Child ; Digestive System ; metabolism ; Glutamine ; metabolism ; Humans ; Immune System ; metabolism ; Nutritional Physiological Phenomena ; physiology

AIM: To optimize the conditions for in vitro culture of retinal pigment epithelium (RPE) cells, we characterized expressions of various growth factors in RPE cells, including tumor necrosis factor (TNF-α), vascular endothelial growth factor (VEGF), β fibroblast growth factor (βFGF), transforming growth factor β2 (TGFβ2), and interferon-γ (IFN-γ). We also studied expressions of caspase-3 under different concentrations of fetal bovine serum (FBS) with insulin-transferrin-sodium selenite (ITS) supplement. METHODS: First, we investigated if expressions of TNF-α, VEGF, βFGF, TGFβ2, IFN-γ, and caspase-3 in FBS and ITS with of concentration. Second, we cultured primary RPE cells from eyes of forty C57 BL/6 mice in standard dulbecco's modified eagle's medium (DMEM) containing 20,40,100mL/L FBS and 20,40,100mL/L FBS together with 10g/L ITS. Immunohisto-chemical staining and cell counting were performed to verify the existence and growth condition of RPE cells. Expressions of TNF-α, VEGF, βFGF, TGFβ2 and IFN-γ were determined using cells and supernatant from passage-3 to -4 primary RPE cell after 48 hours of culture with RT-PCR and enzyme-linked immunosorbent assays (ELISA). The expression of casepase-3 was determined via Western blotting. The major outcome measurement is the expression level of growth factors in cultured RPE cells and the experiment design is to expose the RPE cells to different culture medium. RESULTS: TNF-α, VEGF, βFGF, TGFβ2, but not IFN-γ, were expressed and the expressions increased with concentration. No expression of the aforementioned genes was detected in presence of ITS. The primary cultures of RPE cells were successfully established. TNF-α, VEGF, βFGF, TGFβ2 (but no IFN-γ) and the active caspase-3 were detected in 20,40,100mL/L FBS or 20,40,100mL/L FBS combined with 10g/L ITS; the expressions were upregulated with increasing concentration of FBS. There is no significant difference in the expression of growth factors between these groups. However, significant differences were shown among different concentration of FBS (P＜0.01）. The lowest expression was observed in 20mL/L FBS or 20mL/L FBS combined with 10g/L ITS medium with RPE cells. But RPE cells were shown in better growth condition in 20mL/L FBS combined with 10g/L ITS.CONCLUSION: TNF-α, VEGF, βFGF, TGFβ2 and caspase-3 were expressed in RPE cells and supernatants. The production of above 20mL/L FBS combined with 10g/L ITS in DMEM may be the ideal cell culture medium that supports the normal growth of RPE cells.

OBJECTIVETo explore effects of Shenqi preparation,Traditional Chinese Medicine, on anti-fatigue and anti-oxidant functions.

METHODSOne hundred and twenty mice were randomly divided into control group and 3 experimental groups. The high, medium and low-dose of Shenqi preparation were given to the 3 experimental groups respectively, while distilled water to the control group for 15 d. The loaded swimming time, the level of lactate, serum urea nitrogen (SUN), muscle and liver glycogen, liver super-oxide dismutase (SOD), the content of malondialdehyde (MDA), glutathione peroxidase(GSH-Px) were assayed.

RESULTSThe loaded swimming test showed that the exhausted swimming time of 3 experimental groups [(296.0 +/- 25.3)s, (437.0 ĝ 38.9)s, (595.0 +/- 53.9)s respectively] was longer than that of control group [(231.0 +/- 22.5)s, P < 0.05, P < 0.01]. The liver glycogen content of the high and medium-dose experimental groups were higher than that of control group respectively (P < 0.01). The SUN content of each experimental group was less than that of the control group (P < 0.01, P < 0.05). Moreover,in the medium and high dose experimental groups, less accumulation of lactate was found (P < 0.01, P < 0.05), and the content of liver SOD and GSH-Px was higher (P < 0.01, P < 0.05). The content of liver MDA in high-dose experimental group was less than that of the control group (P < 0.05).

CONCLUSIONShenqi preparation, especially the high and medium-dose experimental groups, is able to improve exercise tolerance and has anti-fatigue and anti-oxidant effects in mice.

