Objective To study the role of NF-?B pathway in HGF (hepatocyte growth factor)-mediated proliferation of WB F-344 cell. Methods WB F-344 cells were treated with HGF in different concentrations i.e. 0, 20, 40, 80 and 100ng/ml. After incubation for 24 hours, proliferation effect was analyzed with [3H] Thymidine. WB F-344 cells transfected with I?B? plasmid mutants by liposome transfection were used to block NF-?B activity, and then they were treated with 40ng/ml HGF for 0, 15, 30, 60 and 120min, respectively. The nucleoprotein was extracted to determine the NF-?B DNA-binding activity by EMSA. WB F-344 cells were also transiently transfected with BD Great EscA PeTM SEAP vector with liposome and then stimulated with HGF in different concentrations (0, 10, 20, 40ng/ml) for 8 hours to detect the NF-?B transcription activity. WB F-344 cells were pre-treated with NF-?B inhibitor BAY-11-7082 or ASA, and then with 40 ng/ml HGF for 24 hours to analyze the proliferation effect by [3H] Thymidine. Results HGF promoted the proliferation of WB F-344 cells and exhibited positive dose-effect tendency. HGF could enhance the NF-?B DNA-binding activity. NF-?B complexes appeared 15min after addition of HGF. The complexes reached the peak value 30-60min after addition of HGF and then decreased gradually. HGF could also enhance the NF-?B transcription activity, exhibiting positive dose-effect tendency. HGF-mediated proliferation was remarkably inhibited when NF-?B activity was blocked by BAY-11-7082 or ASA. In WB F-344 cells transfected with p-CMV-I?B?, the proliferative response to HGF was completely interrupted. Conclusions HGF may promote the proliferation of WB F-344 cells in a positive dose-effect tendency. The activation of NF-?B is necessary for HGF-mediated proliferation of WB F-344 cells.

In view of the fact that medical inspection equipment sold in the domestic market is mainly imported from abroad and very expensive, we developed a full-automatic fluorescence analyzer in our center, presented in this paper. The present paper introduces the hardware architecture design of FPGA/DSP motion controlling card+PC+ STM32 embedded micro processing unit, software system based on C# multi thread, design and implementation of double-unit communication in detail. By simplifying the hardware structure, selecting hardware legitimately and adopting control system software to object-oriented technology, we have improved the precision and velocity of the control system significantly. Finally, the performance test showed that the control system could meet the needs of automated fluorescence analyzer on the functionality, performance and cost.
Automation, Laboratory ; Equipment Design ; Fluorescence ; Software

Objective:To research the influence and mechanism of Toufeng capsule (TFC) on the concentratioin of blood active substance and single amine neurotransmitter in the brain and blood of magraine mice.Methods: We chose animal disease model of migraine made by hypodermic nitroglycerin and measured the concentration of 5 HT. NE and DA in the rat's brain and blood by means of enzymeimmunoassay experimental methods.Results: TFC could obviously increase the concentration of 5 HT and NE in the bloold and of 5 HT, DA and NE in the brain.Conclusions: TFC might improve neurosystematic regulation functions and enhance the neurotransmitters' secretion and metabolism.

Objective·To investigate the effects of a novel chitosan-silver nitrate gel (CSNG) dressing on anti-septic in wound healing.Methods·By using the ion membrane,the release rate of the new composite materials of silver ion was tested in vitro.Meanwhile,the anti-septic effects of CSNG on methicillin resistant Staphylococcus aureus (MRSA),Escherichia coli and Candida albicans were tested by colony counting.The rat wound healing model was used to detect the ability of new material to kill MRSA in vivo.Results·Compared with control group,the release of silver ions of CSNG was much slower.Sterilization experiment showed that CSNG killed the MRSA,Escherichia coli and Candida albicans much more efficiently,compared to that in the other treatments.Animal experiments also showed that CSNG promoted the rats of wound healing.Conclusion·Combining chitosan with silver nitrate and supplementary material could develop a novel chitosan-silver nitrate gel material,which not only has the obvious effects on antibacterial,but also on promoting wound healing.

