Objective To establish the methods of isolating and culturing human ovarian endometriosis-derived microvascular endothelial cells (OEMEC).Methods The tissues of human endometriotic cyst of ovary were finely minced with scissors,then digested by collagenase Ⅰ,Ⅱ and trypsinethylene diamine tetraacetic acid (EDTA).The cells were purified by using centrifugation of 2000 r/min speed.OEMEC were identified by light microscope and transmission electron microscope observing CD34,FⅧ-Rag and Weibel-Palade in microvascular endothelial cells.Results The OEMEC grew as confluent monolayer like cobblestones under light microscope.CD34 and FⅧ-Rag were expressed strongly,and the percentages of CD34 and FⅧ-Rag positive cells were 91.4％ and 92.5％.Weibel-Palade bodies could be observed under transmission electron microscope.The time of cell doubling proliferation was 4.5 days.Conclusion The established system of isolating OEMEC would provide lab base for studying the mechanisms of angiogenesis in endometriosis lesions.

Objective To evaluate the role of histone deacetylase in the spinal cord in the maintenance of neuropathic pain (NP) in rats.Methods Twenty-seven male Sprague-Dawley rats,weighing 230-270 g,were randomly divided into 3 groups (n =9 each) using a random number table:sham operation group (group S),group NP,and NP + intrathecal Trichostatin A (TSA) group (group T).NP was induced by chronic constrictive injury.At 7 days after operation,5％ DMSO,5％DMSO and TSA 10 μg (10 μl) were injected intrathecally once a day for 3 consecutive days in S,NP and T groups,respectively.Mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before operation and 3,7,10,14 and 21 days after operation (T0-5).Results Compared with group S,MWT was significantly decreased and TWL was shortened at T2-5 in NP and T groups.Compared with group NP,MWT was significantly increased and TWL was prolonged at T3,4 in group T.Conclusion Histone deacetylase in the spinal cord is involved in the maintenance of neuropathic pain in rats.

Objective To evaluate the effect of intrathecal ropivacaine on spinal histone deacetylase 1 (HDAC1) and HDAC2 expression in the rats with neuropathic pain (NP).Methods Thirty adult male Sprague-Dawley rats, weighing 220-250 g, in which intrathecal catheters were successfully placed, were randomly divided into 3 groups (n=10 each) using a random number table: sham operation group (group S), group NP, and NP + ropivacaine group (group R).NP was induced by chronic constriction injury (CCI) in anesthetized rats.Sciatic nerve was exposed and 4 loose ligatures were placed on the left sciatic nerve at 1 mm intervals with 4-0 chromic catgut.Starting from 7th day after CCI, 0.25％ ropivacaine 20 μl was injected intrathecally in group R, while the equal volume of normal saline was given instead of ropivacaine in S and CCI groups once a day for 7 consecutive days.At 1 day before CCI (T0) ,and 3, 7, 10, 14, 17 and 21 days after CCI (T1-6) , the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured.After the pain threshold was measured at T4,3 rats from each group were sacrificed, and their lumbar enlargements were harvested for determination of the expression of HDAC1 and HDAC2 by Western blot.Results Compared with group S, the MWT was significantly decreased, and the TWL was shortened at T2-6, and the expression of HDAC 1 and HDAC2 was up-regulated at T4 in NP and R groups (P＜0.05).Compared with group NP, the MWT was significantly increased at T4, and the TWL was prolonged at T3.4, and the expression of HDAC1 and HDAC2 was downregulated at T4 in group R (P＜0.05).Conclusion The mechanism by which intrathecal ropivacaine alleviates NP may be related to inhibited up-regulation of spinal HDACI and HDAC2 expression in rats.

[Objective]To analyze the clinical features and related risk factors in diabetic retinopathy(DR)and diabetic periph?eral neuropathy(DPN),the two micro-peripheral vascular diseases in newly diagnosed type 2 diabetes mellitus.[Methods]A retro?spective study of 211 cases of newly diagnosed type 2 diabetes mellitus inpatient from 2009 to 2012 ,and compare the two micro-peripheral vascular complications.[Results]The morbidity of DPN was(33.6%),which was higher than that of DR,some cases of DR co-existed with DPN,DR was related with DPN(r=0.158,P=0.020). Age and systolic blood pressure were the common risk factors in DR and DPN by single factor analysis.[Conclusion]It should be paid attention to the screening of both DR and DPN in each age group in newly diagnosed type 2 diabetes mellitus. Except for the control of blood glucose ,the control of the systolic blood pressure is important in the prevention and treatment in the two micro-peripheral vascular diseases.

