Objective To investigate the incidence and the correlative factors of diabetic retinopathy (DR) in patients with diabetes mellitus(DM)who lives in Beixinjing blocks, Shanghai. Methods Residents with DM were enrolled according to resident health archives. The data of disease history, visual acuity, eye disease and introcular pressure were collected by inquiry and examination. Photography of ocular fundus was used to confirm the diagnosis of DR. Results A total of 535 residents excepted the examination with the participating rate of 90.68%, in whom 146 (27.29%) were identified as with DR. The incidence of single and proliferative DR was 22.29% and 4.30%, respectively. Duration of DM was the independent risk factor of DR, while long duration of DM, accompanied with peripheral neuropathy and body mass index was the in-order independent factor of proliferative DR. Conclusions The incidence of DR is high in residents with DM. Monitoring DR progress in DM residents with risk factors is recommended.

In order to understand the alcohol's toxicity to the quantitative alternations of synapses in mouse visual cortex, the expression of synaptophysin after prenatal alcohol exposure was investigated. In present study, the experimental mice at P0, P7, P14 and P30 were grouped, as control, 2 g x kg(-1) alcohol treatment and 4 g x kg(-1) alcohol treatment. The pre-synaptic elements which were used to represent synapses were marked with synaptophysin (a synaptic vesicle associated protein) by immunocytochemistry technique. The synaptophysin positive boutons in layer VI of visual cortex were imaged under laser confocal microscope. With stereological methods, the number cal density of synapse in visual cortex was calculated in different groups at various ages. Moreover, Western blotting was carried out to detect the expression of synaptophysin in visual cortex. The results showed that prenatal alcohol exposure could cause synaptic loss with long-term effect and in a dose dependent manner. For instance, there were significant difference among the different treatment groups of P0, P14 and P30 as well (P < 0.05). Western blotting supported the results of immunofluorescent labeling. In conclusion, prenatal alcohol exposure can induce the synaptic loss dose dependently and with long-term effect. Our findings implicate that the synaptic loss with long-term effect in CNS probably contributes to the lifelong mental retardation and memorial lowliness associated with childhood FAS.

Objective To quantize the synergistic force in movement of upper limbs between shoulders and elbows. Methods The trans-verse forces of elbows and shoulders during movement were recorded in a healthy adult with an upper-limb-force-measuring plate form which comprised of 2 three-dimensional force sensors, respectively. Then he performed shoulder abduction/adduction and elbow extension/flexion at 100%, 75%, 50%and 25%of the maximum contraction force, respectively. The ratio of the active action force and the joint action force (named assessment index) was used to assess the synergistic degree of the forearm and the upper arm. Results In the shoulder abduc-tion motion, the assessment index decreased as the strength of active action decreased, meant interference of joint action increased. Howev-er, it was almost stable in the shoulder adduction, increased in the elbow extensionas, and was irregular in the elbow flexion, as the active ac-tion strength decreased, respectively. Conclusion It may be more difficult to control upper arm than the forearm.

Objective To investigate the role of cerebellar dentate-interpositus (D-I) nuclei during acquisition of classical eyeblink conditioning in guinea pigs. Methods A 500 ms tone conditioned stimulus (CS) was paired with a 100 ms corneal oxygen-puff unconditioned stimulus (US) in a delay paradigm. Guinea pigs were trained daily to acquire the classically conditioned eyeblink responses. Micorinjections of Muscimol into bilateral cerebellar D-I nuclei were performed during the 4-6 day of training session. A high-resolution potentiometer was used to detect the eyeblink responses. Results The conditioned response (CR) rates of Muscimol-injected group on session 4-6 were significantly lower than their rates on session 3 (P1=0.005,P2=0.004,P3=0.010); Their rates on session 7 and 8 were as the same level as those without microinjection on session 4 and 5 (P1=0.061,P2=0.669). However, there was no significant difference on their amplitude and peak latency of unconditioned responses (URs) during the entire course of training (P1=0.926,P2=0.939). Conclusion The cerebellar D-I nucleus is actively involved in the neural circuitry of motor learning, which is essential for both acquisition and performance of the classically conditioned eyeblink responses.