Objective Lipid metabolism disorder can lead to male sterility, but its mechanism of affecting spermatogenesis and testis microenvironment remains unclear.This study aimed to investigate the effects of high fat diet on reproductive function and the expression of Ox-LDL within testis in male mice. Methods 16 C57BL/6 mice aged 8 weeks were divided into two groups by random number table method.High fat diet group was fed with high fat diet while normal fat diet group was fed with normal fat diet .At the end of 16 weeks, the levels of TG, TC, Ox-LDL and testosterone in serum were measured.The sperm concentration and motility from caudal epi-didymis were analyzed.The testis structure was observed by HE stai-ning.The localization and expression of Ox-LDL in testis were detec-ted by immunohistochemical technique. Results Compared with the normal diet group mice, the body weight (P<0.05) and the ser-um levels of TC [(2.31 ±0.33)mmol/L vs (6.54 ±0.31)mmol/L, P<0.01], Ox-LDL [(0.32 ±0.03)mg/L vs ( 0.44 ±0.06)mg/L, P<0.01] increased in high fat diet group mice significantly.The serum level of TG was of no significant difference(P>0.05).The serum level of testosterone[(3.64 ±0.43)mg/L vs (0.40 ±0.14) mg/L, P<0.01], the sperm concentration[(9.95 ±0.75)106/mL vs (5.66 ±1.51)106/mL, P<0.01] and the sperm motility[(54.69 ±17.84)%vs (32.48 ±5.80)%, P<0.01] decreased in high fat diet group significantly.HE staining also showed that the amount of Leydig cells, spermatids and spermatozoons reduced obvi-ously in high fat diet group.Immunohistochemistry staining showed that Ox-LDL was mainly distributed around Leydig cells of mice tes-tis.The expression of Ox-LDL in high fat diet group increased significantly ( P<0.01) . Conclusion The expression of Ox-LDL in Leydig cells of high fat diet C57BL/6J mice increases significantly, which may inhibit testosterone biosynthesis and affect spermatogen-ic function.

Objective To evaluate the effect of clinical nursing pathway in emergency treatment of for-eign body in respiratory tract of children. Methods Retrospective analysis was carried out in 555 children with foreign body in respiratory tract from February 2005 to December 2007, who were divided into the control group (271 cases) and the experimental group (284 cases). The experimental group was given managed with standardized clinical pathway,the control group received routine management. The preoperative preparation time, average hospital stay, cost of hospitalization, the extent of mastering of health knowledge by patients'par-ents were observed and the data underwent χ2 and t test. Results All patients of the two groups were cured and no nursing complications happened.The mean preoperative preparation time of the experimental group was 10 hours, which was shorter than that of the control group(27 hours). The average length of hospital stay of the experimental group was (4.13 + 0.81)days, which was shorter than that of the control group ,(6.03 + 1.30) days.The average hospitalization cost of the experimental group was (3498.55 + 269.13) yuan, while the control group was(4246.40 + 977.10) yuan. The extent of mastering of health knowledge by patients' parents, including basic knowledge of the disease, preoperative cooperation knowledge,prevention knowledge and diet knowledge, was superior to that of the control group. Conchusions The establishment of clinical pathway of children with foreign body is conducive to the standardized treatment of the disease, it can shorten the preoperative prepara-tion time, improve the capability of emergency treatment, shorten the length of hospital stay, reducing hospital-ization costs, increase the extent of mastering of health knowledge by patients and their families.

Baculovirus gene expression is the most popular method to make target protein in cultured insect cells. To fast determine the generation of recombinant virus in cultured cells, donor plasmid of pFastBacI was modified by introducing egfp cassette. In the modified vector, egfp cassette was under the control of ie1 promoter, and target gene cassette was under the control of polyhedron promoter. To evaluate the convenience of the genetically modified donor plasmid used in eukaryotic expression, ns1 gene from Bombyx mori bidensovirus was ligated into the donor plasmid to generate recombinant plasmid pFastBacI-[P(ie1)-egfp-sv40]-[P(polh)-ns1-sv40]. Then the plasmid was transformed into DH10B competent cells containing Bm-Bacmid vector to produce the final recombinant Bm-Bacmid with the help of transposase. The resulting recombinant Bm-Bacmid was transfected into BmN cells to generate recombinant virus, which was easily and rapidly judged by green fluorescent signal observed in BmN cells. After infection for 96 h, the BmN cells were harvested and the total protein extracted from the infected BmN cells was subjected to Western blotting analysis. The result showed that a specific protein band about 36 kDa was detected, indicating that NS1 protein was successfully expressed in the BmN cells. In conclusion, the expression of NS1 protein with the modified expression system is useful for further research on the function of NS1 protein.
Animals ; Baculoviridae ; genetics ; Bombyx ; virology ; Cell Line ; Cells, Cultured ; Gene Expression ; Genetic Vectors ; Plasmids ; Promoter Regions, Genetic ; Transfection ; Viral Nonstructural Proteins ; biosynthesis ; genetics

