Autophagy is a crucial biological process of eukaryotes, which is involved in cell growth, survival and energy metabolism, while the premise of the autophagy function is activated autophagic flux. It has been confirmed that impaired autophagic flux promotes pathogenesis of many chronic inflammatory diseases, especially cancer, neurodegenerative disease and tissue fibrosis, therefore the analysis of autophagic flux state is important for revealing autophagy function and the mechanism of autophagy related diseases. Given that autophagy is a dynamic process with multiple steps, it is very hard to observe the real state of autophagic flux. Summarized here is the novel concept and current approach to detect autophagic flux. This knowledge is crucial for the researching of the biological function of autophagy, and may provide some strategies for developing autophagy-related drug.
Autophagy ; Fibrosis ; Humans ; Inflammation ; pathology ; Neoplasms ; pathology ; Neurodegenerative Diseases ; pathology

BACKGROUND: It is significant to establish a kind of effective, conve nient and reliable animal model of hypertension. At present, dogs, rats and rabbits are usually used to establish hypertensive models at home and abroad, and the renal artery stenosis induced hypertensive models are ex tensively used to research hypertension and its complication for human be ings because they are convenient and reliable, and there are many methods to establish them, but the effects are to be evaluated. OBJECTIVE: To establish convenient and reliable animal models of ex perimental renal artery stenosis induced hypertension. DESIGN: A randomized grouping design and animal experiment. SETTING: Institute of Cerebrovascular diseases, Medical College Hospital of Qingdao University. MATERIALS: The experiments were carried out in Shandong Key Labora tory for Prevention and treatment of Brain Disease from September 2005 to February 2006. Eighty-one healthy Wistar rats divided into 7 groups accord ing to the method of random number table: unilateral renal artery stenosis group (n=18), bilateral renal artery stenosis group (n=17), unilateral renal artery ligation group (n=15), bilateral renal artery ligation group (n=15), uni lateral renal artery stenosis sham-operated group (n=6), bilateral renal artery stenosis sham-operated group (n=4) and normal control group (n=6). METHODS: Unilateral renal artery stenosis group: Right renal artery was clamped with miniature silver clip, and left kidney was resected after 12 days. Bilateral renal artery stenosis group: Right renal artery was clamped with miniature silver clip, and the same treatment was given to the left side after 12 days. Unilateral renal artery ligation group: Right renal artery was ligated with filament, and left kidney was resected after 12 days. Bilateral renal artery ligation group: Right renal artery was ligated with filament, and the same treatment was given to the left side after 12 days. Unilateral renal artery stenosis sham-operated group: Right kidney was exposed, and returned to the original place without any treatment, and left kidney was resected af ter 12 days. Bilateral renal artery stenosis sham-operated group: Right kid ney was exposed, and returned to the original place without any treatment, and the same treatment was given to the left side after 12 days. Normal con trol group: The rats were not given any treatment. The blood pressure and heart rate were determined with RBP-2 hemomanometer for rats. MAIN OUTCOME MEASURES: The successful rate of model estab lishment, blood pressure and heart rate were observed. RESULTS: Totally 81 rats were used, and 61 of them died, all were in volved in the analysis of results without deletion. ① Blood pressures in the unilateral and bilateral renal artery stenosis groups and bilateral renal artery ligation group were obviously higher than those in the normal control group and bilateral renal artery stenosis sham-operated group [(138.0 ±36.5), (154.2±11.6), (160.5±0.7), (101.3±17.6), (108.3±5.7) mm Hg]. ② The changes of heart rate in the renal artery stenosis group were unstable, and the heart rates in the unilateral and bilateral renal artery stenosis groups, bilateral renal artery ligation group, normal control group and bilat eral renal artery stenosis sham-operated group were (367.5±47.2), (420.2 ±47.8), (386.0±4.2), (390.3±42.4), (417.3±27.5) beats per minute, respec tively. ③ The survival rates in the renal artery stenosis groups (22%, 29%) were significantly higher than those in the renal artery ligation groups (0,12%), and it was the highest in the unilateral renal artery stenosis group.CONCLUSION: The method of clamping bilateral renal arteries can establish stable rat models of hypertension induced by renal artery stenosis.

