Objective:To investigate the impact and mechanisms of TAM to the imbalance of Treg /Th17 in the Eoc microenvi-ronment.Mte hods:Build the in vitro M2 macrophage model ,which was like TAMs .Use flow cytometry to detect the difference of the Treg/Th17 before and after the co-culture of M2 macrophage and CD 4+T cells.Use Western bolt to detect the change of T cell transcription factor and ELISA to detect the IL-10 levels in the supernatant after co-culture.Use crystal violet methods to detect the influence to the ovarian tumor cell proliferation between the different co-culture supernatants and the Transwell to detect the influence to the ovarian tumor cell migration.Thus to analysis the how TAMs influence the imbalance of Treg/Th17 in Eoc microenvironments.R esults:①After coc-ultured with M2 macrophage ,the ratio of Treg/Th17 was( 0.76 ±0.33 ) significant increased compared with control (0.41±0.25) ,M0( 0.40±0.32) and M1(0.31±0.16) (P<0.05).②After co-cultured,the supernatant of M2 group has a significant ability to promote the proliferation of Skov-3 cells.After co-cultured for 1 day, the Skov-3 cell number of M2 group was 14 942.43 ±434.19 , which was significantly higher than the control group ( 12 445.57 ±179.34 ) and CD3/28 group (12 470.32±434.18)(P<0.001).After co-cultured for 2 days,the Skov-3 cell number of M2 group was 30 129.09±520.53 ,which was significantly higher than the control group (25 622.81±897.07) and CD3/28 group(25 721.62±1 808.60) (P<0.05).③After co -cultured with M2 macrophage , the Treg-specific transcription factor Foxp 3 increased ( P=0.047 ) compared with control and M 1 group .④After co-cultured with M2 macrophage for 3 days,the concentration of IL1-0 in the supernatant was(264.04±75.9)pg/ml, which was significantly higher than CD 3/28 group ( 60.89 ±46.54 ) pg/ml,M 0 group ( 44.81 ±32.93 ) pg/ml, M1 group ( 42.71 ± 26.09)pg/ml(P=0.001).Conclusion: M2 macrophage induces the increase of the radio of Treg /Th17 as well as the increase of Treg-specific transcription factor Foxp 3 and the decrease of Th17 -specific transcription factor ROR-γt.Meanwhile , the co-culture supernatant of M2 macrophage and CD4+T cell have the ability to promote the proliferation and migration of ovarian cancer cell ,the mechanism which ,may related to the IL -10 in the supernatant .

Objective To investigate the memory monitoring ability in patients with idiopathic generalized epilepsy (IGE) and explore the mechanism of the memory impairment.Methods The feeling of knowing (FOK) paradigm of episodic memory (EM) and semantic memory (SM) were established and subsequently administered in 31 patients with IGE (IGE group) and 30 healthy controls (HC group) participants who were matched in age,sex and educational level.Results Compared with HC group (feeling of knowing of episodic memory (FOK-EM) FOK accuracy (85.29± 16.84) ％;feeling of knowing of sematic memory (FOK-SM) recall (76.61± 18.66) ％),the FOK-EM FOK accuracy ((64.03± 22.10) ％) and FOK-SM recall ((53.27±26.91) ％) in IGE group were significantly decreased(t=-4.215,P＜0.01;t=-3.677,P＜0.01).The correct judgment and correct recognition of FOK-EM ((51.16±32.93) ％) and the false judgment and correct recognition ((21.07±24.38) ％) of FOK-EM in the IGE group were significantly different with the HC group (the correct judgment and correct recognition of FOK-EM:(79.34±27.26)％ and the false judgment and correct recognition of FOK-EM:(2.45±5.38) ％;t=-3.634,P＜0.01;t=4.149,P＜0.01).Most importantly,the false judgment and correct recognition of FOK-EM were correlated with the Digital Span Test,Vocabulary Fluency Test and the Stroop effect in IGE group (r=-0.309,P＜0.05;r=-0.355,P＜0.01;r=-0.354,P＜0.05;r=0.602,P＜0.01).Conclusion The results show that the IGE group made less accurate metamemory monitoring than the HC group by underestimating their memoU performance on FOK-EM,whereas the semantic metamemory monitoring is not impaired in IGE group.More importantly,the impairment of memory monitoring was correlated with the deficit of executive function,indicating that this mechanism can be an influential factor of memory disorder in IGE.It also indicates that the episodic and semantic metamemory monitoring depend on different neural networks.

