of aorta was 18-56 min.There was no operative death.Results All 11cases were assessed by virtual reality technology and mixed reality technology before surgery .Personalized surgical strategies were made based on the evaluation results.All patients had undergone operations successfully.Compared with traditional surgical methods, fewer surgical incisions and shorter operation time were required.And it improved the surgical results.Conclusion Virtual reality technology and mixed reality technology have a great advantage in preoperative and intraoperative evaluation of complex congenital heart diseases.They can optimize surgical strategies, shorten operation time, and reduce surgical trauma. They are worthy of further promotion and application in clinical practice.

Objective To optimize conditions for the preparation of amidoximated polyacrylonitrile (PAO) membrane for U (VI) adsorption that can be easily recycled and reused. Methods Using polyacrylonitrile powder as a raw material, the PAO membrane was prepared using the nonsolvent-induced phase separation method under different conditions. The effects of the polyacrylonitrile concentration and temperature and duration of amidoximation in the preparation of casting solution as well as alkali treatment before adsorption on the U (VI) adsorption performance of PAO membrane were investigated. Results Alkali treatment significantly improved the U (VI) adsorption performance of PAO membrane, and the temperature and duration of alkali treatment affected the U (VI) adsorption capacity. The optimal alkali treatment conditions were 0.1 mol/L NaOH, 80 min, and 60℃. The polyacrylonitrile concentration and temperature and duration of amidoximation in the preparation of casting solution influenced the adsorption performance of PAO membrane. The optimal preparation conditions were 10wt% polyacrylonitrile and amidoximation at 65℃ for 18 h. Under the optimal preparation and alkali treatment conditions, the U (VI) adsorption capacity of PAO membrane reached 255 mg/g. Conclusion The PAO membrane has the advantages of simple preparation, high adsorption capacity, and easy recycling and reuse, which can be used as an ideal material for adsorbing U (VI) in wastewater.