Incidence of cancer is increasing year by year,diagnostic techniques are continually advanced,and the numbers of anti-cancer drugs are growing,but the efficacy has not been fundamentally improved.Surgery,chemotherapy and radiotherapy can only kill the part of mature tumor cells,but can not kill the cancer stem cells,and eventually leading to tumor recurrence.While acupuncture,qigong,psychology comprehensive treatment can inhibit tumor growth and reduce metastasis and recurrence,improve quality of life and prolong survival.We should update our recognition,to carry out the study on the treatment of tumor by TCM with large sample and multiple centers,to demonstrate its low toxicity,low costs and efficiency.

BACKGROUND: A proper preservation method would be of important significance for experiments and clinical application ofolfactory ensheathing cells (OECs) OBJECTIVE: To explore proper cyropreservative systems for OECs.METHODS: OECs during the logarithmic growth phase were harvested, cryopreserved for 1, 3 and 6 months and then revitalized.RESULTS AND CONCLUSION: MTT assay and tryplan blue staining showed that cells exhibited highest viability after treatmentwith 5% dimethyl sulfoxide (DMSO)-6% hydroxyethyl starch (HES), followed by 10% DMSO, and lastly the 5% DMSO. Use ofrefrigerator or cryogenic control system with different cryopreservation time did not yield obvious effects on viability of OECs.Therefore, 5% DMSO-6%HES is recommended as a cryopreservative agent for OECs.

AIM:To observe the effects of the combination of berberin and epirubicin on the cell cycle of T24 bladder cancer cells and the underlying mechanisms.METHODS:The cancer cells were exposed to epirubicin in the presence or absence of different concentrations of berberin.The viability of the cancer cells was determined by MTT assay.The cell cycle distribution was detected by flow cytometry, and the protein levels of cyclin D1, CDK2, CDK4, P21 and P27 were detected by Western blot.RESULTS:Berberine markedly enhanced the inhibitory effect of epirubicin on the viability of T24 cells and promoted epirubicin-induced cell cycle arrest at G0/G1 phase as compared with the negative control cells.Epirubicin increased the protein expression of P27 and P21, both of which were enhanced by treatment with berberin.In contrast, berberin exposure further decreased the protein expression of cyclin D1, CDK2 and CDK4 in epirubicin-treated T24 cells.CONCLUSION:Berberine significantly promotes epirubicin-induced G0 /G1 phase arrest in human bladder cancer cells by up-regulating P27 and P21 expression and inhibiting the expression of cyclin D1, CDK2 and CDK4.

Chemokines are small cytokines with chemotactic activity, they are involved in regulating immune responses and inflammatory responses. In the development of tumors, chemokines are multi-functional mediators that not only affect the infiltration of immune cells into the tumor, but also have an important impact on tumor growth, angiogenesis, invasion, and metastasis. Besides, they are important targets of tumor therapy. Here we review chemokines involved in the regulation of signaling pathways, analyze the mechanism of chemokines in the development of breast cancer, summarize the chemokines targeted drugs for breast cancer in recent years and make a prospect about the role of chemokines in anti-breast cancer therapy.

Endogenous ouabain (EO) is a recently found hormone that may be secreted from adrenal cortex. As an endogenous mammalian analogue of cardiac glycosides and an inhibitor of the sodium pump, it regulates the body fluid balance, urine sodium extraction and vasoconstrictive tone, and thus plays an important role in the pathogenesis of hypertension and some other cardiovascular disorders. This articke reviews its biological features, receptor and antibody, detection, effects on diseases and relationship with endothelial cells.
Adrenal Cortex ; metabolism ; Animals ; Antibodies ; therapeutic use ; Humans ; Hypertension ; etiology ; Nephrotic Syndrome ; etiology ; Ouabain ; immunology ; pharmacology ; Sodium-Potassium-Exchanging ATPase ; immunology ; physiology

