Objective To observe biological character iistcs of cultivated cornea limbal epithelial cells of rabbit.Methods Cultivated cornea limbal epithelial cells with tissue inoculation were observed by inverted microscope every two days; ultrastructure of cell was observated by electron microscope; Proliferative ability of cell was examined by flow cytometry; the cell growth curve was examined by monotetrazolium(MTT) colorimetry.Results Cultivated limbal cornea epithelial cells formed monolayer at six day,the cultivated conea limbal epithelial cells had high metabolism and proliferative ability, growth of cornea limbal epithelial cells reached climax after 4 days by passage culture. Conclusion Cultivated limbal epithelial cells during the first and second growth generation can keep cornea epitheliam property and high proliferative ability.

A Ni-based rare-earth perovskite LaNiTiO3 nanoparticles was synthesized and its catalytic activity was investigated. Based on this, a simple and quick nonenzyme electrochemical sensor was fabricated with stable and reliable performances for the determination of hydrogen peroxide (H2 O2 ). The techniques of X-ray diffraction, FT-IR spectra, transmission electron microscopy, X-ray fluorescene spectroscopy and scan electronmicroscope were used to characterize the composition, structure and morphology of as-synthesized sample. The sensor based on this nanomaterial was investigated and optimized by cyclic voltammetry and current-time techniques. The results showed the working electrode modified with LaNiTiO3 (0. 5 g / L, 8. 0μL) in 0. 1 mol/ L NaOH exhibited good catalytic properties for H2 O2 . Under the optimum conditions, the sensor performed excellent properties, such as quick response time ( about 2 s ), a wide linearity (0. 2 μmol/ L -8. 0 mmol/ L), a low detection limit of 0. 05 μmol/ L ( S / N = 3 ), a high sensitivity of 957 μA (mmol/ L) -1 cm-2 , good reproducibility and anti-interference ability, which was better than those of some other biosensors reported recently. So, it may be used for the analysis and detection of H2 O2 in practical samples such as biomedicine.

Radiation-induced lung injury (RILI) is a common complication after radiotherapy for lung cancer and alternative thoracic malignant tumors, while ferroptosis is a regulated cell death triggered by iron-dependent membrane lipid peroxidation. In this article, the relationship between RILI and ferroptosis was investigated from oxidative damage induced by reactive oxygen species, antioxidant network and iron homeostasis regulated by nuclear factor erythroid 2-related factor 2(Nrf2) as well as transforming growth factor involved in the inflammatory response, aiming to mitigate or inhibit the occurrence of RILI through regulating ferroptosis, thereby improving clinical prognosis of patients undergoing radiotherapy.

The present study aims to investigate the kinetic histocytochemical changes of acupoints in different condition. The expression of tryptase (+) mast cells, histamine (HA) , serotonin (5-HT) and nociceptive neuropeptides including calcitonin gene-related peptide (CGRP) and substance P (SP) were observed by immunohistochemistry combined with confocal technology. Mast cells were labeled with anti-mast cell tryptase antibody and simultaneously with HA or 5-HT primary antibodies to observe their co-expression. The results showed that: (1) SP and CGRP were expressed more highly on the cutaneous nerve fibers of "Hegu" (LI 4) after acupuncture stimulation than that of the control. Mast cells aggregated in close proximity to the blood vessels in intra-epidermis and dermis, and some of them with degranulation in the lower dermis and subcutaneous tissue of "Hegu" (LI 4). Both mast cells and their granules appeared with HA (+) and 5-HT (+) expression at stimulated LI 4 sites, while a few intact mast cells with a little expression of 5-HT and HA were distributed in areas of non-stimulated Ll 4. (2) The acupoints in different locations such as Baihui (GV 20), Weishu (BL 21), Zhongwan (CV 12) and LI 4 had the same constituent but the contents were different. (3) The histocytochemical responses of acupoints sensitized by the Gastric mucosa injury (GMI) were also investigated. GMI resulted in neurogenic plasma extravasation by Evans Blue (EB) in the skin of the acupoints over the back and abdomen, which mostly occurred in the T9-T11 dermatomere. The EB extravasation dots just like acupoints sensitization appeared after GMI and disappeared gradually during the natural self-recovery of the gastric mucosa. More SP and CGRP positive nerve fibers were distributed in EB dots than in regions beside EB dots and in the control, mostly distributed in the nerve fibers around both the vessels and root of hair follicle. Mast cells also aggregated and degranulated to release algogenic substances of 5-HT and HA around the vessels in areas of the EB dots. Collectively the acupoints displayed the same histocytochemical responses due to either acupuncture stimulation or GMI. This may potentially be the histocytochemical basis in the local acupoints and acupoints displayed kinetic changes in different condition.
Acupuncture Points ; Acupuncture Therapy ; Animals ; Calcitonin Gene-Related Peptide ; metabolism ; Gastric Mucosa ; chemistry ; metabolism ; Histocytochemistry ; Humans ; Mast Cells ; chemistry ; metabolism ; Mice ; Rats ; Serotonin ; metabolism ; Skin ; chemistry ; metabolism ; Substance P ; metabolism

