Objective To study the mechanism of hypoxia inducing factor-1α(HIF-1α)pathway in establishment of hypoxia inducing low endometrial receptivity.Methods RL95-2 cell lines.the ideal model of study ER,were cultured in hypoxia condition induced by CoCl2,and the expression of mRNA and protein of HIF-1α and tumor necrosis factor like weak inducer of apoptosis(TWEAK)were measured by reverse transcription-PCR and western blot. The apoptosis rate was analyzed by flow eytometry.Then the mechanism confirmed by comparing the two factors in endometrium and the ultra-appearance of inflammatory reaction and apoptosis between recurrent spontaneous abortion women and control women.Results (1)On difierent time point(0,12,24,48 hour),mRNA expression of HIF-1α were 0.272±0.010,0.354±0.020,0.591±0.020.0.890±0.020,while the expression of TWEAK were 0.104±0.010,0.510±0.020,1.021±0. 020, 1. 237 +0. 040, respectively, the expression level between 12, 24, 48 and 0 hour all showed significant differences (P＜0. 05 ). (2) Protein expression of HIF-1α were 0. 853 +0. 010, 0. 931 ±0. 030,1. 124±0.010, 1.317±0.0 20 respectively, while was 0.042±0.010, 0.091 ±0.010, 0. 131±0.020,0. 205 ±0. 030 in TWEAK expression, the different level were statistically significant ( P＜0. 05 ). ( 3 )With longer culture under hypoxia, the cell apoptosis rate increased obviously. The apoptosis rate of each time point were ( 3.2±1.4 ) %, ( 16. 2 ±3.2 ) %, ( 26. 3±3.5 ) %, ( 31.8±3.5 )%, the differences between 12, 24, 48 and 0 hour had significance (P ＜0. 05). (4) The positive rate of HIF-1α stained in epithelium cells and stroma cells of test group were 32. 3%, 8.4% and 16. 7%, 7. 3% in control group. The positive rate of TWEAK were 28. 3%, 3.9% in recurrent spontaneous abortion group and 11.6%, 2. 7% in control group ( P ＜0. 05 ). The ultra-appearance of inflammatory cell infiltrated and apoptosis were obvious in test group. Conclusions Cell inflammation reaction and apoptosis induced by HIF-1α pathway may participate the mechanism of hypoxia inducing low endometrial receptivity. HIF-1α might become a novel target for improving poor endometrial receptivity.

Adolescent ; Adult ; Ankylosis ; complications ; surgery ; Female ; Humans ; Imaging, Three-Dimensional ; Male ; Micrognathism ; complications ; surgery ; Models, Anatomic ; Skull ; anatomy & histology ; Temporomandibular Joint Disorders ; complications ; surgery ; Young Adult

Objective To investigate the brain magnetic resonance imaging (MRI) features and clinical manifesta-tions of the patients with citrullinemia, and to promote awareness of, early diagnosis of and better treatment for the disease.Methods One case with citrullinemia was reported, and other eight cases reported in the literature in nearly 14 years were reviewed.Results The case was a 15-month-old girl with type I citrullinemia diagnosed by the mutation analysis of the ASS1 gene performed in local hospital after birth. The patient was admitted to our hospital for recurrent lethargy for a year and the high level of blood ammonia (311 μmol/L, normal range 9-33 μmol/L). The blood ammonia reduced to normal on the 11th day after arginine treatment. On MRI scans, the diffusion weighted imaging (DWI) showed diffuse hyperintensity on bilateral frontal, parietal and temporal cortex, which indicated restricted diffusion due to cytotoxic edema. On the follow-up MRI after 10 day's treatment, the affected regions was similar but the intensity decreased compared to the previous scan,which accompanied by cere-bral atrophy. Eight cases in the literature were reviewed, and the clinical manifestations in these patients were lack of speciifcity, the most common features included feeding dififculties, lethargy, and vomiting. Brain MRI was performed on 7 cases, computed tomography (CT) was performed on 1 case, with the result of cytotoxic edema in 3 cases and atrophy in 2 cases.Conclusions Citrullinemia often lacks of speciifc symptoms in the early phase. Brain MRI could provide the clinician a valuable help for early diagnosis and treatment of this disease.

