Objective To observe the protective effect of progesterone(PROG) on rat retinal ganglion cells(RGCs) in chronic ocular hypertention models.Methods The chronic ocular hypertention rat model was made by cauterizating three episcleral veins.Rats were divided into control group,high level PROG group,middle level PROG group,low level PROG group according to different concentrations of PROG injected intraperitoneally.The left eye was model eye and the right eye was control eye.Three months later,the animals were executed and the eyeballs were enucleated.The RGCs were detected by HE staining and Thy-1.1 immunohistological staining.The apoptosis of RGCs was detected by TdT-dUDP terminal nick end-labeling(TUNEL) technique.Results The number of RGCs of model eyes in high level PROG injection group was more than those in control group,low level and middle level groups(P

Aim To construct human phage single-chain antibody library associated with esophageal cancer and to screen the specific scFv against Eca109 cells from the liberary. Methods Metastatic periesophageal lymph nodes of esophageal cancer patients were used as the B cells source, the total RNA of these B cells was extracted and prepared as the template of RT-PCR. First, we screened graticulely two pairs of primers of the heavy and light regions separately, then the V_H and V_L fragments were first amplified from the cDNA by the polymerase chain reaction (PCR). Second, the V_H-linker and V_L-linker were amplified from the V_H and V_L fragments. Last, the V_H-linker and V_L-linker were assembled into scFv gene fragments by SOE-PCR,and then Sfi I and Not I restriction site were inlet in it. ScFv gene was cloned into the pCANTAB-5E phagemid. Phagemids were introduced into E.coli TG1 by electrotransformation, followed by rescue of antibody-expressing phage using M13K07 helper-phage superinfection. Recombinant scFv phage library was constracted and PCR was used to identify the insert ratio of scFv antibodies library. Results of SfiI/Not I double digestion reaction positive insert clone were identified by 1.5% agarose gel electrophoresis. The phage library was panned with NHEEC and Eca109 cancer cells in suspension for four rounds. Strongly positive recombinant phage clones were used to infect E.coli HB2151. Expression of soluble scFv was induced by IPTG. Soluble scFv from periplasm were purified by affinity chromatography and identified by SDS-PAGE and Western blot. Cell ELISA , immunohistochemical staining and immunocytochemical staining were used to identify the activity of the soluble scFv. Results The result of agarose gel electrophoresis showed that total RNA of these B cells had two bands of 28 S and 18 S. The size of V_H fragment is about 450 bp,V_L fragment is about 350 bp and scFv is about 850 bp. The competence is 108 cfu??g-1 pUC18 DNA. Randomly digestive reac-tion showed that the positive insert ratio was 91.7% (22/24). After four rounds of panning, the fourth phage yield is 141 times as much as that of the first one. SDS-PAGE and Western blot showed that the MW of the soluble scFv was about 30 ku and the brand of 30 ku was stained. Immunohistochemical staining showed strong stainning of the tissue of esophageal cancer, but not the liver and gastric cancer tissue. Immunocytochemical staining showed significant staining of the esophageal cancer line Eca109. The result of cell ELISA assay revealed that soluble scFv had highly specific and could combined with Eca109 cells, but not with BGC-823 and NHEEC. Conclusion A human scFv phage display library associated with esophageal cancer has been constructed successfully and the specific scFv antibody against Eca109 has been identified from the liberary.

