It is the current confusion encountered by integrated Chinese and Western medicine that how to find the breakthrough direction of integrating Chinese and Western medicine， from crossover to integration to innovation， and open up a new horizon of integrated Chinese and Western medicine. The progress of Chinese medicine lay in expanding the scope of diagnosis and treatment with the help of modern diagnostic and therapeutic equipments and developing “micro” identification， while the progress of Western medicine lay in looking at “macro” and developing systemic medicine and integrated medicine， both of which are in the direction of each other. The “state-target identification and treatment” may become an important way to build a modern diagnosis and treatment system of integrated Chinese and Western medicine， and the thinking mode of “from target to state” is a further refinement and development on the basis of the theoretical system of “state-target identification and treatment”， which provided a clearer solution for the current stage of the integrated Chinese and Western medicine model， and pointed out the important development direction for the future integrated Chinese and Western medicine. From the perspective of strategic level and diagnosis and treatment practice， it integrated the “target-state” thinking mode into the modern diagnosis and treatment model of the integrated Chinese and Western medicine， i.e.， “Western medicine as the basis and treating with Chinese medicine； Chinese medicine as the basis and treating with Western medicine”. On the one hand， Western medicine should strengthen the reference to the traditional theories and holism of Chinese medicine， and advocate a higher level of education on the integrated Chinese and Western medicine under the guidance of the traditional theories of Chinese medicine. On the other hand， the “from target to state” mode of thinking should be applied to guide the establishment of diagnostic and treatment strategies and clinical selection of medicines in clinical practice， so as to locate the target and adjust the body state in a gradual and orderly manner， and to provide practical methods for the modern clinical work of the integrated Chinese and Western medicines. Chinese and Western medicine systems can learn from each other， combine organically， give full play to their respective strengths， and form an internal law， so as to make breakthroughs and innovations in the integrated Chinese and Western medicine model.

OBJECTIVES: Liver cancer is the sixth most common malignant tumor in the world. Hepatocellular carcinoma (HCC) accounts for 85%-90% of all patients with liver cancer. It possesses the characteristics of insidious onset, rapid progression, early recurrence, easy drug resistance, and poor prognosis. NIMA related kinase 2 (NEK2) is a cell cycle regulating kinases, which regulates cell cycle in mitosis. Cellular senescence is a complex heterogeneous process, and is a stable form of cell cycle arrest that limits the proliferative potential of cells. This study aims to investigate the relationship between the expression level of NEK2 and the senescence in hepatoma cells, and to explore the effect of NEK2 expression on hepatoma cell senescence and the underlying molecular mechanism. METHODS: A total of 581 senescence-relevant genes were obtained from the GenAge website. The gene expression data of tumor tissues of 370 HCC patients were downloaded from the Cancer Genome Atlas database. The co-expression of NEK2 and aging-related genes was analyzed by R-package. KEGG was used to analyze the significant gene enrichment pathway of differentially expressed genes in NEK2 overexpression HEK293. The stable transfected cell lines with overexpression and knockdown of NEK2 were constructed in hepatoma cell line SMMC-7721 and HepG2, and senescence-associated β-galactosidase (SA-β-gal) staining was used to detect senescence, the cell proliferation was detected by CCK-8 method and clone formation experiment, the cell cycle was analyzed by flow cytometry, and the expression of proteins related to p53/p21, p16/Rb, and phosphatase and tensin homolog deleted on chromosome ten (PTEN)/Akt signal transduction pathway was detected by Western blotting. RESULTS: There were 320 senescence related genes co-expressed with NEK2. KEGG analysis showed that the senescence signaling pathway was significantly enriched in HEK293 cells with overexpression of NEK2.Compared with SMMC-7721 or HepG2 without knockdown of NEK2, the senescent cells of SMMC-7721 and HepG2 with knockdown of NEK2 were increased, cell proliferation and clone formation were decreased significantly, the percentage of cells in G₀/G₁ phase was increased, the expression levels of phospho-Akt (p-Akt) and phospho-Rb (p-Rb) protein were decreased significantly, and the expression level of p16 protein was increased significantly (all P<0.05). Compared with SMMC-7721 or HepG2 transfected with blank plasmid, the senescent cells of SMMC-7721 and HepG2 overexpressing NEK2 were decreased, the cell proliferation and clone formation were increased significantly, the percentage of cells in G₀/G₁ phase were decreased, the expression levels of p-Akt and p-Rb protein were increased significantly, and the expression level of p16 protein was decreased significantly (all P<0.05). CONCLUSIONS NEK2 may mediate the anti-aging effect of hepatoma cells through p16/Rb and PTEN/Akt signal transduction pathways, which provides a new theoretical basis for NEK2 to promote the progress of liver cancer and a new idea for the targeting treatment for liver cancer.
Carcinoma, Hepatocellular/pathology* ; Cell Line, Tumor ; Cell Proliferation/physiology* ; Cellular Senescence/genetics* ; HEK293 Cells ; Humans ; Liver Neoplasms/pathology* ; NIMA-Related Kinases/genetics* ; Proto-Oncogene Proteins c-akt/metabolism*

