Objective To observe the opticin expression in the eyes of non-obese diabetes (NOD) mice and non-diabetic NOD mice . Methods Twenty NOD mice were divided into diabetic group (experimental group) and non-diabetic group (control group). All the mice were killed by cervical dislocation method. The eyes were harvested, and the vitreous, retina and sclera were separately collected. Western blot and real-time reverse transcription-polymerase chain reaction(RT-PCR)were respectively used to determine opticin protein and OPTC-mRNA levels. Results The opticin protein level in the vitreous and retina was lower in the experimental group(t = 4.42,4.58; P = 0. 002,0. 002), but is same in the sclera between the 2 groups (t = 0. 27, P = 0. 794). OPTC-mRNA level was vitreous> retina> selera. OPTC-mRNA levels of vitreous and retina in diabetic group were significantly Iower(t = 3.30,2. 48; P= 0. 01, 0.04); there was no statistical significant on OPTC mRNA of sclera between two groups(t = 0. 27, P = 0. 80). Conclusion Expression of opticin was suppressed in retina and vitreous of diabetic mice.

Objective To construct small hairpin RNA (shRNA) expression plasmid targeting rat opticin gene. Methods Four pairs of opticin oligonucleotides were synthesized and inserted into the plasmid vector, resulting into four plasmids: shRNA-1, shRNA-2, shRNA-3 and shRNA-4. Then the four constructed shRNA expression vectors and empty vector were transfected into rat ciliary non-pigment epithelium (NPE) cells by lipofectmaine 2000. Non-transfected NPE cells were set as control group. The expression of opticin mRNA and protein were measured by Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot respectively. Results The opticin mRNA expression of the shRNA-1,shRNA-2, shRNA-3, shRNA-4 group were decreased compared with the control group (F = 10. 239, P = 0. 000);the inhibitory rate were 85.7% ,62. 87% ,54.87% and 48.77% respectively. The opticin protein expression of the shRNA-1,shRNA-2, shRNA-3, shRNA-4 group were also decreased compared with the control group (F=17.870, P= 0.000);the inhibitory rate were 78.7%, 34.6%, 31.1% and 16.8% respectively.Conclusions The shRNA-1 expression plasmid has most potent inhibitory effect on opticin expression in rat ciliary NPE cells.

ObjectiveDahl salt-sensitive (Dahl/SS) rats are hereditary salt-sensitive hypertensive rats.Its pathogenesis is similar to that of human primary hypertension,CPB established in Dahl/SS rats provides an animal model for the study of CPB in patients with primary hypertension.MethodsMale 14-16 weeks old Dahl/SS rats weighing 360-390 g were fed with high salt (8％ NaCl) diet for 4 weeks before the experiment.Ten Dahl/SS rats were randomly divided into 2 groups ( n =5 each) according to the CPB time:groups Ⅱ and Ⅲ underwent CPB for 120 and 75 min respectively.Another 7 male 14-16 weeks old ordinary SD rats weighing 410-490 g undergoing CPB for 120 min were used as control group (group Ⅰ ).Anesthesia was induced with isoflurane inhalation.Orotraeheal intubation was performed.The animals were mechanically ventilated.Right jugular vein and tail artery were cannulated for venous drainage and arterial inflow from CPB circuit.Blood was oxygenated with a customized mini-oxygenator.Blood gases were analyzed and blood glucose concentration was determined.MAP was recorded before (baseline) and at 30 and 60 min of CPB and 30 and 90 min after CPB.The rate of changes in MAP and blood glucose concentration and survival rate at 7 d after termination of CPB were recorded.ResultsThere was no significant difference in blood gases among the 3 groups.The rates of change in MAP and blood glucose concentration were significantly higher during and after CPB in Dahl/SS rats than in control SD rats in a duration of CPB dependent manner.The survival rate at 7 d after CPB was 7/7 (in group Ⅰ ),1/5 (in group Ⅱ ) and 4/5 (in group Ⅲ ) respectively.ConclusionA model of 75 min CPB is established successfully in Dahl/SS rats.

