BackgroundLimbal stem cell deficiency usually leads to blindness, and traditional therapy is limited. Recent research demonstrated that bone mesenchymal stem cells ( BMSCs ) and human amniotic epithelial cells(AECs) could differentiate into many kinds of cells including corneal epithelial cells, but the outcome and effect of these cells on corneal stem cell deficiency are still unclear. ObjectiveThis study aimed to observe and compare the effects of rabbit BMSCs and human AECs transplantation for rabbit limbal stem cell deficiency. Methods Eighteen clean New Zealand rabbits were randomly divided into the amniotic stroma(AS) group, rabbit BMSCs group and human AECs group with 6 rabbits for each group. Limbal stem cell deficiency models were established by putting a piece of filter paper that had been soaked in a NaOH solution at the corneal center. Rabbit BMSCs were isolated and purified by density gradient centrifugation combined with the attachment culture method, and human AECs were collected by a sequential trypsin digestion technique,and the third generation rabbit BMSCs and the first generation human AECs were identified with RT-PCR. After that,cells were inoculated onto the denuded AS and grafted to the corneal surface of the experimental animals. Twenty-eight days after cell transplantation, the therapeutic effects were evaluated based on the corneal neovascularization and opacity scores. Corneal histopathological examination and immunohistochemistry were performed to evaluate and compare the effectiveness among AS,rabbit BMSCs and human AECs on corneal stem cell deficiency. The procedure complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission. ResultsThe third generation of rabbit BMSCs grew well after 12 hours, and the first generation of human AECs formed a membrane-like monolayer after 48 hours of incubation on AS. Immunohistochemistry staining showed that, 28 days after transplantation, the surface cells of the cornea were positive for cytokeratin 3 in both the rabbit BMSCs group and human AECs group.Compared with the AS group,the corneal neovascularization and opacity grades were significantly decreased in the rabbit BMSCs group( Z=-2. 983, P =0. 003 ; Z =-2. 844, P =0. 004 ) and human AECs group ( Z =-2. 817, P =0. 005 ; Z =-2.041, P =0. 041 ). Histopathological analysis exhibited that stratified corneal epithelial-like cells formed on the corneal stroms 28 days after grafting and no signs of goblet cells and neovascularization were found. Less inflammatory cells and regular collagen fiber could be seen in the rabbit BMSCs group and human AECs group. In addition,clinical observation also revealed that the corneas were much clearer in the rabbit BMSCs group than the human A ECs group( Z =-2. 091 , P=0. 037 ), but the corneal neovascularization score was similar between them (Z = -0. 267,P=-0. 789). ConclusionsRabbit BMSCs and human AECs can differentiate into corneal epithelial-like cells onthedamagedcornealsurfaceandfurtherdemonstrateremarkableinhibitoryeffectsoncornealneovascularization and inflammatory cells. The more dominant and prominent effect is the role played by rabbit BMSCsin the improvement of corneal transparency.

The existent problems of medical insurance specialty core course system currently are tallied up on the foundation of definition of core course system. The strategy and measure for adjustment are put forward,providing a basis for this professional course reform.

Objective To investigate the association of ret gene rearrangement mutation with the pathogenesis of papillary thyroid carcinoma (PTC). Methods Twenty-seven cases of PTC were analysed for expression of ret gene rearrangement (ret/PTCs) by multiplex-PCR at first, and then ret/PTC1-3 were identified by identification-PCR (ID-PCR). Finally, the specific ret/PTC was affirmed by automated direct sequencing. Ten specimens of malignant thyroid tissues of other histological types, 33 benign thyroid lesions and 30 normal thyroid specimens beside tumor (as the control) were also included. Results (1) Fifteen samples showed positive ret/PTCs, 11 of which harboured ret/PTC1,3 were positive for ret/PTC3, and 1 for ret/PTC2. All the rearrangements were clearly identified by automated direct sequencing of ID-PCR products. (2) All the 15 ret/PTC-positive tissue samples were histologically confirmed to be PTC. The prevalence of the tumor-specific ret rearrangements in 27 patients with PTC is 55.6% (15/27). (3) No significant difference was found regarding the gender and age of the patients, tumor size and metastasis of neck lymph node. 33.3% (2/6) of the PTC with invasion of extrathyroidal soft tissue were ret/PTC-positive, as compared with 66.7%(4/6) for ret/PTC-negative group (P