OBJECTIVE: To evaluate the inhibitive effect of matrine modification X on the growth of human nasopharyngeal carcinoma CNE2 cell xenografts in nude mice. METHOD: Tumor model was established by subcutaneous inoculation of nasopharyngeal carcinoma cell CNE2 into nude mice, which was used to evaluate the antitumor effect of matrine modification X in vivo. The expression levels of Bax, Bcl-2, Caspase3 were detected by real-time PCR and western blot. RESULT: The growth of xenografts in nude mice was significantly suppressed after application of matrine modification X in a dose-dependent manner. The inhibition rates were 32.55% and 44.89% when treated at medium and high dose respectively. Real-time fluorescence quantitative-PCR and Western Blot results showed that the expression of Bax and Caspase3 increased, while the expression of Bcl-2 decreased in a dose-dependent manner. The change of high dose group was obvious, and the difference was statistically significant (P < 0.05). CONCLUSION Matrine modification X could significantly inhibit the growth of human nasopharyngeal carcinoma CNE2 cell xenografts in nude mice, probably by inducing the apoptosis of nasopharyngeal carcinoma cells, and the possible mechanism is related to regulating the expression level of Bax/Bcl-2 and Casepase3.
Alkaloids ; pharmacology ; Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; Carcinoma ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Heterografts ; Humans ; Mice ; Mice, Nude ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Quinolizines ; pharmacology ; bcl-2-Associated X Protein ; metabolism

Objective To explore the characteristics of patients with stagnation of liver-qi syndrome using brain functional imaging techniques.Methods10 patients with stagnation of liver-qi syndrome and 20 normal persons whom paired by sex and age were scanned with positron emission tomography(PET).Data analysis was carried out using SPM in MATLAB software.ResultsCompared with normal,some cerebral areas of the patients with stagnation of liver-qi syndrome showed glucose metabolism letdown(P<0.001),including right parietal lobe(postcentral gyrus,BA1/2),temporal lobe(middle temporal gyrus,BA37),frontal lobe(middle frontal gyrus,BA9/10)etc.,while some cerebral areas showed glucose metabolism heighten(P<0.001)including right frontal lobe(superior frontal gyrus,inferior fontal gyrus,BA22/28/44),right cingulated gyrus,double midbrain and cerebella,etc.ConclusionThe function of some cerebral areas can be changed undergoing stagnation of liver-qi syndrome.

Objective To investigate the clinical significance of flow cytometry (FCM) assay in following up of the minimal residual disease (MRD) used for predicting relapse and guiding chemotherapy. Methods The clinical data of 43 acute leukemia patients diagnosed by MIC were collected in our hospital from 2005 July to 2008 June.Bone marrow aspirates were collected from 43 patients with newly diagnosed acute leukemia after induction therapy and during constimulation therapy. The cells with leukemia associated with immunophenotype were investigated using FCM, as immunologic target of MRD. Results MRD were detected earlier in predicting the relapse than those of the traditional bone marrow cells morphology assay by an average of 4-6 months. The results of the MRD following up: MRD was negative at CR in 26 cases, 6 cases relapse, 20 cases of them were kept negative during following up. MRD was positive in 17 cases at CR, 9 cases of them were relapse. 4 cases after intensified chemotherapy the MRD became negative and kept egative for more than one year. The MRD of the 43 cases at CR were divided into 3 groups, MRD less than 1×10-4 group (A group) MRD between 5×10-3 and 1×10-4 group (B group) and MRD above 5×10-3 group(C group). By chi square test. There was no statistical significance between A group and B group, but there was tatistical significance between B group and C group (P=0.02). Conclusion The application of FCM in detecting MRD has important clinical significance in predicting relapse and guiding chemotherapy.

Objective To investigate the changes of non-linear dynamics characteristics of electroencephalogram(EEG) in patients with the stagnation of Liver-Qi syndrome.Methods 15 Liver-Qi stagnation syndrome patients and 15 normal persons were paired by sex and age.The data of EEG under three states(eyes closed,eyes opened,and mental arithmetic with eyes opened) were analyzed by the parameter of point-wise correlation dimension(PD2).Results PD2 of the stagnation of Liver-Qi syndrome group increased significantly compared with that of the healthy control group and always maintain high level.The difference of PD2 between two groups was gradually reduced along with the increase of stimulated condition.Conclusion Non-linear dynamic analysis is more appropriate for the study of mental functions changes of stagnation of Liver-Qi syndrome and can be used to investigate the brain mechanism of the stagnation of Liver-Qi syndrome.