Objective To investigate the effect of leptin on the tolerance of cultured rat brain astrocytes to ischemia and hypoxia.Methods The brain astrocytes isolated from neonatal SD rats,after purification and identification,were incubated in serum-and glucose-flee medium in the presence of 5%CO2+95%N2 for 90 min to induce isehemic and hypoxic injury. RT-PCR was performed to detect the expressions of the leptin receptors Ob-Ra and Ob-Rb in the cells, and colorimetry was used to measure the content of malonaldehyde(MDA) and lactate dehydrogenase(LDH) activity in the cell supematant.The expression level ofglial fibrillary acidicprotein(GFAP)in the cells was detected with fluorescence immunocytochemistry.Results Ischemic and hypoxic exposure of the cells induced obvious cell necrosis.Compared with the cells without the exposure,significantly decreased Ob-Rb expression(0.52±0.01 vs 1.32±0.01,P<0.05)and increased MDA,LDH and GFAP levels(709.68±47.16 vs 516.13±29.08,3.94±0.36 vs 1.81±0.21,and 0.122±0.016 vs 0.057±0.006,respectively,P<0.05) occurred after the exposure,whereas the expression level of Ob-Ra underwent no significant changes(3.87±0.13 vs 3.96±0.24,P>0.05). Compared with the exposed cells,the leptin-treated cells showed a significant reduction in MDA levels(3.94±0.36 vs 3.19±0.25,P<0.05) with significantly increased GFAP expression(0.057±0.006 vs 0.109±0.008, P<0.05)after the exposure, and the cells maintained basically intact cell morphology.Conclusion With neuroprotective effects against ischemic neuronal injuries,leptin canimprove the tolerance of rat brain astrocytes to ischemia and hypoxia.

OBJECTIVETo study the effect of dihydroartemisinin (DHA) combined irradiation on the apoptosis of human lung cancer GLC-82 cells and to study its mechanism.

METHODSThe growth inhibition rate of GLC-82 cells acted by different concentrations DHA was detected using MTT assay at 24, 48, and 72 h, respectively. Clone forming test was used. With multi-target single-hit model, the radiosensitization effect was assessed by calculating sensitizing enhancement ratio (SER).The effect of DHA combined irradiation on the apoptosis of GLC-82 cell cycle distribution and apoptosis were measured by flow cytometry. The protein expression of p53, p21, Bcl-2, and Bax were detected by Western blot.

RESULTSDifferent concentrations DHA (4, 8, 16, 32, 64, and 128 μg/mL) had cytotoxicity on GLC-82 cells. The IC50 for 24, 48, and 72 h was 38.25,20.58, and 10.36 μg/mL, respectively, in obvious dose- and time-dependent manner. The growth inhibition rate was more significantly increased than that of the blank control group (P < 0.01, P<0.05). DHA had sensitization enhancement effect on GLC-82 cells, with SER of 1.4. DHA combined irradiation could obviously change the structure of GLC-82 cells cell cycle and induce apoptosis (with the apoptosis rate of 21.5%), which was significantly different from that of the blank control group (P < 0.05). Western blot showed the expression of p53 and p21 protein could be increased by DHA combined irradiation, and the expression of Bcl-2 protein down-regulated (P <0.01, P <0. 05).

CONCLUSIONSDHA had stronger cytotoxicity and radiosensitization on GLC-82 cells. Its mechanisms might lie in making the arrest of GLC-82 cells' growth at G0/G1 phase, decreasing the ratio of cells at S phase, restoring the function of p53, decreasing the expression of Bcl-2 protein, and inducing apoptosis in GLC-82 cells.

Apoptosis ; drug effects ; Artemisinins ; pharmacology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Down-Regulation ; drug effects ; Flow Cytometry ; Humans ; Lung Neoplasms ; metabolism ; Neoplasm Proteins ; metabolism ; Radiation-Sensitizing Agents ; pharmacology ; Tumor Cells, Cultured ; bcl-2-Associated X Protein ; metabolism

Objective To study the effects of Jianpi Xiaoai Prescription on cell cycle and apoptosis of human colon cancer HCT116 cells and related factors of Wnt/β-catenin signaling pathway; To investigate its mechanisms of anti-metastasis and recurrence of colorectal cancer. Methods The logarithmic growth phase HCT116 cells were divided into blank group, saline group, 5-Fu group, and Jianpi Xiaoai Prescription low-, medium-, and high-dose groups. After intervention for 48 h, the cells were harvested, and the cell cycle and apoptosis were detected by flow cytometry. The protein expression of β-catenin in the nucleus was detected by Western blot, and the expression of c-myc and cyclinD1 mRNA was detected by PCR. Results Compared with the blank group and saline group, the ratio of HCT116 cells apoptosis of Jianpi Xiaoai Prescription low-, medium-, and high-dose groups increased; the proportion of cells in phase G1 increased and the proportion of S cells decreased; the expression of β-catenin protein in the nucleus and the expression of c-myc,cyclinD1 mRNA decreased,especially in the Jianpi Xiaoai Prescription high-dose group,with statistical significance(P<0.05,P<0.01).Conclusion Jianpi Xiaoai Prescription can promote apoptosis of human colon cancer HCT116 cells and block the cell cycle, and its mechanism may be related to regulating Wnt/β-catenin signaling pathway.

