Background Recurrence of pterygium is a common complication after the surgical excision of pterygium,and this procedure is related to cell proliferation,inflammation and neovascularization.Researches determined that rosiglitazone can suppress inflammation and neovaseularization and inhibit proliferation,hut few studies concerning the effect of rosiglitazone on pterygium were performed. Objective The aim of this study was to investigate the effect of peroxisome proliferator-activated receptor γ agonist on the proliferation and apoptosis of human pterygium fibroblasts(HPFs)in culture and search for a new drug to prevent and cure the recurrence after pterygium surgery. Methods Human pterygium samples were obtained during surgery and HPFs were cultured and purified using an explant method and 0.25%trypsin digestion,respectively.The identity of cultured HPFs was confirmed by immunohistochemistry using anti-vimentin and keratin antibodies.Rosiglitazone with the concentrations of 0(control),5,10,25,50,75,100,150,200,400μmol/L was then added in the culture medium for 12,24 or 72 hours.1%DMSO was used as blank control.The MTT method was used to assay the biologic effects of rosiglitazone on HPFs.Cell cycle distribution and apoptosis of HPFs after rosiglitazone treatment were studied by flow cytometic analysis.The expression of proliferating cell nuclear antigen(PCNA)mRNA in HPFs was detected by real-time PCR. Result Cultured HPFs radially migrated outward from the pterygium block,and then grew in long fusiform shape,showing positive response for vimentin and negative for keratin.The HPFs became round and thin with loose distribution after the addition of rosiglitazone.Following 25-125 μmol/L rosiglitazone administration for 12,48 or 72 hours,the A490 value of HPFs significantly declined with the increase of dosage(F=158.312,P=0.006)and lapse of time(F=1.924,P=0.135).Following the treatment of 25,75 or 125 μmol/L rosiglitazone for 24 hours,the number of HPFs in G0/G1 phase was markedly elevated;while the cell numbers in S phase decreased significantly in comparison with the control group(P＜0.05).The apoptotic rate of HPFs in the 25,75 and 125 μmol/L rosiglitazone groups significantly increased with the increase of rosiglitazone concentration(P＜0.05).Real-time PCR revealed that after 24 hours of rosiglitazone treatment,the expression of PCNA mRNA in HPFs was suppressed in a dose-dependent manner(F=3244.329,P＜0.05). Conclusion Rosiglitazone inhibits HPFs proliferation,arrests their cell cycle progression in G0/G1 phase,induces apoptosis of HPFs and depresses the synthesis of PCNA in a dose-and time-dependent manner.

Objective To analyze the mechanism of fentanyl combines midazolam,by comparison with some of the commonly used anesthetic method to determine in the ethical advantages of anesthesia.Methods 45 rats were randomly divided into three groups,Group A:1％ anesthetized by intraperitoneal injection of sodium pentobarbital 40 mg/kg; Group B:5 ％ ketamine 120 mg/kg intraperitoneal injection; Group C:midazolam 5 mg/kg + fentanyl 0.05 mg/kg Last intravenous anesthesia.Observe the effects of anesthesia.Results The rats which are used of fentanyl conbines midazolam anesthesia,the onset of anesthesia time:0.02 ± 0.03 min,maintenance of anesthesia time:89.73 ± 22.59 min,the the narcotic operation time:1.51 ± 0.30 min,the success of anesthesia cases:15 cases.Conclusions Fentanyl combines midazolam compared with the currently used method of anesthesia,the advantages of the more prominent,it has significant improvement and protection about the security,utilization and recovery ethics in experimental rats.

