Cell autophagy is a lysosome dependent degradation pathway which is characterized by cytoplasm vacuoles.It is important to maintain cell internal environment.The abnormal autophagy is associated with oncogenesis closely.The tumor stem cells which rarely exist in tumor tissues have stem cell properties.They can induce tumor generation and also play an important role in the development,metastasis and recurrence of tumor.The regulation of autophagy in tumor stem cells is the research hotspot,which will provide a new theoretical foundation for the clinical treatment of tumors.

The molecular mechanism of autophagy is complicated and it always affects the occurrence and development of tumor in many different degrees,so it is the new hot spot in pathological study area.The angiogenesis includes physiological vascularization and pathological vascularization,and the pathological new vessels which provide nutrition for the continuous growth of tumors have a close relationship to the development and metastasis of tumor.The process of angiogenesis can be affected by regulating the autophagy of vascular endothelial cells,which provides a new method for tumor therapy.

BACKGROUND:It is very nscessary for the prevention and treatment of ostaoarthritis(OA)to understand the tensile mechanical properties of lateral collateral ligaments(LCLs)of knee joints with osteoarthritis,while reports on it are still few.OBJECTIVE:To compare the LCL tensile mechanical property indexes of knee joints between normal animals and OA medels established for this purpose,and to provide qualitative and quantitative referential data about effects of OA on the LCL tensile mechanical properties of animal knee joints.DESlGN,TIME AND SETTING:A randomized controlled trial was finished at the Mechanical Expedmental Center of Jilin University from December 20th to 30th in 2008.MATERIALS:A total of 20 male SD rats of 6 months old were divided into a normal control group and a model group,with 10ones in each group.METHOOS:Rats in the model group were copied into OA models.after which test on tensile was performed to each of the ten rats in the two groups at the speed of 5mm/min on Universal Testing Machine,Shimadzu.Data obtained from the test were processed in polynomial though least square method.MAIN OUTCOME MEASURES:Tensile maximum load.maximum displacement,maximum stress,maximum strain and stress-strain curves.RESULTS:The tensile maximum load,maximum stress,maximum displacement and maximum strain in the normal control group and the model group were(12.754±2.795)N,(27.681±5.832)MPa,(2.754±0.707)mm,(11.679±2.373)%and (7.183±1.817)N,(16.162±3.403)MPa,(1.827±0.768)mm,(8.019±2.811)% respectively,from which we could see that each index in the normal control group was greater than that in the model group obviously (P＜0.05).CONCLUSION:The tensile maximum load,maximum stress,maximum displacement and maximum strain all decreased after OA,which indicates that osteoarthritis has certain effects on tensile mechanical properties of LCLs of knee joints as well as stress relaxation.

Objective To establish a quality standard for Aiweixin oral liquid. Methods TLC was applied to the identification of Caryophylli Flos and Crocin in Aiweixin oral liquid. Eugenol was analyzed by GC. Results TLC of Caryophylli Flos and Crocin had distinct separation of characteristic spots and there was no interference in negative comparison. The linear response ranges of Eugenol were between 0.052 17-2.086 8μg (r=0.999 9). The samples were steady within 24 h. The average recovery of eugenol was 95.7%, RSD=1.4% (n=6). Conclusion The method is simple, accurate and reliable, and can be used for the quality control of Aiweixin oral liquid.

Specimens of skin, skeletal muscle, adipose tissue, brain, peripheral nerve and catilage were taken from 6 cadavers (divided into 2 groups) perfused and embalmed with formalin. 90 pieces of the above-mentioned tissues were taken in different times and the paraffin sections were stained by routine HE. Electron microscopic observation was made from some skin and skeletal muscle specimens, and histochemical methods were used to demonstrate DNA, RNA, keratin(-SH, -SS), fatty acids and calcium in some skin materials. Loyez stain method for myelin sheath was applied to some brain and nerve sections.1. Skin tissue: In both group A and B, 83.3% and 78.9% of skin materials were appraised as grade I and II (excellent and good) respectively. The morphological changes appeared mainly as karyopyknosis of the epidermal cells and shrinkage of the secretory part of the sweat glands. Most of the skin tissues of the corpses perfused 3～4 hrs after death were well preserved, only 8.3% being in grade Ⅲ (moderate). On the other hand, 43% of skin materials taken from those perfused 30 hrs after death were in grade Ⅲ. Ultrastructurally, the shape of the epidermal cells and their adhensions still remained intact, but karyopyknosis or chromolysis and autolysis of most of the cytoplasmic membraneous structures were found. In those cadavers preserved for less than one year, there was no evidence of progressive changes in microscopic and submicroscopic structures of the skin. The tonofillaments and desmosomes were still kept well in those specimens preserved for many years, and their presence may be the cause of maintaining cell integrity. Histochemically, mercapto-reaction of keratin was kept stable and found to be well shown even in old samples of skin, but the disulfide reaction usually showed marked fading with time, probably due to exofolition of the stratum corneum. However, this reaction was well shown in the hairs and not weakened for many years.2. Fat tissue: Hydrolysis of lipid substances of subcutancous fat and sebaceous gland, as shown by the presence of fatty acid, was found in the corpses preserved even as short as half a year.3. Skeletal muscle: The preservation state of the skeletal muscle was also dependent on the complete perfusion in time. The structure of the muscle fibers was generally worse preserved than that of the skin as shown in severe myolysis. Since stagnation of the small blood vessels frequently accompanied myolysis, obstruction of microcirculation which prevented the permeation of fomalin into the muscle tissues may be the cause of rapid myolysis. The ultrastructural changes of the muscle fibers were the same as those of the epidermal cells. In all specimens even as old as 30 years, two types of myofillaments were well preserved, and bands could be seen distinctly.4. Nervous tissue: The brain tissues were well preserved in those corpses that were perfused shortly after death, but the peripheral nerves usually showed severe autolysis, obviously it is due to the same reasons as those for myolysis.5. Cartilage: The structural changes of cartilage after death were slow and all specimens of cartilage were well preserved.