PURPOSE: Previous western studies have found Caucasians with skin cancer, either melanoma or nonmelanoma skin cancer (NMSC), have an elevated risk of second primary cancer. Our objective was to assess the risk of second primary cancer in Taiwanese with NMSC. MATERIALS AND METHODS: By using data from Taiwan's National Health Insurance Research Database, we conducted a population-based cohort study to assess the risk of incident second primary cancer in Taiwanese affected by NMSC. RESULTS: We identified 505 subjects with NMSC and 2,020 matched controls. After adjustment for potential confounders including age, sex, urbanization, and Charlson Comorbidity Index, people who had NMSC had a 1.43-fold (95% confidence interval [CI], 1.05 to 1.96) risk for the development of second primary cancer as comparedwith control group. Menwith NMSC had a 2.99-fold (95% CI, 1.00 to 9.10) risk for second primary cancer involving the lip, oral cavity, and pharynx and a 3.51-fold (95% CI, 1.21 to 10.17) risk for second primary cancer involving the genitourinary organs when compared to the control group. By contrast, women with NMSC did not have an increased risk of second primary cancer. CONCLUSION: This study revealed Asians with NMSC have an increased risk of second primary cancer. Our findings can be a useful reference for health care for people diagnosed with NMSC.
Asian Continental Ancestry Group ; Cohort Studies* ; Comorbidity ; Delivery of Health Care ; Female ; Humans ; Lip ; Melanoma ; Mouth ; National Health Programs ; Neoplasms, Second Primary* ; Pharynx ; Skin Neoplasms* ; Skin* ; Urbanization

Virtopsy is a non-invasive technique to reconstruct 3 dimensional (3-D) images of human organs and tissues using digitized radiographic imaging and may provide clues for forensic identification of the cause and manner of death. Because of its nature of minimally invasive, objective, and accurate, virtopsy has recently been a research focus of forensic pathology in developed countries. In this review article, the authors will discuss the principle, advantage, disadvantage, and recent proceeding of virtopsy as well as its potential application in forensic practice in China.
Autopsy ; Forensic Medicine/methods* ; Forensic Pathology ; Humans ; Imaging, Three-Dimensional/methods* ; Magnetic Resonance Imaging/methods* ; Tomography, X-Ray Computed/methods*

OBJECTIVETo explore the electrophysiological properties of differentiation of rat bone marrow-derived stromal stem cells (rBMSCs) to neuron-like cells in vitro by edaravone, a new type of free radical scavenger.

METHODSStromal stem cells were separated from rat bone marrow with Ficoll-Paque reagent and expanded in different culture medium in vitro. rBMSCs were induced by edaravone containing serum-free L-DMEM. Morphologic observation and Western blot analysis including the expression of Nav1.6, Kv1.2, Kv1.3, Cav1.2 were performed, and whole patch-clamp technique was used.

RESULTSCyton contraction and long processes were shown in differentiated stromal stem cells. Nav1.6, Kv1.2, Kv1.3 and Cav1.2 were expressed in both differentiated and undifferentiated cells. However, the expression of channel proteins in differentiated cells was up-regulated. Consistently, their resting potential and outward currents were also enhanced in the differentiated cells, which was especially significant in the outward rectifier potassium current.

CONCLUSIONIn vitro, neuron-like cells derived from rBMSCs, induced by edaravone, possess electrophysiological properties of neurons.

Animals ; Antipyrine ; analogs & derivatives ; pharmacology ; Blotting, Western ; Bone Marrow Cells ; cytology ; physiology ; Cell Differentiation ; drug effects ; Male ; Neurons ; cytology ; physiology ; Rats ; Rats, Sprague-Dawley ; Stromal Cells ; cytology ; physiology

OBJECTIVETo compare the efficacy differences of electroacupuncture (EA) with intermittent wave, disperse-dense wave and continuous wave combined with rehabilitation therapy for post-stroke shoulder subluxation in order to provide clinical evidence for selection of electroacupuncture wave.

METHODSSeventy-five cases of shoulder subluxation were randomly divided into an intermittent wave (IW) group, a disperse-dense wave (DW) group and a continuous wave (CW) group, 25 cases in each one. Based on regular treatment including blood pressure and sugar control, EA and rehabilitation therapy were given in three groups. With EA at Jianqian (Extra), Taijian(Extra), Binao (LI 14), Jianzhong (Extra), Jianzhen (SI 9), Naoshu (SI 10), Jianwaishu (SI 14) and Bingfeng (SI 12), IW group was treated with intermittent wave with a frequency of 15 Hz, DW group was treated with disperse-dense wave with a frequency of 2 Hz/15 Hz and CW group was treated with continuous wave with a frequency of 15 Hz. The EA treatment was given for 20 min, once a day, six times a week and totally for 4 weeks. The Fugl-Meyer score and Barthel index score were used for assessment of function recovery before and after treatment respectively, also the efficacy of three groups was compared.