OBJECTIVETo determine the expression of Skp2 in different prostate cancer (PCa) cell lines and tissues, and explore its influence on the androgen receptor (AR) signaling pathway and development of castration-resistant prostate cancer (CRPC).

METHODSThe expression levels of Skp2 and AR in different PCa cell lines were detected by Western blot. After knockdown of Skp2 in the C4-2 and 22RV1 cells transfected with shRNA, the expressions of AR and P27 were determined and the activity of ARR3-Luc measured by dual-luciferase reporter gene assay following treatment with dihydrotestosterone (DHT). The expressions of AR and Skp2 in human naïve PCa or CRPC specimens were detected by immunohistochemical staining followed by analysis of their differences and correlation.

RESULTSThe Skp2 protein expression level was significantly higher in the C4-2 or 22RV1 cells than in the LNCaP cells. DHT treatment increased the expression of Skp2 in the C4-2 cells, but knock-down of Skp2 significantly up-regulated the expression of the well-known downstream protein P27 and down-regulated that of AR. Consistently, DHT treatment increased the activity of ARR3-Luc, while knockdown of Skp2 remarkably decreased it in the C4-2 and 22RV1 cells (P < 0.05). In addition, significantly higher expressions of Skp2 and AR were observed in the CRPC than in the naïve specimens (P < 0.05), with a positive correlation between the two proteins (r = 0.658 1, P < 0.05).

CONCLUSIONSkp2 can enhance the expression and transcription activity of the AR protein in CRPC cells or tissues and is promising to be a critical molecular therapeutic target.

Androgens ; pharmacology ; Cell Line, Tumor ; Dihydrotestosterone ; pharmacology ; Disease Progression ; Gene Knockdown Techniques ; Humans ; Male ; Neoplasm Proteins ; genetics ; metabolism ; Prostatic Neoplasms, Castration-Resistant ; metabolism ; Receptors, Androgen ; genetics ; metabolism ; S-Phase Kinase-Associated Proteins ; physiology ; Transcriptional Activation ; Up-Regulation

BACKGROUNDVanishing white matter disease (VWM), a human autosomal recessive inherited leukoencephalopathy, is due to mutations in eukaryotic initiation factor 2B (eIF2B). eIF2B is responsible for the initiation of protein synthesis by its guanine nucleotide exchange factor (GEF) activity. Mutations of eIF2B impair GEF activity at different degree. Previous studies implied improperly activated unfolded protein response (UPR) and endoplasmic reticulum stress (ERS) participated in the pathogenesis of VWM. Autophagy relieves endoplasmic reticulum load by eliminating the unfolded protein. It is still unknown the effects of genotypes on the pathogenesis. In this work, UPR and autophagy flux were analyzed with different mutational types.

METHODSERS tolerance, reflected by apoptosis and cell viability, was detected in human oligodendrocyte cell line transfected with the wild type, or different mutations of p. Arg113His, p. Arg269FNx01 or p. Ser610-Asp613del in eIF2Bε. A representative UPR-PERK component of activating transcription factor 4 (ATF4) was measured under the basal condition and ERS induction. Autophagy was analyzed the flux in the presence of lysosomal inhibitors.

RESULTSThe degree of ERS tolerance varied in different genotypes. The truncated or deletion mutant showed prominent apoptosis cell viability declination after ERS induction. The most seriously damaged GEF activity of p. Arg269FNx01 group underwent spontaneous apoptosis. The truncated or deletion mutant showed elevated ATF4 under basal as well as ERS condition. Decreased expression of LC3-I and LC3-II in the mutants reflected an impaired autophagy flux, which was more obvious in the truncated or deletion mutants after ERS induction.

