0.05). There was no correlation between Bcl-2 and Fas positive cells and acetylcholine receptor antibody serum titre, disease grouping, age and sex in patients with or without hyperplastic thymic and thymoma.Conclusions In MG patients, the expression of Bcl-2 in hyperplastic thymus is upregulated. There is no correlation between the expression of apoptotic genes Bcl-2 and Fas and clinic relative factors.

BACKROUND:Corneal hemangiogenesis occurs in 40%-60%patients after keratoplasty.Blood vessel is one of the high risk factors for corneal immunological rejection.To inhibit corneal hemangiogenesis would prolong the survival time of the grafts and promote the successful rate of the keratoplasty.OBJECTIVE:To explore the inhibitive effects of doxycycline on corneal angiogenesis after keratoplasty.DESIGN,TIME AND SETTING:A randomized controlled animal experiment was performed at the State Key Laboratory of Ophthalmology(No.2006DA105054),Zhongshan Ophthalmic Center,Sun Yat-sen University from March to August 2007.MATERIALS:A total of 48 healthy dean Sprague Dawley rats served as recipients(right eye)and 24 Wistar rats as donors(both eyes).CD31-PEfluorescent antibody was obtained from Sigma,USA.Sandwich enzyme-linked immunosorbent assay(ELISA)kit for vascular endothelial growth factor(VEGF)was brought from RapidBio,USA.METHODS:Corneal allogenic transplantation models were established in rats.Recipients were equally and randomly divided into 2 groups:saline control group and doxycycline group.Twenty minutes prior to surgery,mydriasis was performed using 1%atropine,with a diameter of 2.75 mm of implant and 2.5 mm of implant bed.In the saline control group,conjunctiva of the right eye received saline,three times a day,following surgery.In the doxycycline group,conjunctiva of the right eye received 1%doxycycline,three times a day,till 30 days following surgery.MAIN OUTCOME MEASURES:The following parameters were measured:corneal angiogenesis using immunofluorescence,expression of VEGF protein by using ELISA.RESULTS:Compared with the survival time of saline control group[(9.67±2.73)days],the mean survival time of doxycycline group[(20.67±3.01)days]was significantly prolonged(P＜0.01).The mean percentages of neovascularized corneal area in the saline control group were(4.00±1.00)%,(14.33±4.04)%,(31.33±3.51)%at 3,7 and 14 days after keratoplasty,respectively.The mean percentages of neovascularized corneal area in the doxycycline group were(1.67±1.15)%,(4.67±1.53)%,(18.33±1.53)%at the same time point respectively.Compared with the saline control group,the mean percentages of neovascularized corneal area of the doxycycline group was significantly reduced at 7 and 14 days after keratoplasty(P ＜0.05).The expression of VEGF in the saline control group was(541.00±75.44)pg/mg,(960.00±90.14)pg/mg,(976.00±130.41)pg/mg at 3,7 and 14 days after keratoplasty,respectively,while expression of VEGF in the doxycycline group was(115.33±9.29)pg/mg,(239.00±41.62)pg/mg,(361.00±65.20)pg/mg,respectively.The difference of VEGF expression at all time points between the two groups was significant (P＜0.05).CONCLUSION:Doxycycline has a significant effect in inhibiting corneal angiogenesis and prolonging survival time of implants after keratoplasty.

Objective To investigate the influence of different HBV DNA extraction methods with Micro-Nucleic-Releaser (MNR), polysaccharide deposition method and boiling method in real-time quantitative and provide reference data for clinical lab regarding the PCR kit selection. Methods Three different HBV DNA isolation methods including MNR, polysaccharide deposition method and boiling method were used to study the extraction efficiency and anti-interference ability when HBV DNA virus in lipemia, haemolysis or jaundice serum samples was detected with real-time PCR. The traditional HBV DNA isolation method of Hydroxybenzene-Chloroform was used as control method. Results The quantitative results and amplification efficiency differed among different extraction methods. Complete hemolytic sample brought biggest influence, modest hemolytic sample give minimal influence, whereas the lipemia and jaundice did not bring significant influence to detection. The MNR give the best amplification efficiency and quantitativereproducibility. The extraction efficiency of boiling method was lowest. The fluorescent efficiency of real-timePCR from Hydroxybenzene-Chloroform methods was better than other methods, but it suffered from losingHBV DNA. Conclusions Real-time PCR system of HBV DNA isolation by MNR can provide an accurate,highly sensitive, and rapid method to quantify Hepatitis B virus. It is suitable to use in clinical laboratory.

