Objective To study the expression of telomerase and its possible clinical significance in patients with condyloma acuminatum,bowenoid papulosis,Bowen's disease and vulvar squamous cell car- cinoma.Methods The human telomerase reverse transcriptase (hTERT) was detected by immunohisto- chemistry technique in all patients.Histopathological analysis was done in patients with bowenoid papulosis or Bowen's disease.Results There was a significant difference in the expression of hTERT among normal tissue,condyloma acuminatum and squamous carcinoma,and the expression rate increased successively from normal control to squamous carcinoma.In bowenoid papulosis,the expression of bTERT was significantly higher as compared to that in condyloma acuminatum,significantly lower as compared to that in squamous carcinoma,and was as the same as that in Bowen's disease.Histopathologically,the degree of cell atypia was significantly higher in Bowen's disease than that in bowenoid papulosis.Conclusions The expression of telomerase increases successively from normal skin,benign tumors to malignant carcinomas,which sug- gests that the expression of telomerase may play an important role in both cell proliferation and immortaliza- tion.

Objective To explore the expression levels of microRNA(miR)-34a in hippocampus of temporal lobe epilepsy rats and the effect of miR-34a on its target signaling pathway Notch1.Methods Rats were divided randomly into experiment group (n=40) and control group (n=40) by adopting random number table method.The status epilepticus model and the temporal lobe epilepsy model were induced by using lithium-pilocarpine for experiment group.The control group rats received an injection of an equal amount of 9 g/L saline as instead of pilocarpine.Racine grading was performed at 24 hours,day 3,day 7,day 15,and 1 month after modeling to evaluate the behavior.Real-time fluorescent quantitative polymerase chain reaction was used to test the mRNA expressions of miR-34a and Notch1.Western blot was performed to explore the protein expression of Notch1.Results The expression levels of miR-34a at post-status epilepticus in 24 hours,day 3,day 7,day 15 were 2.55±0.29,2.11±0.17,1.68±0.49 and 1.84±0.42,respectively,which showed statistically significant difference compared with the control group (1.00±0.00) (t=-1.55,-1.11,-0.68,-0.84,all P<0.05).The expression levels of Notch1 mRNA at post-status epilepticus in 24 hours,day 3,day 7,day 15,1 month were 1.44±0.31,1.27±0.13,1.52±0.28,0.91±0.33,and 0.80±0.09 respectively.There were significant differences at 24 hours,day 7 in Notch1 mRNA expression (t=-0.44,-0.52,all P<0.05) compared with the control group(1.00±0.00).The expression level of Notch1 mRNA on day 15 was significantly lower than 24 hours and day 7 (t=-0.54,-0.62,all P<0.05),and the expression in 1 month was significantly lower than in 24 hours,or day 3 and day 7 (t=-0.64,-0.46,-0.72,all P<0.05).The expression levels of Notch1 protein at post-status epilepticus 24 hours,day 3,day 7,day 15,1 month were 0.78±0.09,0.57±0.13,0.55±0.16,0.42±0.13,and 0.33±0.09,respectively.There was significantly up-regulated at 24 hours of Notch1 protein expression compared with control group (0.51±0.15)(t=-0.20,P<0.05);and the expression level at day 15 were significantly lower than 24 hours (t=-0.26,P<0.05),while the expression in 1 month was significantly lower than in 24 hours and on day 3 (t=-0.36,-0.24,all P<0.05).Conclusion miR-34a is significantly up-regulated in the post-status epilepticus rat hippocampus,and it may contribute to temporal lobe epilepsy by activating Notch1 signaling pathway.

Objective To observe effects of ginsenoside Rb1 on phosphorylation of microtubule-associated protein 2 (pMAP-2) in hippocampus and amygdala of depressive model rats. Methods Thirty male Wistar rats were randomly divided into control group, model group and treatment group. The depression rat model was produced by giving chronic unpredicted mild stress (CUMS). Treatment group was given daily intragastric administration of ginsenoside RB 1 (1 g/mL crude drug, 1 mL/100 g body weight) for 22 days during modeling. Western blot assay was used to detect expressions of MAP-2 and pMAP-2 protein, and real-time PCR was used to detect expressions of pMAP-2 mRNA respectively. Results The expressions of pMAP-2 protein and mRNA in hippocampus and amygdala were significantly lower in model group than those of control group (P < 0.05). The expressions of pMAP-2 protein and mRNA were significantly higher in treatment group than those of model group (P<0.05). Conclusion Ginsenoside Rb1 can play anti-depression role by inhibiting the phosphorylation of MAP-2 in rats.