CONCLUSIONSGEF activities in different genotypes could influence the cell ERS tolerance as well as compensatory pathways of UPR and autophagy. Oligodendrocytes with truncated or deletion mutants showed less tolerable to ERS.

Cell Line ; Endoplasmic Reticulum Stress ; genetics ; physiology ; Eukaryotic Initiation Factor-2B ; genetics ; Humans ; Mutation ; genetics ; Oligodendroglia ; metabolism ; Unfolded Protein Response ; genetics ; physiology

OBJECTIVETo compare the effects of two contrast agents, Gd-DTPA and HSA-Gd-DTPA, in magnetic resonance (MR) lymphography.

METHODSTwelve New-Zealand rabbits were randomized into Gd-DTPA and HSA-Gd-DTPA groups with subcutaneous (interdigital skin fold) injection of the two contrast agents (0.2 ml of 0.5 mmol/L Gd(3+)) for MR lymphography of the popliteal lymph nodes examined in the axial and sagital orientation. T(1)-weighted, T1-weighted fat suppressed, and T(2)-weighted spin-echo (SE) images of the lymph nodes were obtained in plain scans. The post-contrast scanning started at 30 min, 1 h and 3 h after Gd-DTPA administration and at 10 min, 30 min and 60 min after HSA-Gd-DTPA injection to obtain T(1)-weighted images with identical imaging parameters. The signal intensity of popliteal lymph node was measured and the enhancement rate calculated.

RESULTSAfter subcutaneous injection, Gd-DTPA quickly entered blood circulation to result in obvious enhancement of the anterior-tibial vein and the urine and also in heterogeneous enhancement of the popliteal lymph nodes. HSA-Gd-DTPA did not enter the blood, causing obvious homogeneous enhancement of the lymphatic vessels and lymph nodes. HSA-Gd-DTPA resulted in higher enhancement rate than Gd-DTPA, and the enhancement rate in Gd-DTPA group decreased with time as opposed to that of the HSA-Gd-DTPA group.

CONCLUSIONHSA-Gd-DTPA has better performance than Gd-DTPA in MR lymphography after subcutaneous administration.

Animals ; Contrast Media ; administration & dosage ; pharmacokinetics ; Gadolinium DTPA ; administration & dosage ; pharmacokinetics ; Humans ; Lymph Nodes ; diagnostic imaging ; Lymphography ; instrumentation ; methods ; Rabbits ; Random Allocation ; Serum Albumin ; administration & dosage ; pharmacokinetics