Objective To observe the adhesion and growth of LLC-PK1 cells and ECV304 cells on titania nanotube arrays, and provide evidence for construction of miniaturation bioartificial kidney. Methods Four different diameters nanotube materials were prepared by anodic oxidation, each material was processed by unannealed and with UV irradiation, annealed and without UV irradiation, annealed and with UV irradiation, respectively, which had 12 groups totally,then two kinds of cells were separately grown on the 12 materials. The adhesion and growth of the two kinds of cells were studied under a fluorescence microscope. MTT assay was used to test the activity of two kinds of cells on different diameters and the proliferation of two kinds of cells on 70 nm diameters. Results The adhesion and proliferation of two kinds of cells on TiO2 nanotube arrays were basically consistent, both on anatase TiO2 nanotubes with 70 nm diameter but without UV irradiation showed the optimal adhesion and activity. The activities of LLC-PK1 cells and ECV304 cells were both increased with time extended, while the absorbance of ECV304 cells was higher on pure Ti film than on titania nanotube. Conclusion TiO2 nanotube is beneficial to LLC-PK1 cells, but is unfavorable for ECV304 cells when they grow alone.

The hippocampus plays a critical role during the consolidation of trace eyeblink conditioned responses (CRs). However, the role of its related structure such as dentate gyms (DG) remains unclear. The present study was aimed at monitoring the activity of single granule cell in the DG during the consolidation of trace eyeblink CRs, and elucidating the possible role of DG during this hippocampus-dependent task. Guinea pigs (n=8) were trained on a trace eyeblink conditioning paradigm using a 200-ms tone conditioned stimulus (CS), a 200-ms corneal airpuff unconditioned stimulus (US) and a 600-ms trace interval. Controls consisted of pseudo- conditioned guinea pigs (n=8). Extracellular single unit recordings in vivo were performed in the DG of learner animals during the consolidation of trace eyeblink CRs. The results revealed that all the trace-conditioned animals acquired the trace eyeblink CRs over 14 training days, however, none of the pseudo-conditioned animals did. Furthermore, 23 of 40 single granule cells in the DG of learner animals exhibited heterogeneous activity patterns during the consolidation of trace eyeblink CRs such as increases in activities to the tone CS, trace interval or airpuff US. The results suggested that the DG might participate in the neural circuit important for the consolidation of trace eyeblink CRs, and that the granule cells might encode different information during the consolidation of trace eyeblink CRs.

Objective To investigate the expression of pSTAT5 in 7 pancreatic carcinoma cell lines,and the change of expression of pSTAT5 in pancreatic carcinoma cells SW1990 after growth hormone (GH) treatment, and explore its molecular mechanism. Methods Human pancreatic carcinoma cell lines (SW1990, Cap-1, Colo, Mia, AsPc, P3, PANC1) were cultured in vitro, and Western blotting was used to detect the expression of pSTAT5 in these cell lines. SW1990 in exponential growth phase was collected and nude Balb/c mice were inoculated with SW1990 cells. When tumors became palpable after inoculation, mice (normal saline group). 1 h, 2 h and 24 h after the last dose of GH treatment, the mice were sacrificed.Western blotting was used to detect the expression of pSTAT5 in SW1990 and inoculation tumor cells after GH injection. Results Positive expression of pSTAT5 was observed in all human pancreatic carcinoma cell lines (SW1990, Cap-1, Colo, Mia, Aspc, P3, PANC1). 5 minutes after GH (50 ng/ml) stimulation, the expression of pSTAT5 in SW1990 was 0.57 ±0.05, which was significantly increased; and it reached 0.64 ±0.04 at 10 minutes, then decreased to 0.39 ±0.03 at 15 minutes, however, it remained higher than that in the control group at 1 h (0.33 ± 0.02 vs 0.25 ± 0.06), and its expression at 2 h was 0.26 ± 0.03 and returned to the normal level. The expression of pSTAT5 in xenograft was not significantly changed. Conclusions GH could rapidly up-regulate the expression of pSTAT5 in SW1990 but the effect lasted for a relatively short period. GH had no significant effect on the expression of pSTAT5 in xenograft.