Objective To establish a method of PKH26 labeled bone marrow-derived endothelial progenitor cells (EPCs) into rats with liver fibrosis and observe cell immigration and differentiation in the liver after transplantation.Methods Bone marrow-derived EPCs were isolated and cultured from rats with liver fibrosis,and then labeled with PKH26 in vitro.Under the scanning confocal microscopy and flow cytometry,PKH26 fluorescent labeling rate and cell survival rate were measured.EPCs of PKH26 fluorescent labeling were transplanted into rats with liver fibrosis via the tail vein,and the migration situation was observed in the liver.Endothelial cell markers CD31 and von willebrand factor (vWF) were detected by using immunofluorescence.Results The PKH26-labeled EPCs appeared red fluorescence and the labeling rate was 96.65 ％.As compared with unlabeled cells,the labeled cells grew well,and had no significant changes in the growth curve.After transplantation into the liver of rats,the PKH26 labeled cells were mainly distributed in blood vessel endothelium along fibers and hepatic sinusoids in hepatic lobule.Endothelial cell-specific antigens such as CD31 and vWF could be detected along the vascular walls.Conclusion PKH26 could be used to label and track EPCs in the liver of rats with liver fibrosis in vitro.The PKH26-labeled cells may migrate to the surrounding of the hepatic vessels and differentiate into mature endothelial cells.

Objective To compare the advantages and values of several special staining methods of chondrocytes . Methods Twelve 7-day old healthy C57BL/6J mice were killed to obtain the cartilage tissue of the knee joint in order to isolate the chondrycytes .Type II collagen was used to assess the chondrocytes .Then the chondrocyte climbing slices were prepared.The materials were fixed, and HE staining, Safranin O-fast green staining, SA-β-gal staining and immunohistochemical staining of Type II collagen were performed and compared .Results HE staining showed clear morphology of the chondrocytes .The cell nuclei were stained blue and the cytoplasm was pink .Safranin O-fast green staining showed that the nuclei were pink and the cytoplasm green .SA-β-gal staining showed that the aging cells were green while the young cells were colorless .Immunohistochemical staining of type II collagen showed the distribution of type II collagen and they were stained brown while the cell nuclei were blue .Conclusions HE staining and safranin O-fast green staining can provide more information than the other staining techniques .SA-β-gal staining is useful in the analysis of aging chondrocytes .Immunohistochemical of type II collagen can be used to study type II collagen .

OBJECTIVETo investigate the effects of intrathecal injection of ginsenoside Rg1 at different doses on the changes of the behavior and the expressions of excitatory amino-acid transporter 1 (EAAT1), i. e., glutamate-aspartate transporter (GLAST) in the spinal dorsal horn of the arthritis rats with chronic morphine tolerance, and further to explore its mechanisms for morphine tolerance.

METHODSAfter successful intrathecal injection, an adjuvant arthritis model was established in 36 healthy male SD rats. They were randomly divided into 6 groups, 6 in each group. They were intrathecally injected with 10 microL normal saline (Group NS), 10 microg morphine (Group M), 10 microg morphine + 50 microg ginsenoside Rg1 (Group MG50), 10 microg morphine +100 microg ginsenoside Rg1 (Group MG100), 10 microg morphine + 200 microg ginsenoside Rg1 (Group MG200), and 100 microg ginsenoside Rg1 (Group G100), respectively. The normal saline and morphine were intrathecally injected twice daily, while ginsenoside Rg1 at different doses was intrathecally injected once daily, for 7 successive days. Fifty percent mechanical paw withdrawal threshold (PWT) was dynamically detected to evaluate their behaviors. The rats were sacrificed on day 7 after medication. The L3-L5 segment of the spinal cord was isolated for determining the expression of GLAST in the spinal dorsal horn using immunofluorescence staining.