ObjectiveTo observe the effects ofYushen Zhuyun Prescription on ovarian morphology and sex hormone of rats with decreased ovarian reservation (DOR) induced by tripterygium glycosides; To explore its mechanism of action.Methods Tripterygium glycosides solution was used to establish DOR model rats by gastric gavage. 48 female SD rats were randomly divided into blank group, model group, positive medicine group andYushen Zhuyun Prescription low-, medium- and high-dose groups.Yushen ZhuyunPrescription groups were given the low, medium, high dosagesYushen Zhuyun Prescription liquid; positive medicine group were given estradiol valerate liquid; model group and blank control group were given normal saline with the same amount, by gavage 2 times per day for 14 consecutive days. The general condition of the rats was observed; ovarian was weighed and the viscera index was calculated; ovarian morphology was observed by HE staining; the levels of serum estradiol (E2), follicle stimulating hormone (FSH) and testosterone (T) were detected by radioimmunoassay; the levels of inhibin B (INHB) and anti Mullerian hormone (AMH) were detected by ELISA.Results Compared with the blank group, ovarian tissue atrophied; the number of follicles was reduced; ovarian index and the level of E2, INHB and AMH decreased; the levels of FSH and LH increased in the model group (P<0.01). Compared with the model group, ovarian tissue morphology improved significantly; the number of follicle and corpus luteum increased; follicular atresia was reduced; ovarian index and the levels of E2, INHB, and AMH increased; the levels of FSH and LH decreased in Yushen Zhuyun Prescription high-dose group (P<0.01). INHB and AMH had significant correlation (P<0.01). ConclusionYushen Zhuyun Prescription can regulate hormone levels, promote the growth and secretion of follicle, and inhibit follicular atresia, thereby improve ovarian reserve function.

BACKGROUND:Buccal Multiloop Removable Appliance can interceptively correct mutiple adolescent malocclusions. But the clinical problem of Buccal Multiloop fatigue fracture is not solved yet. How to prolong the fatigue fracture cycle is stil in the research stage. OBJECTIVE: To study the effect of different temperature of heat treatment on the Buccal Multiloop fatigue fracture cycle, thereby to select a relatively optimal method to enhance the fatigue fracture cycle. METHODS: Thirty-five left HL-2 Buccal Mltiloops were divided into seven groups according to different ways of heat treatment. Each group consisted of five samples. They were an untreated group, three pre-bending groups (320, 420, 520℃ heat treatment before bending) and three post-bending groups (320, 420, 520℃ heat treatment after bending). The dental stainless steel wires and Buccal Multiloop were respectively treated by low-temperature annealing. The data were recorded and evaluated after the samples tested by the 3D Simulating Movement of TMJ Testing Machine. The features of fatigue fracture were observed by scanning electron microscope. RESULTS AND CONCLUSION: The mean values of the Buccal Multiloop fatigue fracture cycle from largest to smalest were as folows: 520℃ pre-bending group > 420℃ pre-bending group > 320℃ pre-bending group > untreated group > 520℃ post-bending group > 320℃ post-bending group > 420℃ post-bending group. The fatigue fracture cycle of Buccal Multiloop made of the dental stainless steel wires after 520℃ annealing treatment was longer than others. By the observation of scanning electron microscope, the fracture crack extension area had the tendency to expand, transient interruption was delayed and the tissue structure became more uniform.

Objective To explore the expression and significance of HDGF and ADAM9 in esophageal cancer.Methods HDGF and ADAM9 expression in 113 patients with esophageal cancer (78 males and 35 females) with ages ranging from 28 to 87 years,averaged (61.2 ± 8.6) years,were analyzed by SP immunohistoehemical staining.Results The positive rates of HDGF and ADAM9 in esophageal cancer (78.8％ and 68.1％ respectively) were significantly higher than those in normal tissue(all were 30％).ADAM expressions were not correlated with age,sex,tissue types,lymph nodes metastasis,location and tumor size (P ＞ 0.05) ;the positive expression rate of HDGF and ADAM9 was related to differentiation,PT stages,and 5-year survival rate (P ＜0.05).HDGF was correlated with PT stage.Conclusion The expressions of HDGF and ADAM9 in esophageal cancer tissues were more higher than normal esophageal tissues,this means they could promote the development of tumor cells transformation,multiple and moving,and may be an useful tool for providing information about the targeted therapy and prognosis.