Objective To explore the feasibility of needle-embedding therapy in the treatment of chronic myocardial ischemia using a miniature pig model established by placement of an Ameroid constrictor at the left anterior descending branch (LAD) of coronary artery in Bama miniature pigs during surgery. Methods The miniature pig model of chronic myocardial ischemia was established by placement of an Ameroid constrictor at the left anterior descending branch of the left coronary artery in Bama miniature pigs. The pig models were randomly divided into the treatment group (the"Neiguan " group) and the control group (the "Zusanli " group), and were treated with needle- embedding electroacupuncture at the"Neiguan" (PC6) and "Zusanli" (ST36) acupoints, respectively. Myocardial samples were taken at 6 weeks after surgery for light and electron microscopic examinations. Results Gross pathology showed that ischemic area in the myocardium appeared in both experimental groups. The ischemic area in the "Zusanli "group was larger than that of the"Neiguan"group. Histopathology showed that the acupuncture treatment at the"Neiguan"acupoint reduced the ischemic injury in the pig myocardial tissues. Ultrastructural observation of the myocardium showed mitochondrial vacuolization in cardiomyocytes and myocardial fibrosis in both groups. Conclusions Acupuncture therapy at the"Neiguan"acupoint of pericardial channel may exert protective effect on the myocardial ischemia by reducing the ischemia-injury of cardiomyocytes, but can not inhibit the already existed ischemia-induced cardiomyocytic injuries. Our findings suggest that the establishment of miniature pig model of chronic myocardial ischemia by surgically placing an Ameroid constrictor on the left anterior descending branch of left coronary artery and the needle-embedding in acupoints is feasible for the treatment of chronic myocardial ischemia in this pig model.

Objective To investigate the effect of TiO2 nanotube arrays covalently modified by recombinant human bone morphogenetic protein-2(rhBMP-2) on the early bioactivity of mesenchymal stem cells(MSC) in vitro and to provide experimental evidence for the biochemical modification of titanium implants.Methods In the experiment group,double titanium nanotube arrays were prepared by anodization,and were chemically grafted with rhBMP-2.Mechanically polished pure titanium was used as blank control group,and titanium dioxide nanotubes was used as negative control A group,and titanium dioxide nanotubes + carbonyldiimidazole as negative control B group.Field emission scanning electron microscope(FE-SEM) and X-ray photoelectron spectroscopy(XPS) were used to detect the morphology and physicochemical properties of the experiment group,blank control group and the negative control group.Cell adhesion on the specimen surface of the experiment group,blank control group and negative control group on the 1st day was tested.Cell proliferation on the 1st,3rd and 5th day and alkaline phosphatase activity on the 5th,7th and 11th day was also tested.Results FE-SEM showed that the surface of titanium nanotubes loaded with rhBMP-2 possessed visible miliary particulate matter.XPS showed that nitrogen peak in the group of titanium nanotubes loaded with rhBMP-2 was significantly greater that those in the other groups.FE-SEM showed that the cells on the surface of the experimental group on the 1st day spread well,better than those in the control group and negative control group.Cell proliferation activity on the 1st day in different groups was not obvious(P＞0.05),the A value of the experimental group on the 3rd and 5th day (3.295±0.153,3.823±0.059) were significantly higher than those in the control group(2.479±0.064,3.131±0.096) and negative control A group(2.715±0.075,3.371±0.047) and negative control B group(2.756±0.132,3.637±0.047)(P＜0.05).Alkaline phosphatase activity on the 5th,7th and 1 1th day in the experimental group (0.0477 ± 0.0287,0.0615 ± 0.0016,0.0667 ± 0.0018) were better than those in the control group,negative control A group and negative control B group(P＜0.05).Conclusions Titanium nanotube arrays can be loaded with rhBMP-2 by biochemical methods and have good biocompatibility.