Animals ; Antioxidants ; metabolism ; Blood Urea Nitrogen ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Fatigue ; drug therapy ; metabolism ; Glutathione Peroxidase ; metabolism ; Glycogen ; metabolism ; Lactic Acid ; blood ; Liver ; drug effects ; metabolism ; Male ; Malondialdehyde ; metabolism ; Mice ; Physical Conditioning, Animal ; Superoxide Dismutase ; metabolism

Adolescent ; Adult ; Follow-Up Studies ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Recovery of Function ; Tibial Fractures ; diagnostic imaging ; physiopathology ; surgery ; Tomography, X-Ray Computed ; Young Adult

Insulin receptor substance-1 (IRS-1) mRNA and protein were assayed in rat muscle of hindlamb by RT-PCR and immunohistochemistry respectively.Smoking decreased the expressions of IRS-1 mRNA and protein in rat muscle of normal chow smoking group,high fat chow smoking group and diabetic smoking group as compared with matched control groups (P

Cancer is a group of various diseases, all of which involve unregulated cell growth. Many currently used chemotherapeutic drugs are derived from botanicals. Thus, searching botanical sources for novel oncology medications, including identifying the lead compounds and their derivatives for chemoprevention, is an essential step in advancing cancer therapeutics. This article mainly focuses on the data from our previous American ginseng anti-colon cancer studies. In addition to the potential role of American ginseng on cancer, the herb as an adjuvant for cancer treatment is presented, including describing the attenuation of adverse events induced by chemotherapeutic agents and increasing of quality of cancer patient life. Since heat-treated American ginseng and ginsenoside gut microbiome metabolites showed significant increases in cancer chemopreventive effects, active constituents of the steamed herb and their gut metabolites should be clearly identified, and the structure-activity relationship should be further explored. Data obtained from herbal medicine studies and clinical trials will help develop useful anticancer agents.
Animals ; Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Colorectal Neoplasms ; drug therapy ; pathology ; Ginsenosides ; isolation & purification ; metabolism ; pharmacology ; therapeutic use ; Hot Temperature ; Humans ; Panax ; chemistry ; Phytotherapy ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Structure-Activity Relationship

OBJECTIVETo explore the clinical efficacy of the arthroscopic Mender II stapler for the treatment of patients with meniscus anterior horn injury needing meniscal suture repair.

METHODSAmong 47 patients with meniscus anterior horn injury, 29 patients were male and 18 patients were female, ranging in age from 12 to 31 years old, with a mean age of (20.53± 4.12) years old. The duration of disease ranged from 3 to 35 days, and the average duration was (12.43±5.74) days. The Mender II stapler was used to carry out arthroscopic suture from outside to inside. The Lysholm knee scoring system was used to evaluate and analyze preoperative and postoperative symptoms, such as pain, limping embolism and so on.

RESULTSForty-six patients were followed up, and the duration ranged from 12 to 48 months, with a mean of (20.53±4.12) years. The incision healed at the first stage without important vessels and nerves injuries. The symptoms of the locked knee joint disappeared, and symptoms such as pain, limp, swelling and limitation of activity improved. The Lysholm score increased from preoperative 52.33±7.31 to postoperative 86.74±6.92.

CONCLUSIONUsing Mender II stapler to treat patients with meniscus anterior horn injury who were treated with arthroscopic suture from outside to inside is effective to improve symptoms, and to obtain good short-term results.

Adolescent ; Adult ; Arthroscopy ; methods ; Child ; Female ; Humans ; Male ; Menisci, Tibial ; surgery ; Suture Techniques ; Tibial Meniscus Injuries