Objective To explore the impact of dexamethasone on inflammatory response of thyrocytes.Methods Primary thyrocytes were extracted from thyroid tissue of patients with Graves' disease.The cells were stimulated with interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α),and cultured in dexamethasone.Thyrocytes were divided into 4 groups:control group,dexamethasone group,TNF-α + IFN-γ group,and dexamethasone+TNF-α+IFN-γ group.Interferon-γ-induced protein 10 (CXCL10) and CCL2 in supernatant of cell cultured in 4 groups were detected by enzyme-linked immunosorbent assay.Cell protein in 4 groups was extracted and GSK-3β,P50,and P100 protein were detected by Western blotting.Results MTT assay demonstrated that 10-5 mmol/L concentration of dexamethasone was optimal for cell culture.The CXCL10 level in TNF-α+IFN-γ group was higher than that in the control group and dexamethasone group (P＜0.01),but no difference was found between dexamethasone+TNF-α+IFN-γgroup and TNF-α+IFN-γgroup(P＞0.05).The CCL2 level in TNF-α+IFN-γ group was higher than that in control group and dexamethasone group(P＜0.01).There was a significant lowering of CCL2 level in dexamethasone + TNF-α + IFN-γgroup compared with TNF-α + IFN-γ group (P ＜ 0.05).The expression of GSK-3β and P100 protein was increased in TNF-α + IFN-γgroup compared with control group.The expression of GSK-3β and P100 protein was lower in dexamethasone+TNF-α + IFN-γ group than that in TNF-α + IFN-γ group.Conclusion TNF-α + IFN-γ could stimulate the secretion of CXCL10 and CCL2 in thyrocytes and thus activate the inflammatory response.Dexamethasone could reduce CCL2 secretion.Dexamethasone had little effect on CXCL10.Dexamethasone could reduce GSK-3β and P100 expressions,and inhibit the activation of NF-κB signaling pathway and thus the inflammatory response.

Objective To study the relationship between reactive oxygen species (ROS) as well as total antioxidant capacity ( TAC ) within follicle fluid and body mass index ( BMI ) in patients with polycystic ovary syndrome (PCOS).Methods All patients enrolled in this study were infertile women receiving in vitro fertilization-embryo transfer (IVF-ET) treatment.55 PCOS patients were divided into over-weight group ( n =23 ) and non-over-weight group ( n =32 ).Another 55 age-matched non-PCOS women were also divided into control group ( n =30) and overweight group ( n =25 ).Plasma sex hormone,triglycerides,and total cholesterol were determined.On oocyte retrieval day after ovarian stimulation,ROS and TAC in follicular fluid were assayed.Results Subjects in over-weight and PCOS over-weight groups had higher triglycerides than those in control and PCOS non-over-weight groups [ ( 1.9 ±1.1,1.7 ± 0.9,1.0 ± 0.5,1.2 ± 0.7 ) mmol/L,respectively,all P＜0.05],so did total cholesterol [ ( 4.8 ± 1.2,5.2 ± 1.1,4.0 ± 0.6) mmol/L,respectively,all P＜0.05].In PCOS over-weight group,ROS and ROS/TAC within follicular fluid were ( 35.4 ± 6.7 ) RLU/S and 39.8 ± 22.0,both were higher than those in the other 3 groups ( all P＜0.05).TAC [ (0.8 ± 0.5 ) Mm] was lower in PCOS over-weight group than that among the other 3 groups( all P＜0.05 ).ROS/TAC was higher in PCOS non-over-weight group than that of control group ( 26.5 ± 14.5 vs 14.2 ± 12.5,P＜0.05 ).Univariate analysis showed that both ROS and ROS/TAC within follicular fluid in PCOS patients were positively correlated with BMI ( r =0.34 and r =0.32,both P＜0.05 ).Conclusion Abnormal oxidative stress exists in follicular fluid of PCOS patients,and the oxidative stress parameters show positive correlation with BMI.