Objective:To evaluate the effect of different polishing instruments on the microstructure of composite resin.Methods:Specimens of 5 kinds of resins were randomly divided into three groups and were polished with Astropol(group A),Sof-Lex(group B) and Super-snap(group C)respectively(n=3 for each resin).Specimens of natural enamel surface were used as the controls(group D).The surface roughness(Ra)and microstructure of the specimens were evaluated by an atomic force microscope(AFM).Results:Ra value in group A was lower than that in group B(P=0.015);Z350 XT resin obtained the lowest Ra value among the 5 kinds of resin specimens(P<0.05).All of the resin specimens had similar average Ra after polishing procedure(P>0.05).Conclusion:Astropol polishing instrument is better than Sof-Lex;Z350 XT has better polish performance.The three polishing instruments are ef-fective in polishing the 5 composite resins.

Doppel protein (abbreviation Dpl) is a newly recognized Glycosyl phosphatidyl inositol (GPI) anchored and highly glycosylated protein, which is similar to prion protein (PrP) in the chemical structure. The encoding gene of Dpl named PRND locates at the downstream of the prion protein gene (PRNP). These two proteins are different in physiological functions. The expression of Dpl focuses on testis tissue at the adult, and takes an important role in maintaining sperm integrality, normal fertility, and motion ability. We reviewed the biological characters, physiological functions of Dpl and its effects on male reproduction in order to provide theory guidance for the study on physiological function and male reproduction controlling.
Animals ; GPI-Linked Proteins ; Humans ; Male ; Prions ; metabolism ; physiology ; Reproduction ; physiology ; Testis ; growth & development ; metabolism

Objective To investigate the expression of Nogo-A in the retinal ganglion cells (RGCs) and their axons of mouse embryos and its time course changes. Methods Sections of retinofugal pathway of C57 mouse embryos at different developmental stages were immunostained with Nogo-A specific antibody and observed by a confocal microscopy. The identity of Nogo-A positive cells was partially revealed by double-staining together with Tuj-1. Results At the early stage of E12, Nogo-A was densely expressed in some radially-orientated cells in retina. The immunopositive signals appeared in the cytoplasm, on the cell membrane and axons. The double-labeling together with Tuj-1, a neuronal marker, showed that nearly all the RGCs and their axons expressed Nogo-A protein. At the later stage of E13, the number of Nogo-A positive neurons in retina decreased dramatically. And those Nogo-A positive RGCs were specifically located in the ventricular part and the ciliary margin zone of the retina. At this stage, only a very few axons maintained their Nogo-A expression in the fiber layer of the retina, while most lost their Nogo-A distribution. When most RGCs had fully differentiated at E15, there was no detectable Nogo-A immunopositive staining in the retina and only a few retinal fibers were Nogo-A immunopositive. The similar expression patterns of Nogo-A was found in a few axons along the optic disc, optic stalk, optic chiasm and optic tract. Worthy of note, the retinal axons with Nogo-A distribution in the optic tract were exclusively found in the superficial area, where the newly-arrived axons were traveling through during development. Conclusion The expression pattern and its time course change suggested that Nogo-A was an important protein expressed by the newly differentiated RGC neurons and their projecting axons, whilst the mature RGCs down-regulated their expression. Nogo-A in the new born RGCs might play some cell-intrinsic roles such as decreasing axon branching in vivo.