China ; epidemiology ; Drug Resistance, Bacterial ; Electrophoresis, Gel, Pulsed-Field ; Humans ; Listeria monocytogenes ; physiology ; Listeriosis ; epidemiology ; Serotyping

The purpose of this study was to evaluate the impact of callus induction and culture conditions on secondary metabolic diversity of the callus cell lines of traditional Chinese medicinal plant Glycyrrhiza sp. (Glycyrrhiza) by combined chemical analysis and HPLC fingerprint. These callus induction conditions included two Glycyrrhiza species, two types of explants, light and dark conditions, and two combinations of hormones. The evaluation was firstly based on the contents of total flavonoids in the callus by chemical analysis and one way ANOVA. The content of total flavonoids in callus was significantly (P < 0.05) influenced by Glycyrrhiza species, light condition, and the combination of hormones. The callus was further evaluated using diversity factor based on the comparison of HPLC fingerprints of these callus cell lines. Diversity factor varies significantly for calli induced under different conditions, with the highest being at 0.45 under light condition and combination of hormones. These results provide important knowledge for the selection of suitable callus cell lines for the production of pharmacologically important secondary metabolites or bioactive fractions by in vitro culture of Glycyrrhiza sp.
Cell Culture Techniques ; methods ; Cell Line ; Darkness ; Flavonoids ; biosynthesis ; Glycyrrhiza ; cytology ; drug effects ; metabolism ; radiation effects ; Plant Growth Regulators ; pharmacology ; Plant Roots ; metabolism

Objective To investigate the therapeutic action of Taurine in patients with cerebral infarction(CI)induced by hyperhomocysteinemia(Hhcy).Methods The 100 CI patients induced by Hhcy were randomly divided to two therapy groups.One group was administered Taurine,the other group was administered Folacin combined with Vitamine B12 for 8 weeks.Before and after treating,the serum levels of homocysteic acid(Hcy),Folacin and Vitamine B12 were detected,NIHSS and BI were evaluated.Results The serum levels of Hcy were significantly decreased after administering in both two groups(all P

Amygdala ; drug effects ; metabolism ; Animals ; Conditioning (Psychology) ; Morphine ; adverse effects ; Rats ; Receptors, Opioid, kappa ; metabolism

The experiment explored the influence of glutamic acid (Glu) and the NMDA-receptor antagonist dizocilpine maleate (MK-801) on the pain-evoked responses of pain-excitation neurons (PEN) in the nucleus accumbens (NAc) of rats. The trains of electric impulses applied to the sciatic nerve were used as noxious stimulation. The discharges of PEN in NAc were recorded by glass microelectrode. We observed the influence of intracerebroventricular (icv) injection of Glu and microinjection of MK-801 into the NAc on the noxious stimulation-evoked activities of PEN in NAc. The results showed that the noxious stimulation potentiated the electric activities of PEN in NAc. Intracerebroventricular injection of Glu (10 nmol/10 microl) increased the frequency of the discharge of PEN evoked by the noxious stimulation in NAc, the Glu-induced response was blocked by the injection of MK-801 (1.0 nmol/0.5 microl) into NAc. MK-801 partly inhibited the response of PEN upon the noxious stimulation. It is therefore suggested that the facilitatory effect of Glu on PEN response in NAc to the noxious stimulation is mediated by NMDA receptors, and that Glu and NMDA receptors are involved in the modulation of nociceptive information transmission in the NAc.
Animals ; Dizocilpine Maleate ; pharmacology ; Electric Stimulation ; methods ; Electrophysiological Phenomena ; Female ; Glutamic Acid ; physiology ; Male ; Neurons ; physiology ; Nociceptors ; physiology ; Nucleus Accumbens ; physiology ; Pain ; physiopathology ; Rats ; Rats, Wistar ; Receptors, N-Methyl-D-Aspartate ; antagonists & inhibitors ; physiology

OBJECTIVETo explore the protection of high intensity microwave radiation on hypothalamo-pituitary-adrenal axis (HPAA) activity and hippocampal CA1 structure in rats and the protectiveeffect of Qindan Granule (QG) on radiation injured rats.