The cupping spot is considered as one kind of skin change due to cupping treatment. With literature regarding cupping spot, the influencing factors and value of cupping spot in clinical diagnosis and treatment were analyzed, which could make a further exploration on the action mechanism of cupping treatment. The literature showed that the formation of cupping spot was related with cupping temperature, pressure, cup-retaining time, cupping area, individual difference and health condition, etc; cupping spot had the ability to assist diagnosis, prevent disease, cure disease and evaluate clinical efficacy. Previous studies on cupping spot have already made some progress, and played a positive significance on finding cupping rule and studying its mechanism. However, the research for this area is still in the primary stage, which needed deeper study to reveal scientific connotations of cupping spot.
Acupuncture Therapy ; instrumentation ; Humans ; Skin ; anatomy & histology ; blood supply

Objective:To observe the effect of herbal cake-partitioned moxibustion on the expressions of mitogen-activated protein kinase (MEK1/2) and extracellular regulatory protein kinase (ERK1/2) in gastric tissues of rats with spleen deficiency syndrome, and to explore the possible mechanisms of herbal cake-partitioned moxibustion in treating spleen deficiency syndrome. Methods:Sixty Sprague-Dawley (SD) rats were randomly divided into a blank control group (group A), a model group (group B), a ranitidine group (group C), and a herbal cake-partitioned moxibustion group (group D) by random digit, 15 rats in each group. Rat models of spleen deficiency syndrome were made by intragastric administration of 4℃ 200% concentrated Da Huang (Radix et Rhizoma Rhei). After successful modeling, the rats in group C were treated with 25 mg/(kg·bw) ranitidine by intragastric adminstration and rats in group D were treated with herbal cake-partitioned moxibustion at Zusanli (ST 36) and Zhongwan (CV 12), for 8 d. Excepted for rats in group A, all the other rats were treated with indomethacin at 5 mg/(kg·bw) at 8:00 a.m. on the second day after finishing all the intervention and sacrificed 7 h later to isolate the stomach. Histopathological changes of the gastric tissues were observed under light microscope after hematoxylin-eosin (HE) staining. The protein expressions of MEK1/2 and ERK1/2 in the gastric tissues were detected by immunohistochemistry. Results:After intervention, the gastric mucosal injury in group B was significantly severer than that in group A, with large breakage and ablating; the damage of gastric mucosa was decreased in group C compared with group B; the gastric mucosal surface remained relatively complete, and the status of breakage and ablating was significantly improved. After intervention, compared with group A, the protein expressions of MEK1/2 and ERK1/2 in gastric tissues of the other groups were significantly higher (P<0.01). Compared with group B, the protein expressions of MEK1/2 and ERK1/2 in group C and D were significantly higher (allP<0.01). Compared with group C, the protein expressions of MEK1/2 and ERK1/2 in group D were significantly higher (P<0.01). Conclusion: Herbal cake-partitioned moxibustion promotes the repair of gastric mucosa in rats with spleen deficiency syndrome, via improving protein expressions of MEK1/2 and ERK1/2 in gastric tissues, as well as activating MEK/ERK signaling pathway.

A variety of natural and artificial cryoprotectant extenders have been explored to enhance sperm recovery following cryopreservation-thawing process.The current investigation is aimed at evaluating the effect of acetyl-L-carnitine on human spermatozoa and reactive species oxygen (ROS) level after freezing-thawing process.The spermatozoa were collected from 35 male patients diagnosed as having asthenospermia.The cryopreservation of human spermatozoa treated with acetyl-L-camitine at different concentrations (group B:2.5 mmol/L,group C:7.5 mmol/L,group D:15 mmol/L) was compared with control (group A:no acetyl-L-carnitine given).For the frozen-thawed spermatozoa,the viability,motility and DNA integrity were measured by comet assay,acrosome integrity by FITC-PNA staining and ROS level was determined in each group.The results showed that there were no significant differences in motility and viability between group A and group B,while the motility and viability of spermatozoa in group C and group D were significantly increased as compared with those in group A.As compared with group A,the values for DNA integrity parameters including comet rate (CR),tail DNA percentage (TD),tail length (TL) and Oliver tail moment (OTM) were significantly reduced in group C and group D.Group C and group D also displayed a higher proportion of intact acrosome than group A.No significant difference in ROS level was found between group A and group B,while with the increase in acetyl-L-camitine concentration,the ROS level in groups C and D was significantly reduced as compared with that in group A.In conclusion,acetyl-L-camitine at a concentration of 7.5 mmol/L is an effective antioxidant against cryo-damage on post-thawed human spermatozoa.