Objective:To observe the expression of BTLA on T cells during activation and further analyze its inhibitory effects on T cell activation in different phases.Methods: T cells from PBMC were enriched by negative selection using magnetic beads.Expression of BTLA,CTLA-4 and PD-1 on freshly isolated human T cells and kinetics expression of BTLA,CILA-4 and PD-1 on CD3 mAb stimulated T cells were examined by flow cytometry.T cells were stimulated by anti-CD3 mAb combined with anti-CD28 mAb in the presence of anti-BTLA mAb 8H9,then T cell proliferation was tested by MTT assay in the different culture time.Immature DCs were generated from monocytes cultured in the medium containing GM-CSF and IL-4, and further driven to maturation by anti-CD40 mAb.Expression of HVEM on DCs was measured by flow cytometry.T cells were co-cultured with DCs in the presence of soluble 8H9 or anti-HVEM antibody to block HVEM-BTLA interaction,T cell proliferation was measured by MTT assay.Results:Freshly isolated T cells exhibited high levels of BTLA expression, but not CTLA-4 and PD-1.After T cell activation, BTLA expression decreased on first 2 days, with rapidly increasing to high levels.Unlike BTLA, expression of CTLA-4 and PD-1 was gradually increased during T cell activation.8H9 significantly inhibited the proliferation of T cell stimulated by CD3 mAb and CD28 mAb.8H9 could still exhibit inhibitory effect on T cell proliferation after 24 h or 48 h of preactivation by CD3 mAb plus CD28 mAb stimulation.HVEM was highly expressed on immature DCs, and down-regulated on mature DCs.Blockade of BTLA by soluble 8H9 or anti-HVEM antibody enhanced DC-mediated T cell proliferation within 48 h.Conclusion: BTLA signal enhances the threshold of T cell activation and plays importantly negative regulatory role in the initiation and early phase of T cell activation.

ObjectiveTo explore the effects and underlying mechanisms of acid sensing ion channels (ASICs) on pain behavior in a rat model of post-incision pain.MethodsFifty-eight adult male Sprague Dawley rats were used in this study,four rats were used for immunofluorescence test,thirty rats were employed for pain behavior test,and twenty-four rats were used for Western blot.Rats used for pain behavior test and Western blot were randomly divided into 3 groups:control group ( C group),incision pain model group ( I group) and amiloride group (A group).Plantar skin of rats in A group were infiltrated with 20 μl(200 μg)amiloride solution.Paw withdrawal mechanical threshold(PWMT) and paw withdrawal thermal latency(PWTL) of all rats in pain behavior test was tested at 24 h preoperative,2 h,4 h,8 h,12 h,24 h postoperative.Western blot was tested at 4 h postoperative.ResultsImmunofluorescence test displayed ASIC3 was expressed in plantar skin of all rats.The basal level of PWMT and PWTL of all rats in three groups was C group( (23.15 ± 5.10) g,( 11.32 ± 1.21 ) s),I group ( (23.26 ± 5.69) g,( 11.75 ± 2.01 ) s),A group ( (23.63 ± 4.96 ) g,( 11.47 ± 1.96) s) respectively,which was no significantly difference (P ＞ 0.05 ).PWMT and PWTL of I group and A group was significantly lower than that of C group at all time points postoperative (P ＜ 0.05) ; PWMT and PWTL of A group was at 2 h( ( 13.75 ±3.25)g,(9.96±1.32)s),4h((14.05±3.75)g,(9.17±2.11)s),8 h((9.75 ±2.74)g,(8.11 ±1.22)s)postoperative,which was significantly higher than that of I group (P ＜ 0.05 ).Compared with that of C group,the level of pERK1/2 expression was significantly increased in I group at 4 h postoperative (P ＜ 0.05 ),which could be inhibited by amiloride local infiltration (P ＜ 0.05 ).ConclusionASIC3 can mediate incision pain in a rat model of post-incision pain,through pERK1/2 signaling pathway,which can be inhibited by amiloride.