Objective To observe the distribution of bone marrow mesenchymal stem cells(MSCs) and the effects on expression of apoptosis relative proteins Caspase 3 and Bcl-2 after intravenous transplanted into ischemic rat brains.Methods MSCs from SD rats were cultivated and proliferated in vitro and marked with CFSE.MSCs were then intravenously transplanted into middle cerebral artery occlusion(MCAO)models of SD rats.The rats were killed at different time points to observe the distribution of MSCs under fluorescence microscoDe as well as the effects on expression of apoptosis relative proteins Caspase 3 and Bcl-2 using immunohistochemical method.Results Density of Caspase 3 in immunohistochemically positive area in transplantion group were(2.81±0.35)%,(3.98±0.67)%,(5.58±0.92)%,(3.51±0.63)%,(1.64±0.29)%in 6,12,24,72 hours and in 7 days,respectively,and decreased significantly compared with those of control group[(3.92±0.44)%,(5.23±0.30)%,(6.89±0.57)%,(4.39±0.57)%,(2.29±0.21)%],the difference being significant(t=4.37,3.34,2.60,2.32,3.90,P＜0.05 or＜0.01).The density of Bcl-2 in immunohistochemically positive area in transplantation group were(4.70±0.16)%,(5.61±0.26)%,(3.00±0.28)%respectively in 6,12 hours and in 7 days,which had improved significantly compared with those of control group[(3.28±0.27)%,(4.54±0.59)%,(2.15±0.62)%],the difference being significant(t=8.32,3.25,2.54,P＜0.05 or＜0.01).Conclusions Bone marrow MSCs can exert protective effects on brain ischemia and reperfusion injury possibly by down-regulating Caspase 3 and up-regulating Bcl-2.

To observe the effect of calycosin on the proliferation and activation of primary hepatic stellate cells (HSCs) in rats, and prove calycosin shows the effects through peroxisome proliferator-activated receptor γ(PPARγ) and farnesoid X receptor (FXR). The results indicated that calycosin could inhibit HSC proliferation and expressions of activation marker smooth muscle actin-α and type I collagen. With the increase in HSC activation time, FXR expression reduced, but with no notable impact from calycosin. Calycosin could up-regulate PPARγ expression and its nuclear transition in a concentration-dependent manner. Its prohibitory effect on HSC activation could be blocked by PPARγ antagonist. In conclusion, calycosin could inhibit HSC activation and proliferation, which may be related with the up-regulation of PPARγ signal pathway.
Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Hepatic Stellate Cells ; cytology ; drug effects ; metabolism ; Isoflavones ; pharmacology ; Male ; PPAR gamma ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Cytoplasmic and Nuclear ; genetics ; metabolism ; Up-Regulation ; drug effects

OBJCETIVETo investigate the method of separation of culture of bone microvascular endothelial cells (BMECs) of human femoral head in vitro.

METHODSFrom October 2013 to January 2014,15 femoral heads without pathologic change from patients resected during hip replacement were selected involving 2 males and 13 females with a mean age of 71.2 years old ranging from 38 to 92. Cancellous bone in femoral head was bited into broken bone grain and transfered into medium in aseptic contidion. Cells were isolated by the methods of enzymic digestion and density gradient centrifugation,purified by differiential attachment. The characteristics of cells was observed by inverted microscope. vWF and CD31 immunofluorescence analysis was applied for identification of cells.

RESULTSThe number of cells was positively correlated with patients' age after 24 hours in primary culture. The older patients had the less cells numbered. After 4 to 5 days' culture, primary cells appeared short spindle,polygon shaped and cobblestone-like morphology. After 7 to 10 days' culture, primary cells proliferated densely, became fusion, arranged in swirl, and contact inhibition appeared significantly. Immunofluorescence staining revealed the cells were 100% positive for vWF and CD31, and it showed that the cultured cells were BMECs.

CONCLUSIONIt was a simple, steady, effective method with good reproducibility, by which highly purified human BMECs can be obtained.

Adult ; Aged ; Aged, 80 and over ; Cell Culture Techniques ; Cell Proliferation ; Cell Separation ; methods ; Cells, Cultured ; Endothelial Cells ; cytology ; Female ; Femur Head ; blood supply ; Humans ; Male ; Microvessels ; cytology ; Middle Aged

Objective To investigate the effect of hypert riglyceridemia on vascular endothelial function. Methods With high-resolution ultrasound, flow and nitroglycerin-induced dilatation of the brachial artery were determined in thirty hypertriglyceridemic patients and thirty healthy subjects as controls. Serum lipid and plasma endothelin (ET) were determined. Results In patients with hypertriglyceridemia,flow-induced vasodilatation was much reduced compared with that in the control subjects[(2.7±2.0)% vs (15.0±8.0)%, P<0.001］.However, vasodilatation in response to nitroglycerin were similar in both groups［(15.0±5.0)% vs (16.8±9.0)%, P>0.05].Plasma ET level in the hypertriglyceridemic group was significantly higher than that in the control group［(106.22±19.16) μg/L vs (72.37±14.06) μg/L, P<0.001].ConclusionEndothelium-dependent vasodilatation was impaired in patients with hypertriglyceridemia.