RESULTSThe PWT of Group M was significantly higher than that of Group NS on the 1st and 3rd day after medication (P < 0.05). But it was gradually shortened along with the increasing days of medication. There was no statistical difference between Group M and Group NS on the 7th day (P > 0.05), indicating the formation of morphine tolerance. The PWT of Group MG100 also showed a decreasing tendency, but obviously slower than that of Group M (P < 0.05). The PWT of Group G100 was higher than that of Group NS (P < 0.05). Compared with Group NS, the expression of GLAST in the spinal dorsal horn of rats in Group M was down-regulated (P < 0.01). Compared with Group M, the expression of GLAST in the spinal dorsal horn of rats in Group MG100 and Group G100 was up-regulated (P < 0.05).

CONCLUSIONSSingle application of ginsenoside Rg1 showed mild antinociceptive effect in adjuvant-induced arthritis rats. Intrathecal injection of 100 microg ginsenoside Rg1 could attenuate the formation of morphine tolerance. Its mechanisms might be correlated with up-regulating of the expression of GLAST.

Amino Acid Transport System X-AG ; metabolism ; Animals ; Arthritis, Experimental ; metabolism ; Drug Tolerance ; Ginsenosides ; administration & dosage ; pharmacology ; Injections, Spinal ; Male ; Morphine ; pharmacology ; Pain Measurement ; Rats ; Rats, Sprague-Dawley

The discovery of drug targets plays a crucial role in drug research. Accurate information of small molecule drug-protein interaction can be provided by label-free target discovery technology without any structural modification at the small molecule. So, the label-free drug target discovery technology had become the powerful tool to discover the targets of drugs. Due to the “multi-component and multi-target” characteristics of traditional Chinese medicines (TCMs), the research on its targets and mechanism had been restricted. Based on potential of the label-free target discovery technology in the research of TCMs, this paper summarized the label-free target discovery technology and its application in TCMs research. It will provide a reference for the discovery of targets of TCMs and a new view for promoting the modernization of TCMs.

Objective To investigate the factors associated with hyporesponsiveness to erythropoietin (EPO) in patients on maintenance peritoneal dialysis (PD) . Methods Data of 114 PD patients in our PD center were collected . Patients were divided into three groups according to weekly EPO dose: hyperresponsive, hyporesponsive and normal responsive . Various factors were compared among three groups by linear correlation and ordinal regression analysis to predict EPO resistance . Results As compared to hyperresponsive and normal responsive groups,significantly lower serum hemoglobin [(78 .11±13 .42) vs (106 .28±23 .83), (96 .31±12 .33) g/L],albumin [(33 .98±4 .78) vs (39 .72±4 .26), (35 .76±4 .88) g/L], and significantly higher serum CRP [(26 .08±21 .66) vs (5 .46±1 .75), (11 .82±5 .63) mg/L], ferritin [(371 .08±89 .38) vs (289 .39±76 .84),(323 .07±62 .46) μg/L] were found in hyporesponsive group(all P ＜ 0 .01) . Erythropoietin resistance index (ERI) was correlated with CRP and albumin . Regression analysis showed that serum albumin,CRP and ferritin were strong predictors of EPO hyporespnsiveness . Conclusions Serum albumin,CRP and ferritin are closely related to hyporesponsiveness . Malnutrition and inflammatory state contribute to EPO hyporesponsiveness .

ET-CT demonstrated that VX2 tumor tissues could uptake 18F-FDG more than normal tissue, which made the basis for further study of VX2 tumor model.

Objective To observe the effects of fosinopril(FOS),a new generation angiotensin-converting enzyme inhibitor(ACEI),on protein and mRNA expression of transforming growth factor-?_1(TGF-?_1) of rat glomerular mesangial cell(GMC) induced by lipopolysaccharide(LPS);to demonstrate the preventive mechanism against glomerular sclerosis by applying FOS.Methods The cultured GMC in classic way were divided into 3 groups:control group;LPS group;LPS+FOS group.TGF-?_1 concentration in GMC supernatant fluid was detected by ELISA;TGF-?_1 mRNA expression was determined by semiquantitative real-time RT-PCR.Results LPS group was obviously higher than control groups in TGF-?_1 secretion and mRNA expression,while LPS+FOS group decreased distinctively in TGF-?_1 secretion and mRNA expression compared with LPS group.Conclusions FOS has obviously inhibited on TGF-?_1 expression of rat GMC both at protein level and mRNA level,which reveals that it may be an important mechanism by FOS on restraining the development of glomerulosclerosis.