OBJECTIVETo study the effect and mechanism of mexican tea herb and pilular adina herb (abbreviated to MP) on concrescence of gastric mucosa in experimental gastric ulcer rats by observing the changes of epidermal growth factor (EGF), nitrogen monoxidum (NO) and expression of epidermal growth factor receptor (EGFR).

METHODSThe rat ulcer model was established by 100% glacial acetic injection into the subserosa. The ulcer index (UI) was measured by sliding caliper. The levels of NO and EGF in tissue and serum were measured by the nitrate reductase method and enzyme-linked immunosorbent assay, respectively. The expression of EGFR in the mucosa around the ulcer was detected by the immunohistochemical assay and microimage analysis system.

RESULTS(1) Compared with the model group, UI of MP groups (10, 15 and 20 mg.kg(-1).d(-1)) and ranitidine group was lower (P<0.05 or P<0.01), the levels of NO and EGF in the tissue and serum were higher (P<0.05), the thickness of regenerated mucous membrane increased, and the width loss of lamina muscularis mucosa decreased (all P<0.05). (2) The expression of EGFR is weakly positive in gastric mucosa cells in the normal group, mainly in the cytoplasm and cytomembrane. In the model group, the expression of EGFR was mainly in epithelial cells in cervical part and basilar part of gastric gland around the ulcer margin, and the number of cells with EGFR weakly positive expression was more than that in the normal group. Compared with that in the normal and model groups, the number of cells with EGFR positive in MP groups and ranitidine group increased (all P<0.05), with weakly positive expression.

CONCLUSIONMP can protect gastric mucosa, cure gastric ulcer, restrain the secretion of gastric acid, and boost multiplication, differentiation, migration and repair of the endothelial cell by promoting the secretion of NO and EGF, and increasing the expression of EGFR of gastric mucosa epithelial cells.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Epidermal Growth Factor ; analysis ; Female ; Gastric Mucosa ; chemistry ; drug effects ; pathology ; Immunohistochemistry ; Male ; Nitric Oxide ; analysis ; Rats ; Rats, Sprague-Dawley ; Receptor, Epidermal Growth Factor ; analysis ; Regeneration ; Stomach Ulcer ; drug therapy ; pathology ; Tea

Objective To investigate the metabolic alterations in the brain of neonates with HIE and correlate those alterations with clinical grading and prognosis of HIE.Methods Fourty-six eases of full-term neonates diagnosed as HIE clinically were performed MRI and 1~H-MRS,9 healthy neonates without the evidence of asphyxia were studied as controls,1~H-MRS techniques included single voxel proton MRS and two dimensional muhi-voxel chemical shift spectroscopy imaging,point resolved spectroscopy sequence was used for 1~H-MRS.Metabolic changes in the spectroscopy were analyzed in neonates with HIE,and study the relationgship between MRS findings and prognosis.Results(1)The typical 1~H-MRS manifestations of full- term neonates suffering from HIE were as follows:the peaks of Lac were elevated,GLx-? were elevated and NAA were decreased.(2)GLx-?/Cr ratio in control,mild,moderate and severe HIE group was 0.16, 0.21,0.64,and 1.31,respectively.Lac/Cr ratio in control,mild,moderate and severe HIE group was 0.12,0.14,0.19,and 0.26,respectively.There was a significant difference in the ratio of GLx-? and Lac/Cr between HIE group and control group(t=5.01,P

OBJECTIVETo clone the full-length Rcet3 gene, a novel gene related to family 2 cystatins, from mouse testis or other tissues.

METHODSRcet3 gene was cloned using digital differential display (DDD) and RT-PCR was performed for cloning the full-length Rcet3 gene from adult mouse testis cDNA library with sequence analysis.