Objective　In order to find out influencing factors of corrosiveness of common chemical disinfectants used at present for medical instruments on varied metal material, and to offer scientific bases for working -out corresponding state standards. Methods　Liquid instruments disinfectants containing chlorine and glutaraldehyde compound disinfectant were chose to take an example to study their corrosiveness to varied sorts and type s of the metal. Sing le related factors were researched by contrast test study in the process of dete rmining metal corrosion rate (R value, mm*a-1) citing from GB10124-88 me thod. Results　Varied chemical disinfectants had different metal R valu e. R value was in fluenced by kinds of metal pieces, sorts of soaked vessel material, drying tempe rature, whether changing liquid disinfectants or not and water quality, while d ifferent capacity of balance measure had no effect on R value. Concl usi ons　R value was affected by kinds and types of metal pieces and other factors. These fac tors should be considered sufficiently while determining or comparing corrosiven ess of chemical disinfectants to soaked metal.

Objective To investigate the different distribution of regulatory T cells (Tregs) and CD8+T cells in the local immune microenvironment of mucosal malignant melanoma and cutaneous malignant melanoma, and analyze the relationship between the two indicators and the prognosis of patients. Methods Immunohistochemistry staining was used to assess the ex?pression of Foxp3+Tregs and CD8+T cells in tumor microenvironment of 58 patients with malignant melanoma. The correlation between two factors, clinicopathological characteristics, and prognosis were analyzed. Results There is no correlation be?tween the expression of Foxp3 and CD8. The number of Foxp3+Tregs was significantly higher in mucosa malignant melanoma than that in cutaneous malignant melanoma (t=2.648, P=0.011). The proportion of Foxp3highTregs was significantly higher in pa?tients with tumor diameter≥3 cm, lymph node metastasis and distant metastasis than that in patients with tumor diameter<3 cm, no lymph node metastasis and no distant metastasis (P<0.05). In addition, in patients with ulceration that proportion was significantly higher in CD8high group than that in patients without ulceration (33.3%vs 5.9%, P<0.05). The median progres?sion-free surial (PFS) was 12 months in Foxp3high group, which was significantly longer than that of Foxp3low group (31 months, P<0.05). The median PFS was significantly higher in CD8high group (25 months) than that of CD8low group (12 months, P<0.05). Subgroup analysis showed that the median PFS was 7 months in Foxp3high CD8low group, which was significantly lower than that of Foxp3highCD8high group (25 months) and Foxp3lowCD8low group (18 months, P=0.003). Univariate analysis showed that median PFS was different in patients with different tumor location, different number of Foxp3+Treg, different number of CD8+T cells, and distant metastases. Conclusion The number of Tregs is closely associated with metastasis in patients with malig?nant melanoma. Compared with cutaneous malignant melanoma, our results indicate that the poor prognosis of mucosal malig?nant melanoma may be associated with the infiltration of more Tregs.

Objective To observe the different radiosensitivity induced by different doses of 137Csγ-ray irradiation between hematopoietic stem and progenitor cells. Methods Seventy-two C57BL/6 mice were randomly divided into control group and irradiated groups (2, 4 and 6 Csγ-ray irradiation, n=18 for each group). Mice of control group received sham irradiation, and the rest accepted 2, 4 and 6 Gy137Csγtotal body irradiation, respectively. After 14-day, 35-day and 56-day irradiation, the peripheral blood samples were collected by balls enucleation. The number of bone marrow nuclear cells, hematopoietic stem and progenitor cells were counted. Results The peripheral blood of irradiated mice showed significant changes in the number of white blood cells (WBC), red blood cells (RBC), platelets (PLT) and hemoglobin (HGB) in a dose-response relationship. Compared with the control group, the numbers of BMNCs and hematopoietic progenitor cells (HPCs) were significantly lower in irradiated group. At 35 d and 56 d after 6 Gy irradiation the numbers of BMNCs and HPCs were significantly lower than those of control group (P<0.05). There were no significant differences in numbers of BMNCs and HPCs between irradiated groups (2 and 4 Gy) and control group. The number of bone marrow hematopoietic stem cells (HSCs) was significantly lower in irradiated group than that in control group after 14-d and 56-d irradiation (P<0.05). Conclusion 137Csγ-ray irradiation has some damage in mouse hematopoietic system. The damage caused by radiation is persistent to hematopoietic stem cells.