Objective:To preliminarily verify the cross talk between tissue factor/active coagulation factor Ⅶ (TF/FⅦa) and epidermal growth factor receptor (EGFR) pathways in human colon cancer cells in culture.Methods:FⅦa was treated to HT-29 (KRAS-wild type) and LoVo (KRAS-mutant) colon cancer cells to activate TF/F Ⅶa pathway,qRT-PCR and Western blot were used to detect the expressions of amphiregulin (AREG) and epiregulin (EREG),ligands of EGFR on mRNA and protein levels,respectively.After knocking down expression of TF by TF-targeted siRNA transfection,FⅦa was treated and mRNA expressions of AREG and EREG were detected to see whether the FⅦa-induced effects were dependent on TF.Expressions of mRNA of TF and FⅦwere detected by qRT-PCR following the activation of EGFR pathway by treatment with epidermal growth factor (EGF) to HT-29 and LoVo cells.Results:After TF/FⅦa pathway was activated,for HT-29 cells,expressions of AREG (on mRNA level) and EREG (both on mRNA and protein level) were significantly down-regulated versus those of control group,gene expressions of AREG and EREG were 0.55 ± 0.09 vs.0.99 ± 0.09,0.67 ± 0.10 vs.1.02 ± 0.02,protein expressions of EREG were 0.54 ± 0.09 vs.1.04 ± 0.13,all P ＜ 0.05.For LoVo cells,expressions of AREG (both on mRNA and protein level) and EREG (on protein level) were significantly up-regulated versus those of control group,gene expression of AREG were 1.87 ± 0.39 vs.0.93 ± 0.23,protein expressions of AREG and EREG were 3.09 ±0.73 vs.1.11 ±0.21,1.53 ±0.19 vs.0.97 ± 0.23,all P ＜0.05.The regulating effect of AREG and EREG mRNA expression by FⅦa in HT-29 and LoVo cells could both be partly blocked by knocking down TF expression.For HT-29 cells,activation of EGFR pathway induced no significant TF mRNA expression,F Ⅶ mRNA expression was not detected.However,for LoVo cells,activation of EGFR pathway induced significantly higher mRNA expressions of both TF and FⅦ,expressions were 1.53 ± 0.23 vs.1.00 ± 0.23,53.20 ± 6.08 vs.1.00 ± 0.15,all P ＜0.05.Conclusion:In colon cancer cell LoVo,when activated,TF/FⅦa pathway and EGFR pathway could interact through upregulating the other pathway's effectors,and mutant KRAS might play a critical role in the two pathways'cross talk.

OBJECTIVETo study the risk factors of secondary kyphotic angle increment after bone cement vertebroplasty for osteoporotic vertebral compression fractures.

METHODSFrom October 2005 to May 2006, 32 (45 vertebrae) bone cement vertebroplasty procedures were performed. The operation time, injected cement volume, bone mineral density, visual analog scale (VAS) pain score, vertebral height, and kyphotic angle were recorded. The secondary increment of the kyphotic angle was calculated, and correlation analysis and linear regression analysis were performed.

RESULTSThe bone mineral density, the postoperative kyphotic angle and the vertebral midline height were significantly correlated to the secondary increment of the kyphotic angle.

CONCLUSIONLarge postoperative kyphotic angle, poor postoperative recovery of the vertebral midline height, and low bone mineral density are all risk factors of secondary increment of the kyphotic angle.

Aged ; Aged, 80 and over ; Female ; Fractures, Compression ; etiology ; surgery ; Humans ; Kyphosis ; etiology ; pathology ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Osteoporosis ; complications ; surgery ; Risk Factors ; Spinal Fractures ; etiology ; surgery ; Thoracic Vertebrae ; surgery ; Treatment Outcome ; Vertebroplasty ; adverse effects

OBJECTIVETo investigate how to select an appropriate surgical approach in the management of fracture and dislocation of lower cervical spine combined spinal cord injury.

METHODSThe clinical data of 54 patients of lower cervical spine fracture and dislocation were retrospectively analyzed. There were 29 cases with vertebral body compressive fracture and dislocation, 7 cases with vertebral body bursting fracture and dislocation, 3 cases with unilateral facet dislocation, 15 cases with bilateral facet dislocation. All cases were associated with spinal cord injury. According to American Spinal Injury Association (ASIA) grades, 21 cases were in A grade, 5 cases in B grade, 22 cases in C grade and 6 cases in D grade. All patients had surgical reduction, decompression, stabilization and fusion, 43 cases in anterior approach and 11 cases in posterior approach.

RESULTSAll patients were followed up in 12 to 36 months, the mean follow-up time was 18 months. There were no great vessels, trachea, esophagus or spinal cord iatrogenic injury. There were no pull-out and breakage of screws or plates. Fusion was achieved in all patients at an average of 12 weeks postoperatively. There were no pseudarthrosis or bone nonunion. Of all the patients, 96.3% were acquired completely reduction and the normal intervertebral height and lordosis were maintained. Patients with complete spinal cord had no neurologic recovery, but they felt relief from upper limb pain or numb. Incomplete spinal cord lesions improved on average 1-2 Frankel grade after surgery.