RESULTSThe total effective rate was 96.0% (24/25) in the IW group, which was superior to 68.0% (17/ 25) in the DW group and 64.0% (16/25) in the CW group (both P < 0.05). After the treatment, Fugl-Meyer score and Barthel index score in the three groups were obviously improved (all P < 0.05), which was most significant in the IW group (all P < 0.01).

CONCLUSIONElectroacupuncture with intermittent wave combined with rehabilitation therapy has superior effect to disperse-dense wave and continuous wave on the treatment of post-stroke shoulder subluxation, which could effectively improve shoulder subluxation and promote shoulder function recovery.

Aged ; Combined Modality Therapy ; Electroacupuncture ; Female ; Humans ; Male ; Middle Aged ; Shoulder ; physiopathology ; Shoulder Dislocation ; etiology ; rehabilitation ; therapy ; Stroke ; complications ; Treatment Outcome

OBJECTIVETo study the feasibility of MRI of human colon adenocarcinoma cell line (Lovo) labeled with superparamagnetic iron oxide(SPIO) nanoparticles in vitro.

METHODSLovo cells (5 × 10(5) and 1 × 10(6)) were cultured in medium containing different SPIO nanoparticles (50 microl and 500 microl). Transmission electron microscopy was used to observe cellular ultrastructure and to determine the uptake and distribution of particles in Lovo cells at 1-, 3-, 6-hours. MRI of Lovo cells was performed with T1WI, T2WI sequences. Unlabeled cells were used as controls.

RESULTSUptake of SPIO nanoparticles occurred within 6 hours. On T1 weighted imaging, there was no significant difference in signal intensity between the experimental groups and the control group. On T2 weighted imaging, there was no significant difference in signal intensity between the experimental groups and the control group after culture of 1 h. Signal intensity began to decrease in 1 × 10(6) Lovo cells labeled with 500 microl SPIO nanoparticle after 3 hours culture. Signal intensity decreased in all the experimental groups after 6 hours culture.

CONCLUSIONHuman colon adenocarcinoma cell line (Lovo) can be labeled with SPIO nanoparticles, and the labeled cells can be imaged with MRI equipment.

Adenocarcinoma ; pathology ; Cell Line, Tumor ; Colonic Neoplasms ; pathology ; Humans ; Iron ; Magnetic Resonance Imaging ; methods ; Magnetics ; Nanoparticles ; Oxides ; Pilot Projects ; Staining and Labeling ; methods

OBJECTIVETo establish a new method for the preparation of porcine acellular dermal matrix.

METHODSThe antigenicity of the porcine dermis was weakened by removing epidermal and dermal cells from the porcine skin through the digestion with low-concentration trypsin and repeated freeze-thaw cycles. Split thickness porcine skin was treated with 0.05% trypsin to remove the cells from the epidermis and dermis. Repeated freeze-thaw cycles were employed to further weed out the residual cells within the dermis. The prepared acellular dermis was then examined grossly, as well as histologically, and also by immunohistochemical method.

RESULTSNo cell could be identified in the prepared porcine acellular dermal matrix. The integral basement membrane was preserved on the surface of dermal matrix with compact dermal matrix collagen structure.

CONCLUSIONLow concentration trypsinization and repeated freeze-thaw cycles seemed to be a simple and effective method for the preparation of xenogeneic acellular dermal matrix.

Animals ; Dermis ; cytology ; transplantation ; Extracellular Matrix ; transplantation ; ultrastructure ; Freezing ; Skin Transplantation ; Swine ; Tissue Engineering ; methods ; Trypsin ; administration & dosage

Aim To observe the absorption activity of five components of Polygonum orientale L. Flower ex-tract in Caco-2 cell monolayer. Methods The effects of different concentrations, time, temperature, pH and P-glycoprotein inhibitors on Caco-2 cells were investi-gated by UPLC-MS/MS. Results The absorption of five components of Polygonum orientale L. Flower ex-tract presented a concentration-and time-dependent manner, and the uptake of quercetin was reduced in Caco-2 cells after 90 min. There was a highest intake at 37℃,and the uptake of four components was best in acid environment except for the quercetin at pH 6 , which was best at pH5 . The uptake of quercetin and kaempferol was significantly improved after the addition of P-glycoprotein inhibitors of verapamil. Conclusions The cellular uptake mechanisms of the five compo-nents of Polygonum orientale L. Flower extract is man-inly through passive diffusion. P-glycoprotein is in-volved in the uptake of quercetin and kaempferol.