Background Establising the culture model of Müller cells for obtaining the highly putified target cells is essential for the study about the physiology and pathology of retinal Müller cells. The exsiting purifing method for culturing Müller cells is dissatisfactory. Objective This study was to establish a method to obtain high purifing Müller cells. Methods The retina from 5 clean newborn SD rats were isolated and digested by 0. 01% trypsin and cultured in DMEM containing 10% fetal bovine serum. The cellular suspension was then prepared,and the target cells were screened using flow cytometry based on the size and the quantity of cells. Cultured and passaged cells were identified by transmission electron microscope and light microscope. Immunocytochemistry was used to detecte the expression of GFAP in cultured cells for the determination of type and purity of the cells. Results The cells showed the similar shape to retinal Müller cells after primarily culture with the large volume, and some small other types of cells could been seen. The growth of cells was quickly 3 weeks later. The fibroblasts were removed using sticking-wall by steps,and neurons were eliminated following passage. Aboundent of cellular organs were seen under the transmission electron microscope. The positive response rate of the cells for CFAP was 100%. Conclution Flow cytometry offer a rapid and feasible approach for purifying Muller cell and it builds the foundation for further study about Müller cells.

AIM: To understand the relationship between pentanucleotide repeat(PNR) polymorphism of the apolipoprotein(a) gene and lipoprotein natures of normal Han population. METHODS: The serum levels of TG, TC, LDL-C,HDL-C, apo AI, apo B and Lp(a) were measured and the polymorphism of the apo(a) PNR was studied by using PCR-SSCP in 153 random normal individuals in Hubei Han population respectively. RESULTS: The relative frequencies of apo(a) PNR allele were significantly different from western population. The apo(a) gene which copy number of PNR is 5 was associated with high Lp(a) levels. No marked differences in the levels of TG, TC,LDL-C, HDL-C, apo AI and apo B were found among the various genotype groups of apo(a) PNR in Hubei Han. CONCLUSION: The data of lipids and PNR polymorphism of the apo(a) gene from healthy Hubei Han were obtained. The apo(a) allele with 5(TTTTA)-repeats may be related to high serum Lp(a) levels in the Hubei Han population.

Aim To establish a model of the vascular endothelial cell (VEC) injury in SDrats.Methods SD rats were randomly divided into the control and the modelgroups. The model rats were injected with adrenaline diluted to 2. 5 times 0. 05 mg?100 g-1 (tid) for 5 d continously. From the 4th d, they were irritated for 5 min in the0℃ cold-water in the middle between adrenaline injections.The control rats weregiven 0. 9% NS as above. At 6th d, blood samples were taken from carotid arteries ofthe rats and the CEC counts, t - PA、PAI activities, 6-keto-PGF1? concentrations andthe platelet aggregation rate(max) were detected respectively. Results In the modelgroup, as compared with those in the control group, t - PA activity and 6-keto-PGF1?concentration decreased significantly(P