Objective To investigate the effect of extracellular signal-regulated kinases(ERK) signal transduction in the process of NSCs differentiating into neurons in the fimbria-transected hippocampi's extracts. Methods Twelve Sprague-Dawley rats'right fimbrias were transected. The extracts were gained from the fimbria-transected hippocampi at the 14th day normal rat, and the extracts supernatant fluid was collected after centrifugal process, then the protein concentration in the extracts was determined. In the serum-free medium,NSCs from the fetal hippocampus were planted on 24 well culture plate, then were divided into three group and eight wells for each group as follows: the transected group contained the extracts of the fimbria-transected hippocampi;the normal group contained the extracts of the normal hippocampi;the pure control group have no extracts. After cultured for 14 days,the cells were detected by using MAP-2 and p-ERK immunofluorescence. Result The number, area, perimeter of MAP-2 positive neurons were all declined in transected group, the normal group and the control group orderly. Statistic results showed significant difference between every two groups. The number of MAP-2/p-ERK double-positive neurons were decreased in transected group, the normal group and the control group orderly, but the percentage of double-labeled neurons in total MAP-2 positive neurons were increased in turn. In these two aspect, there were also significant difference between every two group. And most of the MAP-2/p-ERK double-positive neurons were immature. Conclusion The extracts of the fimbria-transected hippocampi had obvious effects on promoting NSCs differentiating into neurons and speeding up the maturation of neurons than those of the normal hippocampi. The morphological results showed that ERK signal transduction might be related to the differentiation of NSCs into neurons.

BACKGROUND: Salvia miltiorrhiza is widely used to treat angina cordis, ischemic stroke and other ischemic cardiovascular diseases. However, the effects of Salvia miltiorrhiza on ovariectomized rats remain unclear.OBJECTIVE: To observe the effects of Salvia miltiorrhiza on the body mass, food intake, and levels of blood lipids and malondialdehyde (MDA) in ovariectomized rats.DESIGN: A completely randomized and controlled experiment.SETTING: Institute of Physiology and Psychology, School of Basic Medical Sciences, Lanzhou University.MATERIALS: The experiment was performed in the Key Laboratory of Pre-clinical Study for New Drugs of Gansu Province and the laboratory of Institute of Physiology and Psychology, School of Basic Medical Sciences, Lanzhou University from November 2005 to December 2006. Twenty-four healthy female SD rats of 3 months old and (220±2) g were selected. Salvia miltiorrhiza water decoction (equal to 1 g/mL crude drug) was identified and extracted by Drug Control Institute of Gansu Province; MDA kit was purchased from Nanjing Jiancheng Institute of Bioengineering.METHODS: ①The rats were randomly divided into three groups with 8 rats in each group: sham-operated group,ovariectomized group and Salvia miltiorrhiza group. The rats were underwent a bilateral ovariectomy except those in the sham-operated group, which were subjected to a removal of bilateral fat as much as ovariectomized group with the ovaries remained. Rats in sham-operated group and ovariectomized group freely drank water; rats in Salvia miltiorrhiza group freely took 1% water extracts from Salvia miltiorrhiza postoperatively, and the concentration of Salvia miltiorrhiza gradually increased to 12% on the eighth day, which was lasted until the end of the experiment (55 days). ②The food intake of rats in each group was monitored daily, and the body mass was measured every five days. At the end of the experiment, femoral artery blood samples of rats were collected to determine the levels of blood lipids. At the same time,MDA was measured according to the kit.MAIN OUTCOME MEASURES: The body mass, food intake, levels of blood lipids and malondialdehyde in each group.RESULTS: Twenty-four rats all entered the result analysis. ①The body mass of rats in 3 groups was nearly the same before operation (P ＞ 0.05). While the body mass in ovariectomized group on the postoperatively 10th, 20th, 25th, and 55th days was significantly higher than those in sham-operated group (P ＜ 0.01). The body mass in Salvia miltiorrhiza group on the postoperatively 20th, 25th, and 55th days was significantly lower than those in ovariectomized group (P ＜0.05-0.01). ②The food intake in ovariectomized group on the postoperatively 15th, 40th, and 55th days was significantly higher than those in sham-operated group (P ＜ 0.05-0.01), and that in Salvia miltiorrhiza group was significantly lower than those in ovariectomized group at those 3 time points (P ＜ 0.05-0.01). ③At the end of the experiment, the levels of total cholesterol, low density lipoprotein cholesterol, high density lipoprotein cholesterol and triglyceride in ovariectomized group were significantly higher than those in sham-operated group (P ＜ 0.05-0.01). The levels of triglyceride and MDA in Salvia miltiorrhiza group were significantly lower than those in ovariectomized group (P＜0.01, 0.05).CONCLUSION: Salvia miltiorrhiza can significantly reduce the body mass and levels of triglyceride and MDA in ovariectomized rats.