Objective To investigate the effect of cysteine-rich protein 61 (Cyr61) on proliferation and cell cycle in human renal tubular epithelial cells (HK-2).Methods Cyr61 cDNA was cloned into pEGFP-N2,then HK-2 cells were transfected with the recombinant plasmid pEGFP-N2-Cyr61 by Lipofectamine.The cell proliferation was measured by MTT.The expression level of Cyr61,p-FAK and cyclin dependent cyclin-dependent kinase 2 (CDK2) protein were detected by Western blotting.The cell cycle and cell apoptosis were analyzed by flow cytometry.Results The recombinant plasmid pEGFP-N,-Cyr61 could be transfected into HK-2 efficiently.After transfection,the proliferative activity was significantly increased,the proportion of HK-2 cells in G1 phase decreased and in S-phase increased significantly,the level of cell apoptosis decreased markedly (all P ＜ 0.01).The expressions of Cyr61,p-FAK and CDK2 in Cyr61-transfected group were all amplified significantly (all P ＜ 0.01).Conclusions Cyr61 protein over-expressed in HK-2 cells can increase CDK2 expression throngh FAK pathway,resulting in the promotion of HK-2 cells entering into S phase,cell proliferation and the reduction of cell apoptosis.

Objective To investigate the effect and mechanism of cysteine-rich protein 61 (Cyr61) on oxidative stress in human kidney tubular epithelial cell line after anoxia.Methods Human kidney tubular epithelial cell line (HK-2 cells) were divided into 5 groups:control group,Cyr61 group,MAPK inhibitor group (Cyr61 +PD98059),p38 inhibitor group (Cyr61 +SB203580) and PI3K inhibitor group (Cyr61+Wortmannin).Each group was pretreated for 12 h and then injured by anoxia.The cell viability was determined by MTT assay and the apoptosis rate of HK-2 cells was determined by flow-cytometry.The cellular ROS level was measured by spectro-fluorometry.The cellular superoxide dismutase (SOD) and catalase (CAT) were measured by nephelometry test.The expression of Nrf2 in HK-2 cells was detected by Western blotting.Results Anoxia enhanced the expression of ROS and Nrf2,decreased the expression of SOD and CAT significantly,meanwhile decreased HK-2 viability and increased HK-2 apoptosis (all P ＜ 0.05).Cyr61 increased the expression of p-Akt,Nrf2,SOD and CAT in HK-2,and decreased the expression of ROS,at the same time increased HK-2 viability and decreased HK-2 apoptosis (all P ＜ 0.05).Wortmannin inhibited the expression of p-Akt,Nrf2,SOD and CAT,meanwhile decreased HK-2 viability and increased HK-2 apoptosis (P ＜ 0.05).PD98059 and SB203580 had no affect on HK-2 compared to Cyr61 group (P＞0.05).Conclusions Cyr61 promotes the expression of Nrf2 through PI3K pathway in HK-2,which enhances the expression of SOD and CAT,and decreases the expression of ROS.Cyr61 exhibits protective effects on HK-2 cells injured by oxidative stress after anoxia.

AIM: To investigate the effects of lidocain and gentamycin on inflammation reaction and wound infection of explosive wound in limb arthrosis after seawater immersion. METHODS: The explosive wound were made in 48 New Zealand white rabbits. All rabbits were randomly divided into 4 groups: lidocain, gentamycin and glutin sponge bound up and immersed in seawater 1 h group (LGGS group, n=12); bactroban and glutin sponge bound up and immersed in seawater 1 h group (BGS group, n=12); 12 layers common gauze bound up and immersed in seawater 1 h group (GS group, n=12) and 12 layers common gauze bound up 1 h group (G group, n=12). The changes of blood WBC, CRP, IL-1, IL-6, TNF-?, MDA and SOD in each group were tested. The bacterial culture was performed in each group after explosion 1 h, 3 d, 7 d and 1 month. RESULTS: Compared with pre-experiment, the WBC was decreased, the IL-6 and TNF-? values were increased significantly (P0.05), but the other groups did (P

Objective To clarify the clinical significance of visit-to-visit variability in blood pressure (BP) of stage 3-4 chronic kidney disease (CKD) patients with hypertension.Methods One hundred and fifty-two cases of stage 3-4 CKD patients with hypertension were enrolled in the study.Variability in BP was defined as the standard deviation (SD) in BP.For each patient,SD and mean BP from BP measurements were calculated at all the visits.Correlations between the decline in estimated glomerular filtration rate (eGFR) and SD in BP were analyzed by multivariable regression.Results Visit-to-visit variability in BP was significantly associated with renal function decline (P ＜ 0.05),in addition,baseline eGFR,baseline albuminuria and mean SBP during follow-up were significantly associated with renal function decline as well (all P ＜ 0.05).The percentage of CCBs used in low SD of the SBP group was higher than that in high SD of the SBP (76.1％ vs 58.2％,P ＜ 0.05).Conclusion Visit-to-visit variability in BP is significantly associated with renal function decline.Drugs which can decrease the variability of blood pressure should be the first choice in the treatment of hypertension.