METHODSTotally 48 Wistar rats were randomlydivided into 8 groups, i.e., the normal control group, post-radiation day 1, 7, and 10 groups, 7 and 10days prevention groups, day 7 and 10 treatment groups, 6 in each group. Rats in prevention groups wererespectively administered with QG liquid (1 mL/100 g, 4. 75 g crude drugs) for 7 days and 10 days bygastrogavage and then microwave radiation. Then preventive effect for radiation injury was statisticallycalculated with the normal control group and the post-radiation day 1 group. Rats in treatment groupswere firstly irradiated, and then administered with QG liquid (1 mL/100 g, 4.75 g crude drugs). Finally preventive effect for radiation injury was statistically calculated with the normal control group, post-radiation day 7 and 10 groups. Contents of corticotrophin releasing hormone (CRH), beta endorphin (beta-EP), adrenocorticotropic hormone (ACTH), and heat shock protein 70 (HSP70) were detected. Morphological changes and structure of hippocampal CA1 region were observed under light microscope.

RESULTSCompared with the normal control group, contents of CRH and beta-EP significantly decreased in each radiation group. Serum contents of ACTH and beta-EP significantly increased in post-radiation day 1 and 7 groups (P < 0.05). Compared with radiation groups, beta-EP content in serum and pituitary significantly increased, and serum ACTH content significantly decreased in prevention groups (P < 0.05). Pituitary contents of CRH and beta-EP significantly increased in prevention groups. Serum contents of ACTH, beta-EP, and HSP70 were significantly lower in day 7 treatment group than post-radiation day 7 group (P < 0.05). Morphological results showed that pyramidal neurons in the hippocampal CA1 region arranged in disorder, with swollen cells, shrunken and condensed nucleus, dark dyeing cytoplasm, unclear structure. Vessels in partial regions were dilated with static blood; tissues were swollen and sparse. In prevention and treatment groups pathological damage of hippocampal CA1 region was obviously attenuated; neurons were arranged more regularly; swollen, pycnotic, or deleted neuron number were decreased; vascular dilatation and congestion was lessened.

CONCLUSIONQG could affect HPAA function and activity of high intensity microwave radiated rats, showing certain preventive and therapeutic effects of microwave radiated rats by adjusting synthesis and release of partial bioactive peptides and hormones in HPAA, improving pathological injury in hippocampal CA1 region.

Adrenocorticotropic Hormone ; blood ; Animals ; CA1 Region, Hippocampal ; drug effects ; pathology ; radiation effects ; Corticotropin-Releasing Hormone ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; HSP70 Heat-Shock Proteins ; blood ; Hypothalamo-Hypophyseal System ; drug effects ; radiation effects ; Microwaves ; adverse effects ; Pituitary-Adrenal System ; drug effects ; radiation effects ; Random Allocation ; Rats ; Rats, Wistar ; beta-Endorphin ; blood ; metabolism

Aim To investigate the effect of hypercholesterolemia on L-type Ca2+(ICa-L) current and intracellular calcium concentration ([Ca2+]i) in single ventricular myocytes of hypercholesterolemic rats.Methods 12 male wistar rats were randomly divided into two dietary groups:a group fed a normal diet(n=6)and a group fed high-cholesterol diet(n=6) for 4 weeks,respectively. The level of serum lipid was examined.Zymolytic method was used to isolate single ventricular myocytes from hypercholesterolemic and normal rats,which were loaded with Ca2+-sensitive fluorescent indicator Fluo-3/AM.[Ca2+]i represented by fluorescent intensity(FI)was measured by laser scanning confocal microscope.Whole cell patch clamp technique was used to record ICa-L.Results There was no significant influence exhibited on TG level.However, the serum total cholesterol(TC)level of hypercholesterolemic rats was much higher than that of model control group; at the test potential of 0 mV, ICa-L decreased from(-8.56?1.29)pA/pF(Control)to(-5.24?0.90) pA/pF(HC)(n=6 in each group,P