Objective To analyze the long-term results of fractionated stereotactie radiotherapy(FSRT)for the local residual lesion after the first course of radiotherapy for nasopharyngeal carcinoma.Methods From July 1997 to July 2002,46 patients were treated with FSRT.According to the 1992 Fuzhou staging system,the number of patients was 1,6,30 and 9 with stage Ⅰ,Ⅱ,Ⅲ and Ⅳ disease,respectirely;3,11,27 and 5 with T1,T2,T3 and T4 tumor,respectively;14,16,12 and 4 with N0,N1,N2and N3 disease.Radiotherapy was delivered to tumors with the total of dose 68-70 Gy in 7-8w.Chemotherapy(2 cycles of PVF or POF)was given to the patients with stage Ⅲ and Ⅳ a disease.FSRT was given to the residual disease with the total dose of 18-24 Gy in 3 fractions with an interval of 3-7 days.The reference dose line was 70%-90%.Resuits CR and PR rates in this group were 61%and 39%,respectively.The overall survival rates of each year from 1- to 5-year were 100%,87%,83%,78%and 76%.The 1-,3- and 5-year disease-free survival rates were 100%,93%and 89%;The distant metastasis-free survival rates were 100%,85%and 79%;The local-regional control survival rates were 100%,94%and 91%.Seventeen patients who died during the follow-up period were 1 for local cervical lymph node recurrence,2 for fatal nasopharyngeal hemorrhage,4 for local nasopharynx recurrence,and 10 for distance metastases. Conclusions Fraetionated stereotactic radiotherapy is safe and effective for the patients with residual lesion of nasopharyngeal carcinoma at the primary site after radiotherapy.The optimized fractionation and total dose requires the further investigation.

Objective To study the expression status of estrogen receptor-alpha36(ER-α36)in breast cancer tissue and its rela-tionships with the occurrence,development and clinical prognosis of breast cancer.Methods 653 cases of breast cancer tissues were selected in this study.The real-time reverse transcription polymerase chain reaction(RT-PCR)and immunohistochemistry were used to detect the expression of ER-α36,estrogen receptor-alpha66 (ER-α66),progesterone receptor(PR)and human epidermal growth factor receptor-2(Her-2).The relationships between the expression of ER-α36,ER-α66,PR and Her-2 and the pathological charac-ter were analyzed.Results The expression rate of ER-α36 in all cases was 40%.The expression rate of ER-α36 in Her-2 positive tissues(63%)was significantly higher than that in the Her-2 negative group(44%,P <0.05).The expression rate of ER-α36 in ER-α66/PR/Her-2 negative tissues(66%)was significantly higher than that in the non-three-negative group(35%,P <0.05).The differences of ER-α36 expression rate between ER-α66 positive samples and negative samples or between PR positive and negative samples showed no statistical significance(P >0.05).The expression rate of ER-α36 in stage Ⅲ+Ⅳ breast cancer tissues(54%) was significantly higher than that in stage Ⅰ+Ⅱ breast cancer tissues(28%,P <0.05).The expression rate of ER-α36 in breast cancer tissues with lymph node metastasis (55%)was significantly higher than that in breast cancer tissues without lymph node me-tastasis (23%,P <0.05).Conclusion The results indicate that ER-α36 may play a very important role in the occurrence,develop-ment and lymph node metastasis of breast cancer,and be associated with the expression of Her-2,breast cancer staging and lymph node metastasis.ER-α36 is expected to become a new tumor marker and clinical diagnosis and treatment target.

Objective To investigate the meridians and acupoints selection in the treatment of post-stroke spastic paralysis with Tuina (Chinese massage). Methods The literatures about Tuina therapy for post-stroke spastic paralysis were retrieved. The frequency of meridians and acupoints used was counted and analyzed. Results 99 papers were collected, which involved in 226 acupoints and 1602 times of application. The selected acupoints were distributed in all the fourteen meridians and 64.5% (1033/1602, 606 on upper limb, 427 on lower limb) of them were on the limbs. The acupoints of the Yang meridians was 75.0% (1200/1602) and the specific acupoints was 71.7% (1148/ 1602). Conclusion The acupoint selected was basically focused on the local areas in Tuina for post-stroke spastic paralysis, assisted with the involved meridians and distal acupoints. The acupoints of the Yang meridians were the first option, mainly on the extremities. The specific acupoints were the major components of the prescription, especially the Five-shu acupoints and the He acupoints.