RESULTSRcet3 cDNA was 610 bp in length, consisting of 4 exons to encode a protein with 140 amino acid residues. The encoded protein contained a potential signal peptide and a cystatin domain, but lacked critical consensus site important for cysteine protease inhibition. These characteristics could be seen in the Cres subgroup related to the family 2 cystatins. Rcet3 was specifically expressed in adult mouse testis, epididymis and the cerebrum, but at higher levels in the testis than in the epididymis and cerebrum.

CONCLUSIONRcet3 may be a new member of Cres subgroup of family 2 cystatins.

Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; Cystatins ; genetics ; DNA, Complementary ; chemistry ; genetics ; Gene Expression Profiling ; Male ; Mice ; Mice, Inbred C57BL ; Molecular Sequence Data ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Testis ; metabolism

OBJECTIVETo study the effect of laminarin sulphate (LAMS) on the expressions of PTEN and P27kip1 in androgen-independent prostate cancer PC-3 cells in vitro, and investigate the mechanism of its anti-tumor action.

METHODSThe inhibitory effects of different concentrations of LAMS (0, 50, 100, 200 microg/ml) on androgen-independent prostate cancer PC-3 cells were detected by WST-8 assay. The morphology of PC-3 cells was observed under the fluorescence microscope, and the cell cycle and apoptosis were analyzed by flow cytometry. The mRNA and protein levels of PTEN and P27kip1 were measured by RT-PCR and Western blot.

RESULTSLAMS inhibited the proliferation of androgen-independent prostate cancer PC-3 cells in a dose- and time-dependent manner, and the cell cycle analysis showed that PC-3 cells were arrested in the S phase after treated with different concentrations of LAMS. The rate of apoptosis was increased and many typical apoptotic morphological features were observed under the fluorescence microscope. The PTEN and P27kip1 expressions at mRNA and protein levels were increased in a dose-dependent manner.

CONCLUSIONLAMS can inhibit the proliferation, arrest the cell cycle in the S phase and induce apoptosis of prostate cancer PC-3 cells. The significantly increased expressions of PTEN and P27kip1 may be one of the mechanisms for LAMS inhibiting prostate cancer PC-3 cells.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p27 ; genetics ; Humans ; Male ; PTEN Phosphohydrolase ; genetics ; Polysaccharides ; pharmacology ; Prostatic Neoplasms ; blood ; genetics ; RNA, Messenger ; genetics

The present study aimed to investigate the protective effect and mechanism of hydrogen sulfide donor NaHS administration against gastric mucosal injury induced by gastric ischemia-reperfusion (GI-R) in rats. GI-R injury was induced by clamping the celiac artery of adult male SD rats for 30 min and followed by reperfusion for 1 h. The rats were randomly divided into sham group, GI-R group, NaHS group, glibenclamide group and pinacidil group. Gastric mucosal damage was analyzed with macroscopic injured area, deep damage was assessed with histopathology scores, and the hydrogen sulfide concentration in plasma was determined by colorimetric method. The results showed that pretreatment of NaHS significantly reduced the injured area and deep damage of the gastric mucosa induced by GI-R. However, NaHS did not significantly alter the levels of hydrogen sulfide in plasma 14 d after NaHS administration. The gastric protective effect of NaHS during reperfusion could be attenuated by glibenclamide, an ATP-sensitive potassium channel (K(ATP)) blocker. However, K(ATP) opener pinacidil inhibited the GI-R-induced injury. These results suggest that exogenous hydrogen sulfide plays a protective role against GI-R injury in rats possibly through modulation of K(ATP) channel opening.
Animals ; Gastric Mucosa ; pathology ; Hydrogen Sulfide ; metabolism ; Ischemic Preconditioning ; methods ; KATP Channels ; metabolism ; physiology ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control ; Stomach ; blood supply ; Sulfides ; pharmacology