Objective To investigate the roles of myeloid differentiation factor 88 (MyD88) and TIR domain-containing adaptor inducing interferon-β (TRIF) in sepsis-induced myocardial dysfunction, and to analyze whether strain rate (SR) can be early sensitive evaluation for septic heart failure.Methods Sixty-four healthy male C57BL/6 mice were divided into four groups by random number table (n = 16 in each group): sham group, cecum ligation and puncture (CLP)-induced sepsis model group, anti-MyD88 group and anti-TRIF group. The anti-MyD88 group and anti-TRIF group were injected with 5μL/g of anti-MyD88 antibody or anti-TRIF antibody through the tail veins 2 hours before CLP. Eight animals in each group were used to observe the survival of 24 hours, and the other 8 myocardial tissues were harvested for examination. The cardiac function was evaluated by echocardiography before and 6 hours and 12 hours after operation. The mRNA expressions of MyD88, TRIF and inflammatory factors in myocardium were measured by polymerase chain reaction (PCR) at 24 hours after operation, and the degree of neutrophils infiltration was detected by myeloperoxidase (MPO) activity.Results The number of 24-hour survive in anti-MyD88 group and anti-TRIF group were higher than that in CLP group (number: 4, 3 vs. 2,P = 0.044,P = 0.047). Compared with sham group, the cardiac function was significantly decreased, the mRNA expressions of myocardial tissues MyD88, TRIF, interleukin (IL-1, IL-6) and tumor necrosis factor-α (TNF-α) were significantly increased, and the infiltration of neutrophils were obvious in CLP group. Compared with CLP group, the left ventricular short axis fractional shortening rate (FS) and SR were significantly increased after 12 hours in anti-MyD88 group and anti-TRIF group [FS: (49.52±1.78)%, (49.89±1.49)%vs. (41.11±1.63)%, SR (s-1): 17.63±2.16, 17.85±1.64 vs. 12.55±1.84]; the mRNA expressions of MyD88, TRIF and inflammatory factors were significantly decreased [MyD88 mRNA (A value): 0.463±0.046, 0.505±0.048 vs. 0.638±0.102, TRIF mRNA (A value): 0.413±0.031, 0.410±0.021 vs. 0.625±0.057, IL-1 mRNA (A value):0.569±0.101, 0.570±0.091 vs. 0.946±0.171, IL-6 mRNA (A value): 0.551±0.143, 0.431±0.157 vs. 0.850±0.194, TNF-α mRNA (A value): 0.471±0.082, 0.444±0.093 vs. 0.707±0.094]; and the infiltration of neutrophils were significantly decreased [MPO (U/L): 62.34±2.60, 60.87±2.40 vs. 73.83±4.90], with statistically significant differences (allP < 0.05). There was no statistical difference in above parameters between the anti-MyD88 group and anti-TRIF group (allP > 0.05).Conclusions Blocking MyD88 and TRIF expression play significant and similar roles in protecting cardiac deterioration from sepsis by attenuating cytokine release, reducing neutrophil infiltration. SR can sensitively assess septic cardiac dysfunction.

Objective To observe the effect of reciprocating gait orthosis on walking ability and activities of daily living (ADL) of patients with C6～T6 spinal cord injury (SCI).Methods 20 cases with C6～T6 SCI and according with type A and type B of ASIA standard received rehabilitation therapy, including training of muscle power, cardiopulmonary function, transferring mobility, ADL, bladder function, etc., before assembling the reciprocating gait orthosis, and additional gait training after fixing orthosis. The timed walking function assessment (6-min walking test and timed 10 meters walking test), gait analysis, Modified Barthel Index (MBI) and Functional Independence Measure (FMI) were used to assess pre- and post-training.Results The ADL ability of all cases improved obviously after equipping reciprocating ambulation orthosis, the walking ability also improved (P＜0.05). The mean timed 10 meters was 99.21±75.24 s, mean 6-min walk test was 41.71±29.38 m. From all patients, 14 cases achieved practicability walking ability, 6 cases achieved domestic walking ability.Conclusion The reciprocating gait orthosis combined with comprehensive rehabilitation therapy has effect on walking ability and ADL of patients with C6～T6 SCI.