Objective To evaluate the clinical effects of thoracic close drainage with thin drainage tube assisted to thick drainage tube after video-assisted thoracic surgery(VATS)lobectomy. Methods We ret-rospectively reviewed 89 patients received VATS lobectomy in Chinese PLA General Hospital from January 2014 to September 2014. The patients with non-small cell lung cancer were divided into two groups:treatment group (50 patients)and control group(39 patients). Treatment group took thin tube assisted to thick tube of thoracic close drainage and control group took general thoracic closed drainage tube. We studied the operation time,the bleeding of operation,the number of lymph node dissection,time of first activity out of bed,the hospitalization time of post-operation,post-operative complications,the days of post-operative drainage,drainage volume,the effect of drainage,the VAS evaluation score of post-operative pain in the two groups. Results Compared with control group,there was no statistical significance in the differences of the time of operation[(2. 58 ± 0. 57)h vs(2. 57 ± 0. 50)h;t = 0. 127,P = 0. 681],bleeding of operation[(108. 00 ± 52. 84)ml vs(114. 10 ± 107. 18)ml;t = 0. 352,P = 0. 334],the number of lymph node dissection[(14. 20 ± 5. 95)vs(11. 21 ± 4. 71);t = 2. 576,P = 0. 068)],the staying time of drainage[(5. 66 ± 2. 53)d vs(5. 82 ± 2. 02)d;t =0. 324,P = 0. 219],the postoperative drainage volume[(1 141. 76 ± 819. 26)ml vs(1 022. 95 ± 464. 84) ml;t = 0. 889,P = 0. 367]and the occurrences of the post-operative complications(8. 00% vs 10. 25% ;χ2 =1. 750,P = 0. 726). There was statistical significance in the differences of the post-operative time of off-bed [(11. 28 ± 8. 78)h vs(13. 97 ± 7. 83)h;t = 4. 027,P = 0. 045],the time from surgery to discharge [(8. 36 ± 2. 63)d vs(9. 56 ± 2. 89)d;t = 2. 952,P = 0. 043]and the drainage effect(costophrenic angle sharp:72. 0% vs 46. 2% ;χ2 = 5. 329,P = 0. 017). In the two groups,there were statistical significance differences in scores of VAS for the 24 to 72 hours resting and coughing of post-operation:24 h[(2. 78 ± 1. 13)vs(3. 74 ± 1. 68);t = 3. 226,P < 0. 001)],48 h[(1. 98 ± 0. 59)vs(3. 33 ± 1. 72);t = 5. 189,P <0. 001)],72 h[(1. 94 ± 0. 55)vs(3. 15 ± 1. 60);t = 5. 010,P < 0. 001)],coughing[(3. 64 ± 1. 23)vs (5. 33 ± 1. 95);t = 5. 005,P < 0. 001)]. Conclusion The thin drainage tube assisted to thick drainage tube for thoracic close drainage make the drainage more effective,release the pain,shorten the hopital stay;moreo-ver,it is simple and safe for operation and easy to popularize with high modified value.

The M and NP genes of H5N1 avian influenza virus (A/chicken/Hubei/489/2004) were amplified by RT-PCR from viral RNA,and cloned into pMD 18-T vector respectively.The expression plasmid containing the M gene (pHM6-m) or the NP gene (pHM6-np) was then constructed by inserting the M or NP gene into the pHM6 eukaryote expression vector; the constructed plasmid was then sequenced.32 BALB/c mice (6-week-old) were divided into four groups at random.Three groups of BALB/c mice were inoculated one time the intramuscular route with either 30 μg of plasmid pHM6-m,30 μg of plasmid pHM6-np or the mixture of plasmid pHM6-m (15 μg ) and pHM6-np(15 μg) respectively.A additional group of mice were injected with 100 μ1 PBS as controls.Two weeks later,all mice were challenged with homologous H5N1 avian influenza virus,and observed in the following 12 days.The survival rates of mice in the pHM6-m group,the pHM6-np group and mixed plasmids group were 62.5% ,25.0% and 50.0%,respectively.Results showed that effective protection could be provided by either pHM6-m or pHM6-np,but pHM6-m provided a better protective effect than pHM6-np.

Objective To investigate the morphology and relationships with the adjacent structures in the pineal region on the thin sections and to provide anatomic data for imaging diagnosis and surgical treatment of diseases in this region. Methods By CT and MRI examination, one normal head specimen was selected for this study. Using the computerized freezing milling technique, the specimen was sliced from anterior to posterior. The in vivo MR images were obtained from ten normal Chinese male adult volunteers using a 3.0 T GE scanner. The base lines of the sectioning and the MR scan were perpendicular to the AC-PC line. Then primary sections were contrasted with the corresponding MR images. Results By the appearance of the pineal peduncle and the disappearance of the pineal gland, the pineal region could be divided into three parts from anterior to posterior, and the shape changed from an inverted triangle to a trapezoid and a triangle gradually. The first interspace was getting wider in the anterior and middle parts of the pineal region, while in the posterior part of the pineal region, it was getting narrower and disappeared finally. From anterior to posterior, the bilateral internal cerebral veins were always in the midline of the pineal region and descended gradually.Conclusion By the computerized freezing milling technique, the anatomic details and adjacent relationships of the pineal region could be exhibited clearly in the thin serial sections, which could help the imaging diagnosis and surgical treatments for minute diseases in this region.