Objective To study the apoptotic induction of Aspisol on melanoma A375 cells.Methods The growth inhibition effect of Aspisol on A375 cells was evaluated by MTY assay.Morphological assay was observed by inverted microscope.Early apoptofic rate in A375 cells Was anyzed by flow cytometry.Results The results of MTT assay demonstrated that the growth of A375 cells was inhibited by Aspisol in a time-and concentration-dependence (P＜0.01).FCM analysis showed that Aspisol could markedly induce apeptosis;the early apoptotie rate of A375 cells exposed to different concentrations of Aspisol for 24 h were significant differences(P＜0.05)compared with control group.Conclusion Aspisol could inhibit proliferation and induce apoptosis of A375 cells.

OBJECTIVE To explore the safe and effective dose of sirolimus (Rapamycin,Sir) and its effect on seizure comorbidities. METHODS Immature C57BL/6 mice at postnatal 10 d of age were administered with kainic acid(KA) 12.0 mg · kg-1 intraperitoneally by a single injection to induce acute seizure. Sir 0.3, 1.0 and 3.0 mg · kg-1 was injected 24 h after seizure every other day until 3 d, 1 week, 3 weeks, 5 weeks and 6 weeks. Western blotting analysis was used to detect the expression and phos?phorylation level of S6 protein and to determine the minimum effective dose of Sir. Effect of the mini?mum effective dose of Sir on cognitive function and body growth was observed by several evaluations. Immunofluorescent intensity of Doublecortin (DCX) immunofluorescent staining was conducted to evaluate the development of neurons in the hippocampus. Morris water maze was used to assess the cognitive function. Tail suspension test, O maze and new object recognition test were used to study the anxiety-like behaviors of mice. RESULTS The result of Western blotting showed that Sir 0.3 mg · kg-1 had no significant effect on the phosphorylation of S6 protein in normal mice or KA mice, whereas 1.0 and 3.0 mg · kg- 1 could significantly inhibit the phosphorylation of S6 protein in KA mice (P<0.05). Sir 1.0 mg·kg-1 had no obvious effect on DCX-positive cells or body wass. Morris water maze showed that KA-induced seizure resulted in prolonged escape latency and swimming length (P<0.05), and a decreased crossing number of target quadrant (P<0.05). Sir 1.0 mg·kg-1 significantly reversed the deficit of cognitive function of KA-induced seizure mice (P<0.05), whereas no significant difference was found between Sir group and normal control group. Compared with normal control group, model group showed increased freezing time in tail suspension test (P<0.05), decreased migration length and reten?tion time in open arms in O maze (P<0.05), decreased retention time and touch frequency with new objects, migration length and average speed in new object recognition test (P<0.05). Sir 1.0 mg · kg-1 significantly reversed the above anxiety and depression status, whereas no significant difference was found between sirolimus group and normal control group. CONCLUSION Sir 1.0 mg · kg-1 inhibits the abnormal activation of mTOR pathway and the formation of epilepsy comorbidity in immature mice. Along with its mild side effect in development, Sir 1.0 mg · kg-1 will be an ideal dose to be used in the treatment of seizure in immature mice.

AIM: To investigate multi-potential of rat bo ne marrow mesenchymal stem cells (rBMMSC) and mutation inclination, the rBMMSC w ere long passaged in vitro. METHODS: Cellular cycles of diff erent passages were assayed by FA CSan flow cytometry and karyotypes of passage 6, passage 25 and passage 45 were compared by G-binding analysis. RESULTS: The early passages and long-term passages all showed st rong proliferation; passage 6, passage 25 and passage 45 all showed normal karyo type. CONCLUSION: Long-term culture and passage of rBMMSC still remain s strong proliferation. With this capability, the mutation inclination is not enhanced.