Objective To investigate the clinical results of the primary total hip replacement (THR) and the secondary THR after failed internal fixation in the elderly patients with displaced femoral neck fracture so as to find the optimal treatment for displaced femoral neck fractures in the elderly patients. Methods From April 2001 to April 2007,16 patients (Study Group) treated with a secondary THR after failed internal fixation and 20 patients (Control Group) treated with a primary THR were enrolled in the study and followed up. There were seven males and nine females, at average age of 66. 5 years (50-85 years) and with mean follow-up period of 58. 25 months (24-96 months) in the Study Group. There were six males and 14 females, at average age of 68.1 years (51-83 years) and with mean follow-up period of 49.50 months (24-70 months) in the Control Group. All patients were active and lucid before they suffered fractures. Blood loss and operation duration in THR were compared. Hip function (Harris score) and health-related quality of life (HRQoL, KPS index score) were assessed during the follow-up after THR. Results Operative duration was (115.63 ±34.35) minutes in Study Group and (91.25 ±15.80) minutes in Control Group (P<0.05). Blood loss was (546.86 ±377.04) ml in Study Group and (320.00 ±155.94) ml in Control Group (P<0.05). At follow up, Harris score and KPS index score were (87. 25 ±7. 53) points and (95. 00 ±5. 16) points respectively in Study Group, and (90.20±5.46) points and (96.00 ±0.73) points respectively in Control Group (P>0.05). There were no infections or re-operations in two groups, but with one death in each group during the follow-up. Conclusions THR is the optimal treatment for displaced femoral neck fractures in the elderly patients.The secondary THR after failed internal fixation has higher risks in operation compared with the primary THR for a displaced femoral neck fracture in the elderly patient.

Octapeptin has strong antibacterial activity against Gram-negative bacteria such as Escherichia coli, Klebsiella pneumoniae and Acinetobacter baumannii, while it also has activity against some Gram-positive bacteria. This study used natural octapeptin A3 and B3 as lead compounds for structural modification. Twenty-one peptide derivatives (including A3 and B3) containing eight amino acid residues were prepared by solid-phase synthesis, and evaluated for antibacterial activity and renal cytotoxicity. Among them, three compounds 6, 7 and 17 exhibited broad-spectrum antibacterial activity and significantly enhanced the activity for Gram-positive bacteria while maintaining the activity of Gram-negative bacteria. Several compounds improved the activity for Pseudomonas aeruginosa. Compound 7 was active against all test strains and had relatively low renal cytotoxicity. The results provide a basis for the further development of novel polypeptide antibiotics.

OBJECTIVETo investigate the role of renal sympathetic nerves in renal sodium transport in ouabain-hypertensive rats (OHR).

METHODSSixteen male SD rats with sham renal denervation (Sham-RDNX) and 16 with renal denervation (RDNX) were randomly into normal control group and ouabain group to receive intraperitoneal injection of normal saline (1 ml/kg) and ouabain (27.8 µg/kg) once a day, respectively. Systolic blood pressure (SBP), heart rate and body weight were recorded weekly. Food consumption of the rats was determined twice a week. After a 4-week treatment, blood and 24 h urine samples were collected to measure the serum and urinary concentration of sodium, trace lithium and creatinine. Endogenous creatinine clearance rate (Ccr), fractional excretions of sodium (FENa), fractional excretions of lithium (FELi) and fractional reabsorption of sodium in the postproximal tubules (FDRNa) were calculated. Plasma renin activity was determined by radioimmunoassay. Norepinephrine was extracted from the renal tissue and assayed for norepinephrine content by HPLC.

RESULTSThe body weight, food intake and heart rate showed no significant difference among the 4 groups (P > 0.05). After 4 weeks, the SBP of control RDNX group (CDNX) was significantly lower than that of the control Sham-DNX group (Csham)(P < 0.05); the SBP of ouabain RDNX group (ODNX) was also significantly lower than that of ouabain Sham-DNX group (Osham) (P < 0.05); RNDX lowered SBP by about 10 mmHg in both ouabain groups and control groups. The SBP was significantly higher in Osham and ODNX groups than in the corresponding control groups (P < 0.01), also significantly higher in ODNX group than in Csham group (P < 0.01). Ccr showed no significant difference among the 4 groups(P > 0.05). FENa, FELi and FDRNa were significantly lower in ouabain groups than in the corresponding control groups (P < 0.05, P < 0.01, P < 0.05), but FENa, FELi and FDRNa of ODNX group were similar with those of Osham group (P > 0.05); FENa , FELi and FDRNa were similar between CDNX and Csham groups (P > 0.05). The plasma renin activity was comparable between the 4 groups (P > 0.05). Renal norepinephrine level was markedly reduced in RDNX group compared with that in Sham-RDNX group in both ouabain and control groups (P < 0.01).

CONCLUSIONThe increase of proximal tubule sodium reabsorption in OHR is not dependent on the renal sympathetic nerve.

Animals ; Hypertension ; chemically induced ; metabolism ; Kidney ; innervation ; Male ; Ouabain ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sodium ; metabolism ; Sodium Channels ; metabolism ; Sympathetic Nervous System ; physiology