In order to investigate the role of the PTEN expression in carcinogenesis and development of endometrial carcinoma and clarify whether and how PTEN and PI3K/Akt pathway relate to endometrial carcinoma, the expression of PTEN and phospho-Akt was detected by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) methods and Western-blot from 24 cases of endometrial carcinoma, 10 cases of endometrial atypical hyperplasia, 10 cases of endometrial hyperplasia, and 10 cases of normal endometrium. SP immunohistochemical methods were used to measure levels of PTEN protein expression in following 5 study groups: 31 cases of endometrium in proliferative phase, 30 cases of endometrium in secretory phase, 71 cases of endometrial hyperplasia, 25 cases of atypical hyperplasia and 73 cases of endometrial carcinoma. Immunostaining score of PTEN was 3.39+/-0.15 in proliferative phase, 1.90+/-0.21 in secretory phase, 3.34+/-0.29 in endometrial hyperplasia, 0.62+/-0.11 in atypical hyperplasia, and 0.74+/-0.19 in endometrial carcinoma, respectively. PTEN mRNA relative value in normal endometrium, endometrial hyperplasia, endometrial atypical hyperplasia, and endometrial carcinoma was 2.45+/-0.51, 2.32+/-0.32, 0.46+/-0.11, and 0.35+/-0.13 respectively. The expression levels of PTEN mRNA and protein in patients with endometrial carcinoma and atypical hyperplasia were significantly lower than in those of proliferative phase and with endometrial hyperplasia. The level of PTEN expression in patients with endometrial carcinoma was significantly related to tissue type (P<0.005), differentiation (P<0.05) and clinical stage (P<0.05), but not to depth of myometrium invasion (P>0.05). Western blot analysis revealed that Phospho-Akt level in PTEN negative cases was significantly higher, and there was a negative correlation between PTEN and phospho-Akt (r=-0.8973, P<0.0001). It was suggested that loss of PTEN expression was an early event in endometrial tumorigenesis. The phosphorylation of Akt induced by the loss of PTEN took part in the tumorigenesis and development of endometrial carcinoma.

BACKGROUND:Studies have shown that paeoniflorin functions as replenishing blood and treatment of autoimmune diseases, and bone marrow mesenchymal stem cels also play an important role in the body’s blood and immune function. However, paeoniflorin effects on bone marrow mesenchymal stem cel proliferation and cytokine secretion and expression are rarely reported. OBJECTIVE:To investigate the effect of paeoniflorin on proliferation of bone marrow mesenchymal stem cels and the expression of interleukin-6. METHODS:Human bone marrow mesenchymal stem cels were separated and culturedin vitro by density gradient centrifugation combined with attachment method. The biological characteristics of bone marrow mesenchymal stem cels were identified by flow cytometry and osteogenic/adipogenic induction. The proliferation of bone marrow mesenchymal stem cels under different concentrations of paeoniflorin was detected by MTT method. The mRNA expression and secretion of interleukin-6 in the supernatant of bone marrow mesenchymal stem cels were detected by RT-PCR and ELISA, respectively. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cels were isolated successfuly and had osteogenic and adipogenic differentiation potential. Compared with the controlgroup, 2 μmol/L and 10 μmol/L paeoniflorin significantly promoted the proliferation of bone marrow mesenchymal stem cels. 10 μmol/L paeoniflorin could significantly decrease the proportion of bone marrow mesenchymal stem cels in G0/G1 phase and increase this proportion in S phase. Compared with the control group, the experimental group could significantly increase the secretion and mRNA expression of interleukin-6 (P < 0.01). It is concluded that paeoniflorin at certain concentrations can obviously promote the proliferation of bone marrow mesenchymal stem cels, and increase the expression and secretion of interleukin-6.

Histone deacetylase was overexpressed in a variety of cancers and was closely correlated with oncogenic factors. The histone deacetylase inhibitor, trichostatin A (TSA) was shown to induce apoptosis in many cancer cells. However, the mechanism of TSA on induction of cancer cells apoptosis is poorly understood. This study was designed to characterize the global gene expression profiles before and after treatment of human leukemia cell line Molt-4 with TSA. Flow cytometry, MTT and DNA ladder were used to observe the effect of TSA on the apoptosis of MOLT-4 cells and normal human peripheral blood mononuclear cells (PBMC). Microarray, reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were used to detect the difference of gene and protein expressions of Molt-4 cells after incubation of the cells with TSA. The results showed that TSA could induce Molt-4 apoptosis in dose- and time-dependent manners but spared PBMCs. Microarray analysis showed that after incubation with TSA for 9 h, 310 genes were upregulated and 313 genes were deregulated. These genes regulate the growth, differentiation and survival of cells. Among these genes, STAT5A was down-regulated by 80.4% and MYC was down-regulated by 77.3%. It was concluded that TSA has definite growth-inhibiting and apoptosis-inducing effects on Molt-4 cells in time- and dose-dependent manners, with weak cytotoxic effects on PBMCs at the same time. The mechanism of TSA selectively inducing apoptosis and inhibiting growth may be ascribed to the changes of pro-proliferation genes and anti-apoptosis genes.