Objective To identify differentially expressed N-linked glycoproteins between hepatocellular carcinoma ( HCC) and adjacent non-tumorous liver tissues .Methods N-linked glycoproteome was extracted by multi-lectin affinity chromatography comprising concanavalin A (ConA), lentil lectin (LCH), and snowdrop lectin (GNA) and subsequently subjected to two-dimensional electrophoresis ( 2DE ) and mass spectrometry ( MS ) for identification of differential glycoproteins between 10 pairs of HCC and adjacent non-cancer tissue .Western blotting was used to verify different expression of human liver carboxylesterase 1 (hCE1), haptoglobin (HP)and cathepsin D (CD).Invasion potential in vitro was examined after si-RNA mediated CD gene scilencing .Results LC-ESI-MS/MS identified a total of 28 differentially expressed glycoproteins (14 up-regulation and 14 down-regulated).Western blotting detected consistent down-regulation of hCE1 and HP, and up-regulation of pro-cathepsin D (pCD) in HCC.Up-regulation of ConA-binding CD (ConA-CD), however , was verified in HCC only after ConA-CD enrichment by ConA chromatography .Down-regulation of CD expression mediated by CD-siRNA markedly inhibited the in vitro invasive potential of SNU449 and SNU473.Conclusion Dysregulation of HP , hCE1 expression and alteration of glycans linked to CD may play crucial roles in pathogenesis of HCC.

Objective The nano-zirconia scaffolds we previously prepared had a good 3-dimensional ( 3D ) connectivity but did not achieve the ideal sintering rate and compressive strength .The objective of this study was to explore the enhancing effect of polyvinyl butyral ( PVB) as a dispersant on the compressive strength of 3D nano-zirconia porous scaffolds for bone tissue engineering . Methods We prepared the slurry containing different concentrations of PVB and ana-lyzed the improving effect of PVB on the mechanical properties of the scaffolds by sediment experiment , compressive strength test and scan-ning electron microscopy . Results The sediment experiment showed
no significant stratification in the slurry with 0.2wt%PVB, white suspension in the upper layer and white precipitate in the lower layer , with a significantly higher compressive strength of the scaffold ([0.324 ±0.030] MPa) than that of the scaffold prepared by adding other concentrations of PVB to the slurry (P <0.01).And the compressive strength of the scaffold constructed by adding no dispersant ([0.109 ±0.021] MPa) was remarkably lower than that of the scaffold constructed by adding PVB to the slurry (P<0.05).Scanning electron microscopy demonstrated that the scaffold prepared by adding 0.2wt%PVB to the slurry had a complete porous structure with the fewest and most sparsely distributed surface cracks as compared with other PVB concentration groups . Conclusion PVB can signifi-cantly improve the stability of zirconia slurry , enhance the compressive strength of the nano-zirconia porous scaffold , and make the scaf-fold more applicable to bone tissue engineering .

Objective:To examine the expression of odontoblast related proteins in dental pulp stem cells(DPSCs)induced by BMP-7.Methods:DPSCs were cultured in the common culture medium or medium supplemented with 1 00 ng/ml BMP-7.Electron microscope,CCK8 and immunohistochemical staining were carried out to estimate the cell morphology and differentiation.Results:In-duced by BMP-7,the morphology of DPSCs was not changed,the proliferation of DPSCs was slower than that of the cells without BMP-7 treatment.DPSCs were negative for the expression of DSPP,DMP-7 and ALP.However,DPSCs were found strongly positive for DSPP,DMP-7 and ALP after the induction of BMP-7.Conclusion:BMP-7 induction may promote the differentiation of DPSCs.

In the present study, the total RNA was extracted from three instar larvae of Musca domestica, the cDNA sequence encoding the ORF of cecropins was amplified by RT-PCR, and the target fragment was further sequenced after being cloned into T vector pUCm-T. Then, the cDNA sequence of the mature cecropins was amplified by PCR with recombinant plasmid pUCm-T/cecropin as template, the N-terminal rare codon GGA of E. coli was changed to the favorable codon GGC,and a Asn codon AAC was added in front of the stop coden TAA in the C- terminus. This mutant gene designated as mCecropin was then ligated with the fusion expression vector pGEX-4T-1. After restriction analysis and DNA sequencing, the positive recombinant plasmid pGEX-4T-1/mCecropin was transformed to different strains of E. coli cells and the fusion protein was expressed after IPTG induction. The fusion protein was assayed by SDS-PAGE and the E. coli BL21(DE3) cell was chosen as the host cell for the expression of the fusion protein. The expressed fusion protein GST-mCecropin was purified by GSTrap affinity coloum and the GST marker was then cleaved by thrombin. In this way, the fusion protein mCecropin with antibacterial activity was obtained after purification with HiTrap benzamidine column.