Autophagy contributes to the pathology of various neurodegenerative conditions.Rapamycin,an agent against Candida albicans or immunosuppressive,was found to be effective against tumor.Furthermore,it also induces autophagy by inactivating the protein mammalian target of rapamycin(mTOR),FKBP and VMP1,thus could offer a tractable therapeutic agent for neurodegenerative diseases.

Objective To evaluate antibacterial and antifungal effect of the composite chitosan artificial skin in vitro. Methods The standard strains were used in the experiment, including Staphylococcus aureus (ATCC25923), Escherichia coli (ATCC25922), Pseudomonas aeruginosa (ATCC27853) and Candida albicans (ATCC10231). Twelve agar plates were prepared for each standard strain, which were equally divided into the trial and the control groups. In the trial groups, 50 ?L composite chitosan dermal substitute was added to each prepared agar plate, two samples for each plate. In the control groups, composite collagen-gelatin dermal substitute was used. After the plates were incubated at 35 ℃ for 18 ~ 24 h, the antibacterial or antifungal rings of every sample were measured. Results The composite chitosan dermal substitute showed the antibacterial effect on Staphylococcus aureus (ATCC25923), Escherichia coli (ATCC25922), and Pseudomonas aeruginosa (ATCC27853) (P

Objective To rapidly isolate and culture sweat gland epithelial cells in vitro and to observe the characteristics of the cells. Methods The secretory coils of sweat glands were dissected and picked out under an anatomical microscope, then digested by collagenase. The harvested epithelial cells of sweat gland were observed for their growth characteristics and identified by immunohistochemistry. Results The cultured epithelial cells grew very well. About 3 weeks later, a distinctive cobblestone appearance was observed in the culture. The antibody-staining showed the cells were positive for epithelial membrane antigen and cytokeratin, but negative for actin, which confirmed that the cells were sweat gland epithelial cells. Conclusion A method is established for rapid isolation and culure of sweat gland epithelial cells in vitro.

AIM: To observe the inhibitory effect of recombinant human endostatin (rhES) on plaque angio-genesis, and to explore the regulatory mechanism of Dll4 /Notch pathway in the anti-angiogenic effect of rhES.METH-ODS: Male Wistar rats were randomized into 3 groups: normal control group (N group), atherosclerotic model group (AS group), and rhES treated group (AS +rhES group).The rats in N group were fed a normal diet, while the remaining 2 groups were established to atherosclerotic rat model via high-cholesterol diet, intraperitoneal injection of vitamin D3 and aor-tic balloon injury.The rats in AS +rhES group received intraperitoneal injection of rhES.The blood total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), C-reactive protein (CRP), interleukin-1 (IL-1) and troponin I (TnI) were measured.The atherosclerotic abdominal aortas were taken for pathological observation.Immu-nohistochemical staining was used to measure the density of neovessels in the plaques, which were marked by CD31.The protein levels of Dll4 and Notch1 in the aortas were analyzed by Western blot.RESULTS: The levels of blood TC, TG, LDL-C, CRP and IL-1 in AS group and AS +rhES group were much higher than those in N group (P <0.05), and no sta-tistical difference between AS group and AS +rhES group was observed.The expression of CD31 in AS group was the high-est among all groups.Compared with AS group, the density of neovessels in the plaques of AS +rhES group decreased sig-nificantly (P <0.05).The protein expression of Dll4 and Notch1 in AS group was lower than that in N group (P <0.05). Compared with AS group, the protein expression of Dll4 and Notch1 increased significantly (P <0.05).CONCLUSION:rhES has the ability to inhibit plaque angiogenesis in rats.The activation of Dll4 /Notch pathway may be the mechanism of rhES in inhibiting plaque angiogenesis.

AIM: To evaluate effects of diltiazem on platelet hyperreactivity in situations associated with endothelial injury and their possible relationship to cytosolic calcium concentration. METHODS: Blood samples were collected at 7 time points from 35 patients undergoing percutaneous transluminal coronary angioplasty (PTCA) who received combined diltiazem and aspirin/ticlopidine therapy or aspirin/ticlopidine therapy alone. Platelet expression of glycoprotein Ⅱb/Ⅲa and cytosolic calcium concentration were measured, respectively, by whole blood flow cytometry and fluorospectrophotometry. The effects of diltiazem of different concentrations on expression of glycoprotein Ⅱb/Ⅲa were also studied in vitro in blood samples from patients with chronic stable angina. RESULTS: Of the two treatments, aspirin/ticlopidine therapy did not prevent an acute increase of expression of glycoprotein Ⅱb/Ⅲa 5 minutes and 10 minutes after first inflation and 10 minutes after PTCA, whereas combined diltiazem and aspirin/ticlopidine therapy had a significant inhibitory effect. In the group receiving aspirin/ticlopidine therapy, there was a short-term elevation of platelet [Ca~(2+)]i immediately following PTCA which was significantly reduced by diltiazem treatment. Expression of glycoprotein Ⅱb/Ⅲa was significantly inhibited in vitro by diltiazem in the concentration of 200 ?g/L or higher, but not 50 ?g/L. CONCLUSIONS: Combined diltiazem and aspirin/ticlopidine therapy significantly inhibited platelet activation that continued in the presence of conventional aspirin/ticlopidine treatment. Antiplatelet effects of diltiazem were probably a consequence of reduction of platelet [Ca~(2+)]i and may only be achieved in higher than therapeutic concentrations. [