Objective To investigate the effect of predeposit autotransfusion in operation of the patients with lumbar disc protrusion.Methods Fifty patients of transfusion with lumbar disc protrusion were assigned into two groups by stratified sampling randomly,30 patients whose blood were predeposited before operation in experimental group,and the other 20 patients whose blood were not predeposited before operation in control group.The blood loss,the blood requirements during operations,the hemotological routine indexes and the complications related to blood transfusion were compared respectively.Results The blood loss of experimental group [ (720 ± 665 ) ml ] perioperative period was lower than that of control group [ ( 1060 ± 558 ) ml ],but there was no significant difference between two groups (P ＞ 0.05 ).All the patients in experimental group went through perioperative period safely without allogenic blood transfusion.Hemoglobin,red blood cell and white blood cell were not significantly different between two groups before and after operation for 3,7 days (P＞ 0.05 ),the platelet count after operation for 7 days was significantly different between two groups (P ＜ 0.05).No complication was observed in experimental group but 1 case with complication was observed in control group.Conclusions Predeposit autotransfusion is an effective to avoid homologous blood transfusion and its complications for the patients with lumbar disc protrusion.Furthermore,the clinical effect is not significantly different between the predeposit autotransfusion patients and the allogenic blood transfusion patients.

Thirty cases of pituitary stalk interruption syndrome were divided into normal height (group 1) and short stature (group 2).There was no significant difference in growth hormone or insulin-like growth factor-Ⅰ levels between two groups(P＞0.05).Expected height and body mass index in group 1 were higher than those in group 2,while testosterone,cortisol,and FT4 were lower.Height were positively correlated with age,expected height,birth weight,and body mass index.The height of patients with pituitary stalk interruption syndrome depended on both genetic factors and the hormones from pituitary-target gland.

Aim To study the mechanism of cucurb-itacin E ( CuE )-induced autophagy in HeLa cells. Methods Improved MTT assay was adopted to meas-ure the effect of CuE on cell proliferation. Western blot was used to determine the phosphorylation levels of downstream signaling proteins of mTORC1 and the ex-pression of autophagy associated proteins. ResultsCuE inhibited the proliferation of HeLa cells in a dose-dependent manner, and the 24-h IC50 of CuE was 4. 01μmol· L-1 . CuE significantly inhibited the phospho-rylation of p70 S6 K in a time-and dose-dependent man-ner as evidenced by decreased phosphorylation levels of
the mTORC1 substrate. Meanwhile, the expression of LC3-II, a marker for autophagosome formation, was elevated by CuE treatment, and was further increased in the presence of chloroquine. Furthermore, CuE re-duced the levels of p62/SQSTM1 . These results indi-cated that CuE induced autophagy in HeLa cells. The decreased levels of phosphorylated ULK1 S757 were posi-tively correlated with autophagy induction in HeLa cells. Conclusion CuE is likely to induce autophagy through inhibiting mTORC1 activity.

The purpose of this study is to assess the effects of anti-inflammatory on activation of nuclear factor-kappaB and mRNA and protein expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in intestinal mucosal biopsy specimens from patients with ulcerative colitis (UC). A total of 27 cases with UC were investigated. 15 cases received sulfasalazine (SASP) treatment or SASP and glucocorticoid treatment, 12 cases did not receive any medication related with UC. Normal mucosa from 9 colon cancer cases served as control. Ten pieces of intestinal mucosal biopsy specimens were obtained from each patient. The mRNA expression of ICAM-1 and VCAM-1 were determined by reversal transcription-polymerase chain reaction (RT-PCR). The protein levels of ICAM-1 and VCAM-1 were measured by enzyme linked immunosorbent assay (ELISA). NF-kappaB DNA binding activity was evaluated by electrophoretic mobility shift assay (EMSA). The results showed that NF-kappaB DNA binding activity, mRNA and protein expression of ICAM-1 and VCAM-1 were increased significantly in patients with UC, compared with normal control (P<0.05). Glucocorticoids and SASP markedly inhibited NF-kappaB activation and significantly decreased mRNA and protein expression of ICAM-1 and VCAM-1 (P<0.05). Adhesion molecules (ICAM-1 and VCAM-1) gene activation had significant positive correlation with the NF-kappaB DNA binding activity (r=0.8652 P<0.05, r=0.7902, P<0.05, respectively). We concluded that NF-kappaB is a major and essential factor in regulating the expression of adhesion molecules, it plays an important role in the pathogenesis of UC. SASP and glucocorticoids ameliorate UC via inhibition of NF-kappaB activation and reduction of adhesion molecules expression.
Adult ; Anti-Inflammatory Agents ; therapeutic use ; Colitis, Ulcerative ; drug therapy ; metabolism ; Female ; Humans ; Intercellular Adhesion Molecule-1 ; biosynthesis ; genetics ; Intestinal Mucosa ; metabolism ; Male ; NF-kappa B ; metabolism ; Prednisone ; therapeutic use ; RNA, Messenger ; biosynthesis ; genetics ; Sulfasalazine ; therapeutic use ; Vascular Cell Adhesion Molecule-1 ; biosynthesis ; genetics