Objective To investigate the affect of body positions on biochemical indexes. Methods By autogenous contrast and cross matched survey, 107 volunteers divided into 3 season patches of winter, spring and summer, blood samples were drawn from the same part in both standing and lying positions。From19 persons, blood samples were collected respectively after standing and sitting for 15 min, lying for 15 min and 30 min and then sitting for another 15 min。 The blood samples were analyzed for 32 biochemical indexes on analyzer。Results 25 biochemical indexes in sitting position were significantly different from those in lying position (P0。05)。Conclusions Changing body position can result in obvious physiological variation of 28 biochemical indexes, particularly of those related to protein. Such result may lead to abnormality in some marginal values. It suggests body position should not be neglected in analyzing laboratory data.

Objective: To explore the relationship between mental symptoms and functional dyspepsia (FD), and the effect of Xiaoyu decoction (XYD) plus psychotherapy on FD. Methods: SCL-90 scale and FD symptom scale were used to estimate the condition of 56 healthy subjects and 56 patients of FD before and after 4 weeks treatment with XYD plus psychotherapy. Results: There was significant difference in SCL-90 scales between the healthy subjects and the FD patients before treatment (P<0.01). After treatment, the mental symptoms and the symptom of FD in the patients were markedly improved, as compared with those before treatment, the difference was significant (P<0.01). Conclusion: Mental symptoms, such as depression and anxiety existed commonly in FD patients, were closely related to FD. XYD plus psychotherapy could cure it effectively.

Objective To observe the therapeutic effectiveness and safety of autologous peripheral blood stem cell transplantation (APBSCT) for poor-risk non-Hodgkin lymphoma (NHL). Methods Ten patients with poor-prognosis NHL were enrolled from October 2003 to October 2008 in our institute. Ten patients were treated by APBSCT with CY+TBI conditioning regimen (Two patients of them were treated by Double-APBSCT with MEC conditioning regimen). Results Hematopoietic reconstitution was observed in all patients.The time of neutrophil count ≥0.5×109L and platelet≥20×109/L were at day 10 (range: 7-14) and day 16 (range: 10-37), respectively. All patients achieved complete remission (CR) after transplantation. Severe toxicity and transplant related mortality were not observed. After a median follow-up of 24 (10-84) months,seven cases were in event-free survival and three cases relapsed. One of three relapsed patients died from progression of disease, the other was still alive after treatment. Conclusion APBSCT in the treatment of patients with poor-prognosis NHL is a safe, convenient and efficient treatment.

Objective:To investigate the clinical significance of peripheral blood neutrophil to lymphocyte ratio in patients with 2 type diabetes mellitus complicated acute cerebral infarction, so as to provide theoretical basis for clinical diagnosis and treatment. Methods:From Jan. 2014 to Dec. 2016 year,136 patients with 2 type diabetes mellitus complicated acute cerebral infarction were selected as research group and 68 diabetic patients undergoing physical examination were selected as the control group. White blood cell count(WBC),neutrophil count(NC),lymphocyte count(LC) and neutrophil to lymphocyte ratio(NLR) were detected and calculated. The role of WBC,NC,LC and NLR in the diagnosis of type 2 diabetes mellitus combined with acute cerebral infarction were evaluated by receiver operating characteristic curve(ROC) and area under the curve(AUC). Results:The levels of WBC,NC and NLR in research group were significantly higher than those of in control group(P＜0. 05),while the level of LC was significantly lower than that of in control group(P＜0. 05). The levels of WBC,NC and NLR in research group increased with the increase of NIHSS score(P＜0. 05),and the level of LC decreased with the increase of NIHSS score(P＜0. 05). The area under the curve of NLR,WBC,NC and LC in diagnosing diabetes combined with acute cerebral infarction were respectively 0. 856,0. 717,0. 685 and 0. 321. The sensitivity were respectively 0. 838,0. 596,0. 517 and 0. 208. The specificity are respectively 0. 868,0. 838,0. 793 and 0. 559. Conclusion:NLR could be used as a marker for the diagnosis of type 2 diabetes with acute cerebral infarction.