OBJECTIVE: To explore the application of mixed reality technique for the surgery of oral and maxillofacial tumors. METHODS: In this study, patients with a diagnosis of an oral and maxillofacial tumor who were referred to Department of Oral and Maxillofacial Surgery, Peking University School and Hospital of Stomatology from December 2018 to January 2020 were selected. The preoperative contrast-enhanced computed tomography data of the patients were imported into StarAtlas Holographic Medical Imaging System (Visual 3D Corp., Beijing, China). Three-dimensional (3D) model of tumor and key structures, such as skeleton and vessels were reconstructed to three-dimensionally present the spatial relationship between them, followed with the key structures delineation and preoperative virtual surgical planning. By using mixed reality technique, the real-time 3D model was displayed stereotactically in the surgical site. While keeping sterile during operation, the surgeon could use simple gestures to adjust the 3D model, and observed the location, range, and size of tumor and the key structures adjacent to the tumor. Mixed reality technique was used to assist the operation: 3D model registration was performed for guidance before tumor excision; intraoperative real-time verification was performed during tumor exposure and after excision of the tumor. The Likert scale was used to evaluate the application of mixed reality technique after the operation. RESULTS: Eight patients underwent mixed reality assisted tumor resection, and all of them successfully completed the operation. The average time of the 3D model registration was 12.0 minutes. In all the cases, the surgeon could intuitively and three-dimensionally observe the 3D model of the tumor and the surrounding anatomical structures, and could adjust the model during the operation. The results of the Likert scale showed that mixed reality technique got high scores in terms of perceptual accuracy, helping to locate the anatomical parts, the role of model guidance during surgery, and the potential for improving surgical safety (4.22, 4.19, 4.16, and 4.28 points respectively). Eight patients healed well without perioperative complications. CONCLUSION By providing real-time stereotactic visualization of anatomy of surgical site and guiding the operation process through 3D model, mixed reality technique could improve the accuracy and safety of the excision of oral and maxillofacial tumors.
Augmented Reality ; China ; Humans ; Imaging, Three-Dimensional ; Neoplasms ; Retrospective Studies ; Surgery, Computer-Assisted

OBJECTIVETo investigate the role of p38MAPK and PI3K/AKT pathways in mitomycin (MMC)-induced apoptosis in the liver stem-like cell line WB-F344.

METHODSWB-F344 cells were exposed to MMC and apoptosis was evaluated by flow cytometry and DNA fragmentation. Phospho-MAPK and phospho-PI3K/AKT were detected by western blotting.

RESULTSMMC induced apoptosis in WB-F344 cells at 6h after addition of MMC; the maximum level of apoptosis was reached at 24h after MMC exposure. The apoptosis effects of MMC were concentration dependent and inhibited when the PI3K pathway was abolished by the specific inhibitor LY294002, but not inhibited when the p38MAPK pathway was abolished by inhibitor SB203508.

CONCLUSIONApoptosis of WB-F344 cells can be induced by MMC.Although MMC can activate both the PI3K/AKT and p38MAPK pathways, the apoptosis effect of MMC occurs via a PI3K pathway and is not dependent on the p38MAPK pathway.

Animals ; Apoptosis ; Blotting, Western ; Cell Line ; Chromones ; Flow Cytometry ; Mitomycin ; Morpholines ; Phosphatidylinositol 3-Kinases ; Proto-Oncogene Proteins c-akt ; Rats ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases

Background and purpose:Urothelial carcinoma of the bladder (UCB) is the most common cancer in urinary system. Yes associated protein (YAP) gene is closely associated with urothelial carcinoma of the bladder. The study was aimed to explore the effect of siRNA targeting the YAP gene on cell proliferation and migration of T24 cells. Methods:Small interfering RNA (siRNA) was transfected together with LipofectamineTM2000 in T24 human bladder cancer cells to block the YAP signal pathway. The effect of siRNA on cell proliferation and invasiveness was assessed by cell counting kit-8 (CCK-8) assay, Transwell migration assay and wound healing assay. Quantitative real time-Polymerase chain reaction (qRT-PCR) and Western blot analysis were used to conifrm the successful suppression of YAP gene and protein by siRNA. Results:Expression of YAP gene and protein was successfully suppressed after transfected with siRNA which verified by qRT-PCR and Western blot(RNA:F=93.91, P<0.000 1; Protein: F=4.62, P<0.05). As CCK-8 test showed, the proliferation of T24 bladder cancer cells was successfully restrained by inhibition of YAP gene compared with blank control and negative control(12 h: F=6.00, P=0.037;24 h: F=41.72, P=0.000 3;36 h:F=462.8, P<0.000 1;48 h:F=236.6, P<0.000 1;72 h:F=140.5, P<0.000 1). Transwell and wound healing test were performed after YAP gene was interfered by siRNA. The result demonstrated that migration of T24 bladder cancer cells was signiifcantly inhibited (Transwell: F=43.55, P<0.05;Wound healing: F=43.55, P<0.05). Conclusion:This study suggested that YAP gene was an important enhancer for the proliferation and migration of bladder cancer cells.