Injury or inflammation affecting sensory neurons in the dorsal root ganglia (DRG) causes hyperexcitability of DRG neurons that can lead to spinal central sensitization and neuropathic pain. Recent studies have indicated that, following chronic compression of DRG (CCD) or acute dissociation of DRG (ADD) treatment, both hyperexcitability of neurons in intact DRG and behaviorally expressed hyperalgesia are maintained by activity in cGMP-PKG signaling pathway. Here, we provide evidence supporting the idea that CCD or ADD treatment activates cGMP-PKA signaling pathway in the DRG neurons. The results showed that CCD or ADD results in increase of levels of cGMP concentration and expression of PKG-I mRNA, as well as PKG-I protein in DRG. CCD or ADD treated-DRG neurons become hyperexcitable and exhibit increased responsiveness to the activators of cGMP-PKG pathway, 8-Br-cGMP and Sp-cGMP. Hyperexcitability of the injured neurons is inhibited by cGMP-PKG pathway inhibitors, ODQ and Rp-8-pCPT-cGMPS. In vivo delivery of Rp-8-pCPT-cGMPS into the compressed ganglion within the intervertebral foramen suppresses CCD-induced thermal hyperalgesia. These findings indicate that the in vivo CCD or in vitro ADD treatment can activate the cGMP-PKG signaling pathway, and that continuing activation of cGMP-PKG pathway is required to maintain DRG neuronal hyperexcitability and/or hyperalgesia after these two dissimilar forms of injury-related stress.
Animals ; Cyclic GMP ; analogs & derivatives ; metabolism ; Cyclic GMP-Dependent Protein Kinases ; metabolism ; Ganglia, Spinal ; physiopathology ; Hyperalgesia ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Thionucleotides ; metabolism

OBJECTIVETo further study the role of human cytomegalovirus (HCMV) infection in the pathogenesis of the type 2 diabetes mellitus.

METHODSHCMV DNA levels in sera from 29 type 2 diabetic patients and 23 controls were measured by quantitative polymerase chain reaction (PCR), and comparative analyses was made between HCMV DNA and T cell subsets, blood glucose (BG), insulin (Ins) and C peptide (C-P).

RESULTSThe levels of HCMV DNA were (1.81+/-1.67) x 10(8) copies/ml for type 2 diabetic patients, a level significantly higher than that (5.50+/-4.30) x 10(7) copies/ml of controls. The percentage of CD8 for type 2 diabetic patients with positive HCMV DNA was 29.53%+/-2.00%, being much higher than that for controls (27.13%+/-4.12%), while the ratio of CD4/CD8 1.24+/-0.05 was significantly lower than that 1.41+/-0.10 of controls. Fasting C-P of type 2 diabetic patients with positive HCMV DNA was far lower than that of those with negative HCMV DNA, but the differences of BG and Ins between the two groups were not significant.

CONCLUSIONActive HCMV infection of type 2 diabetic patients may suppress cellular immunity and its influence on the pathogenesis of the type 2 diabetes mellitus should be further studied.

Adult ; Cytomegalovirus ; genetics ; Cytomegalovirus Infections ; complications ; immunology ; DNA, Viral ; blood ; Diabetes Mellitus, Type 2 ; immunology ; virology ; Female ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; T-Lymphocyte Subsets

This paper is to report the study of the pharmacokinetics of a fusion protein TAT-haFGF(14-154) for human acidic fibroblast growth factor and transcriptional activator protein in rat plasma, and the investigation of their penetration across blood-brain barrier in mice and rats, in order to provide a basis for clinical development and treatment of Alzheimer's disease. Enzyme-linked immunosorbent assay (ELISA) was used to determine concentration of TAT-haFGF(14-154) in rat plasma and in mouse brain homogenate; and immunohistochemistry was used to analyze the distribution in brain. The concentration-time curve fitted two-compartment open model which was linear kinetics elimination after a single intravenous injection of TAT-haFGF(14-154) in rat at the dose of 300 microg x kg(-1). The half life time was 0.049 +/- 0.03 h for distribution phase and 0.55 +/- 0.05 h for elimination phase, and the weight was 1/C2. The result showed that TAT-haFGF(14-154) could be detected in the brain by ELISA and immunohistochemistry, the elimination of TAT-haFGF(14-154) in rat was swift, and TAT-haFGF(14-154) could penetrate across the blood-brain barrier, distribute in pallium and hippocampus and locate in the nucleus.
Animals ; Blood-Brain Barrier ; metabolism ; Brain ; metabolism ; Cell Nucleus ; metabolism ; Cerebral Cortex ; metabolism ; Female ; Fibroblast Growth Factor 1 ; administration & dosage ; pharmacokinetics ; Gene Products, tat ; administration & dosage ; pharmacokinetics ; Hippocampus ; metabolism ; Injections, Intravenous ; Male ; Mice ; Rats ; Rats, Sprague-Dawley ; Recombinant Fusion Proteins ; administration & dosage ; pharmacokinetics