Purpose To explore the Relationship between expression of apoptosis-modulating proteins amdmultidrug resistance in K562/VCR cells. Methods Irnmunocytochemical methol and western blot wereused to analyze the expression of apoptosis-modulating proteins (Bcl - 2, Bcl-XL, Bax, Bak ) in multidrugresistant cell line K562/VCR and drugsensitive cell line K562. Results The positive cell rates ofapoptosis-suppressing protein Bcl-2 and Bcl-XL in K562/VCR were (40.0 ± 8.0) % and (60.0 ± 10.0) % .While the rates in K562 were (1.0 ± 0.3) % and (20.0 ± 4.0) %. There was significant difference in thepositive cell rates of Bcl - 2 and Bcl - XL between K562/VCR and K562 ( n = 3, P ＜ 0.05 ). It was alsofound there was no significant difference in expression of Bax between K562/VCR and K562. Furthemore,Bak was not expressed in both K562/VCR and K562 or the expression was very low. Conclusions Wesuggest that Bcl-2 and Bcl-XL play important roles in multidrug resistance in K562/VCR, while Bax and Bakmight not be important.

Objective To investigate the effect of hepatocyte growth factor (HGF) on migration and apoptosis of,as well as phosphorylated-Akt (p-Akt) expression in cultured human eccrine sweat gland epithehal cells (hESGc).Methods The first generation of hESGc were cultured in keratinocyte serum free medium (KSFM) and treated with various concentrations (2,20,40μg/L) of HGF for different durations.Then,cell scratch test was performed to detect cell migration,a double staining flow cytometry assay using annexin VFITC/propidium iodide to detect cell apoptosis.and Western blot to measure the expression of p-Akt.Results HGF of 2μg/L had no effect on the migration of hESGc,while that of 20 μg/L and 40μg/L could promote the migration of hESGc by 33.2% and 228.2%.respectively.The average number of cells migrating into the scrach zone was significantly lower in untreated cell group than that in 20 and 40μg/L HGF-treated cell group (17.3±5.5 vs 23.0±6.3 and 56.7±7.9,t=2.653, 15.858,P<0.05,0.01, respectively).The apoptosis rate was 14.76% in untreated cells,14.16%,13.5% and 8.87% in cells treated with HGF of 2,20 and 40μ/L, respectively;there was a significant difference between untreated cells and 40μg/L HGF-treated cells (t=7.852,P<0.01).HGF could activate the phosphorylation of Akt protein and increase the expression of p-Akt.Conclusion HGF could promote the migration of,inhibit the apoptosis of,and stimulate the p-Akt expression in.hESGc.

Objective To investigate the clinical outcomes of surgical excision combined with recom binant interferon aipha-2b in the treatment of acral malignant melanoma (MM). Methods Fifteen patients with acral MM admitted to the department since 2004 were recruited into this study. The tumors varied from 1.8 mm to 3.9 mm in invasion depth. Thin tumors with an invasion depth of 1.8 - 2.0 mm were excised with a margin of lcm beyond the tumors, and those with an invasion depth of 2.0 - 3.9 mm were excised with a margin of 2 cm beyond the tumors. After excision, 4 cases of minor excision were sutured directly, 10 cases of large excision were repaired with adjacent skin grafts and flaps, 1 patient with the involvement of the first metatarsophalangeal joint underwent toe amputation followed by the repair of planta wound surface with the remaining skin on the dorsa of toes. Patients received intramuscular recombinant interferon alpha-2b for 3 months (3 million units daily for the first 3 days and 6 million units for the remaining days) following operation. Results There were 6 cases of MM in situ and 9 cases of invasive MM in this study. All the skin grafts and flaps survived. Within the 3-year follow up, relapse was observed only in 1 patient with invasive MM. Recovery was achieved in the functions of feet in all patients. Conclusion The excision of tumors with a margin determined by tumor thickness plus intramuscular interferon alpha-2h may improve the survival of patients with cutaneous MM in planta pedis with avoidance of amputation.

Adult ; Aorta, Thoracic ; injuries ; Coronary Vessels ; injuries ; Humans ; Male ; Tomography, X-Ray Computed

OBJECTIVETo investigate the practicability of repair of full skin loss of rabbits with composite chitosan artificial skin.

METHODSDermal substitute was prepared aseptically by mixing fibroblasts with composite dermal matrix gel. Keratinocytes were then seeded on the substitute and submersion - cultured thereafter for 1 week in keratinocyte culture medium. The composite was further cultured for 1 approximately 2 weeks on the surface of the culture liquid to form artificial skin. The composite chitosan artificial skin was then grafted onto the full skin loss wound of rabbits. Histological changes were undertaken periodically by tissue sampling from the grafted wound. The systemic reaction of rabbits to the artificial skin was observed.

RESULTSAll the grafted wounds healed very well without any suppuration, bleeding or infection under the grafted skin. No obvious immune rejection was seen. The artificial skin could cover the wounds for a long time with good elasticity and easy to be manipulation.

CONCLUSIONThe composite chitosan artificial skin could be an optimal biological dressing with good histocompatibility and easy to be manipulation.

Animals ; Burns ; surgery ; Cells, Cultured ; Chitin ; analogs & derivatives ; metabolism ; Chitosan ; Dermatologic Surgical Procedures ; Humans ; Keratinocytes ; cytology ; metabolism ; Male ; Rabbits ; Skin ; injuries ; Skin Transplantation ; methods ; Skin, Artificial ; Transplantation, Heterologous ; Wound Healing