CONCLUSIONSFor lower cervical spine fracture and dislocation, an ideal anatomy reduction can be obtained with either anterior or posterior approach surgery. It is important to select a suitable surgical approach according to different types of cervical fracture and dislocation.

Adolescent ; Adult ; Aged ; Cervical Vertebrae ; injuries ; surgery ; Diskectomy ; methods ; Female ; Humans ; Joint Dislocations ; complications ; surgery ; Laminectomy ; methods ; Male ; Middle Aged ; Retrospective Studies ; Spinal Cord Injuries ; complications ; Spinal Fractures ; complications ; surgery ; Spinal Fusion ; methods ; Treatment Outcome

OBJECTIVETo research the optimal conditions for the callus induction of anther culture and the plant regeneration of Angelica dahurica var. formosana.

METHODCallus was induced from the anther of A. dahurica from Sichuan province on a MS medium. The effects of callus induction and plant regeneration of different pretreatment hours under low temperature (4 degrees C), different culturing conditions under darkness and illumination, and different culture with different hormone contents and ratios were studied.

RESULTThe results showed that A. dahurica anthers without low temperature pretreatment reached the highest induction rate then under the pretreatment under low temperature (4 degrees C) for two days. The optimal culturing condition was under the darkness. The culturing efficiency reached 38.89% on the medium of MS + 2.0 mg x L(-1) 2,4-D + 1.0 mg x L(-1) 6-BA. The optimum medium for differentiate anther callus was MS + 0.5 mg x L(-1) NAA + 1.5 mg x L(-1) KT + 10 mg x L(-1) AgNO3. 1/2MS medium supplemented with 0.5 mg x L(-1) IBA could well promote seedings to take roots.

CONCLUSIONAn efficient system for callus induction of anther culture and plant regeneration of A. dahurica was preliminarily established.

Angelica ; drug effects ; growth & development ; Culture Media ; chemistry ; pharmacology ; Flowers ; Tissue Culture Techniques ; methods

Objective To evaluate the clinical efficacy and safety of endoscopic selective varices devascularization.Methods From November 2015 to July 2017,at the First Affiliated Hospital of Anhui Medical University,282 patients with liver cirrhosis complicated with gastroesophageal varices who underwent endoscopic treatment were selected and divided into traditional treatment group (n=102) and selective treatment group (n=180).The patients of traditional treatment group were treated with tissue glue "sandwich method" (lipiodol-tissue glue-lipiodol),and the patients of selective treatment group were treated with modified " sandwich method" (lauromacrogol-tissue glue-0.9％ sodium chloride solution).After operation,all the patients were followed up for three months.The rates of remarkable efficacy,efficacy,improvement,rebleeding and complications were compared between the two groups respectively.T test,chi-square test and rank sum test were performed for groups comparison.Results One month after operation,the rates of remarkable efficacy and efficacy of the traditional treatment group and the selective treatment group were 46.1％ (47/102) and 20.6％ (21/102),and 67.8％ (122/180) and 30.0％ (54/180),respectively.The results of rank sum test indicated that the differences in rates of remarkable efficacy and efficacy between two groups were statistically significant (Z=-5.428,bilateral P＜0.05).The improvement rate of the selective treatment group was higher than that of the traditional treatment group (97.8％,176/180 vs.66.7％,68/102),and the difference was statistically significant (x2-54.048,P＜ 0.05).At two weeks and three months after operation,the rebleeding rate of traditional treatment group was higher than that of selective treatment group (10.8 ％,11/102 vs.3.3 ％,6/180;21.6％,22/102 vs.7.2％,13/180),and the difference was statistically significant (x2 =6.380 and 12.327,both P＜0.05).No serious complications occurred in both groups.There was no statistically significant difference in overall complication rate between selective treatment group and traditional treatment group (37.8％,68/180 vs.30.4％,31/102;x2 =1.559,P-0.212).Conclusion The treatment with endoscopic selective varices devascularization shows good efficacy and safety,and is worth further study.