Objective To study the expression of Livin and its relalionship with expression of p53 in small cell hmg cancer(SCLC).Methods Immunohistochemical S-P method was used to detect the ex-pression of Livin and p53 protein in 30 SCLC tissues and 16 para-cancerous lung tissues.Results Livin protein was expressed in 17 of 30 SCLC tissues(56.67%),but Livin protein showed low levels in para-can-cerous lung tissues(12.50%),P＜0.01.There was no significant correlalion between positive Livin protein expression and age,sex,TNM staging,lymph node metastasis and lumor diameter(P＞0.05).p53 protein was expressed in 14 of 30 SCLC tissues(46.67%),but p53 protein was no expressed in para-caneerous lung tissues,P＜0.01.The expression of Livin protein was positively related to the expression of p53 protein(P＜0.01).Conclusions The aberrant expression of Livin may be a new target for diagnosis and gene treatment of SCLC.The aberrant expression of Livin and p53 may play synergetic role in process of carcinogenesis of SCLC.

BACKGROUND: The environmental change of regeneration of peripheral nerve fiber can accelerate the regeneration of nerve fiber and the recovery of nerve function. Physiotherapy can improve the local microcirculation of the injured area, and promote and stimulate the recovery course of trauma. Therefore, whether choosing different physiotherapies and controlling irradiation dose can speed up the regeneration of nerve fiber?OBJECTIVE: To observe the effects of different physiotherapies in promoting the regeneration of injured nerve fiber.DESIGN:Randomized grouping and controlled animal experiment.SETTING: Laboratory of Anthropotomy and Histo-Embryology Department,Peking University Health Science Center.MATERIALS: The experiment was conducted in the Laboratory of Anthropotomy and Histo-Embryology Department, Peking University Health Science Center, from September 2001 to September 2002. Fifteen adult male SD rats, weighting 185-220 g, were selected.INTERVENTIONS: The model of sciatic nerve injury was established and randomly assigned to 3 groups. Bio-spectrum group and infrared group with 5rats in each received the corresponding physical irradiation. Injury control group was given no irradiation. The slices of sciatic nerve at the operation side of the animals in the three groups were collected on day 14 after operation. The degeneration rate of the injured nerve fiber was observed under the optical microscope to make indirect assessment of the protective effect of physiotherapy after the nerve was injured.MAIN OUTCOME MEASURES: The degeneration rate of nerve fibers at the breakpoint and at 4 points near the spinal cord ( 1,2,3,4 mm) and away from the spinal cord( 1,2,3,4 mm).difference in the degeneration rate of nerve fibers at the breakpoint away from the spinal cord in bio-spectrum group and infrared group as compared with the degeneration rate of nerve fibers in bio-spectrum group(68% ) and infrared group(89% ) was obviously reduced and that in control group was the the spinal cord in the 3 groups showed decreasing trend: the closer to the spinal cord, the fewer the degenerative nerve fibers. The number of degenerative nerve fibers was the smallest in bio-spectrum group, second smallest in infrared group, and the largest in control group.CONCLUSION: The assisting physiotherapy can decrease the number of regenerative nerve fibers, thus promoting and accelerating their recovery. Irradiation with bio-spectrum equipment is more effective.

Objective To study the distribution of Nogo-A in the retina and the changes after the optic nerve(ON) injury in hamsters. Methods In this experiment,ON was crushed at 2?mm behind of the eyeball.After 3?d,5?d and 7?d post-axotomy,the Nogo antiserum immunoreactive staining on section of the retina was performed. Results Nogo was expressed at every layer of the retina with the strongest expression on 3?d post-axotomy.The numbers of Nogo-A positive RGCs decrease as the survived time increased.Conclusion Nogo-A in the retina is not unique secretion from the neuroglia.The change in the distribution and level of expressed of Nogo-A in the retina is correlated with time advancement after injured of ON.