BACKGROUND: Human embryonic stem cells (hESCs) are pluripotent cells which may differentiate into tissues of all three germ layers. Such research as the feeder-free growth of hESCs is few in China. Fibroblast growth factor (FGF) is a major factor to maintain the undifferentiated state of hESCs.OBJECTIVE: To evaluate the ability of FGF at different concentrations in maintaining the undifferentiated state and pluripotency of hESC lines in the long-term culture.METHODS: Two cell lines of hES-8 and hES-18 were cultured with mouse embryonic fibroblast condition medium for 3 passages and then transferred into mouse embryonic fibroblast condition medium containing different concentrations of FGF: 100, 160, 250 μg/L for 8 passages. The hESCs were removed from the petri dish, cell clusters were digested with collagenase IV and gathered. Cell differentiation and pluripotency were observed. The eighth generation of the hESCs were collected and incubated into severe combined immunodeficiency mice, so as to observe teratoma formation. Morphologies of the cells were evaluated. Alkaline phosphatase staining, surface labeling immunocytochemical analysis and RT-PCR assay method were utilized to determine the OCT-4 expression and tumorigenesis in vivo.RESULTS AND CONCLUSION: Cultured in mouse embryonic fibroblast condition medium containing 160 and 250 μg/L FGF, two cell lines of hESC could maintain undifferentiated state: Clones were round with a high ratio of nucleus to cytoplasm. Large areas in the center of clones were undifferentiated cells, while surrounding the clones were differentiated cells; Strong positive expression for alkaline phosphatase staining was observed; Two cell lines showed high levels of OCT-4 transcription factor protein; The surface markers SSEA-4, TRA-1-60, TRA-1-81 were all positive on both two lines; The hESC clusters could form embryoid body in vivo 10 days later; 3 germ layers of teratomas were also obtained after implanted into severe combined immunodeficiency mice. Mouse embryonic fibroblast condition medium containing 100 μg/L FGF was not sufficient to maintain the long-term proliferation of hESCs, and most of the cells differentiated and died after 4 passages. Alone with concentration 160 μg/LbFGF or more could maintain two hESC lines undifferentiated stably in vitro, has no influence on the differentiation and totipotency of two cell lines.

Based on a review of literature and the current reform of public hospital corporate governance,such corporate governance is divided intocombined regulation and operation,anddivided regulation and operationmodels.Then the principal-agent theory was called into play to analyze the main problems and causes of the existing models.From the perspectives of government supervision and assessment,professionalization of hospital directors,market competition mechanism,thoughts were made on how to improve such corporate governance.

Objective:To study effects of psychological intervention and sedativn on stress response to gastroscopy.Methods:86 patients for gastroscopy were randomly divided into three groups:29 patients in group of psychological intervention watched video materials about psychological intervention before gastroscopy;29 patients in sedative group were given intravenous midazolam and propofol.; 28 patients in control group were no psychological intervention or sedation. Patient's discomfort and changes of blood pressure(BP),heart rate(HR) , saturation of pulse oxygen(SpO 2) were recorded. The level of norepinephrine(NE), adnephrin(E), dopamine(DA) and cortisol(CS)in serum were detected.Results:The incidence of cough,nausea and vomiting, and restlessness in group of psychological intervention and in sedative group was lower as compared with the control group. BP,HR,NE,E and CS in control group rose significantly during the procedure while that of patients in group of psychological intervention and in sedative group was no significant change.Conclusion:The psychological intervention or sedation with midazolam and propofol can reduce stress response to gastroscopy.

Objective To explore the effects of inflammatory mediator blocker AG490 on neurological deficits,apoptosis and expression of caspase-3 following cerebral ischemia-reperfusion(I/R) in rats.Methods The male SD rats were randomly divided into the groups sham-operation,I/R,saline and AG490;and the focal cerebral I/R models were made by middle cerebral artery thread embolism method.AG490(1 mg/kg) was intraperitoneal injection in AG490 group immediate and 12 h after ischemia-reperfusion respectively.The neurological deficits score was evaluated in 24 h after I/R in each group.The number of apoptosis in cerebral tissue was examined by d-utp nick end labeling staining(TUNEL).The expression of P-JAK2,P-STAT3,caspase-3 were detected by Western Blot.Results Compared with the groups I/R and saline,the neurological deficits score in AG490 group was significantly decreased(all P

Objective To investigate the effect of GABAB receptor in spinal cord by detecting semi-quantitatively the mRNA for isoforms of GABAB receptor in a chronic neuropathic pain model. Methods Twenty-four male SD rats weighing 200-250g were anesthetized with intraperitoneal pentobarbital. Lamina of L5 vertebra and superior articular process of L6 were removed and posterior root of left L5 spinal nerve was exposed and tightly ligated with 7/0 atraumatic suture. The animals were randomly divided into 4 groups of 6 animals in each group: group A in which animals were observed for 7 days after sham operation; group B in which animals were observed for 7 days after ligation of posterior root of L5 spinal nerve; in group C animals were observed for 14 days after sham operation and in group D animals were observed for 14 days after ligation. Pain threshold was measured by heat stimulation of bilateral hindlimbs before and on the 1st , 3rd , 7th , 10th and 14th day after operation. Lumbar spinal cord was removed on the 7th or 14th day after operation and mRNA for isoforms of GABAB receptor in the spinal cord was detected semi-quantitatively by digoxin hybridization in situ.Results Pain threshold of left hindlimb (ligated side) was significantly lower than that of right hindlimb on the 7th, 10th and 14th day after operation as measured by heat stimulation. There were less mRNA for three isoforms of GABAB receptor in group B and D as compared with group A and C respectively. Conclusion The gene expression of three isoforms of GABAB receptor in the spinal cord has plasticity and is reduced during chronic neuropathic pain.

Objective To study the transfer of ropivacaine across the single cotyledon of the term human placenta and the effects of maternal hypoproteinemia and fetal acidemia on the transfer. Methods Eighteen placentas were obtained from healthy full term parturients within 5 min after vaginal or cesarean section delivery. The dual perfused human placental model was made. The placentas were randomly divided into three groups of 6 placentas : (A) control group in which 100% fresh frozen plasma was used in both maternal and fetal circulation with pH maintained at 7.4 on both sides; (B) fetal academia group in which 100% fresh frozen plasma was used in both circulations but fetal pH was reduced to 7.0; (C) maternal hypoproteinemia group in which 50% fresh frozen plasma used in maternal circulation and 100% fresh frozen plasma in fetal circulation, pH was maintained at 7.4 on both sides. Samples were taken from the perfusate in the reservoir at 15, 30, 60, 90, 120 min after ropivacaine (2?g?ml-1) and antipyrine (10 ?g?ml-1 ) were added in maternal circulation for determination of concentrations of ropivacaine, antipyrine, glucose and lactate. Glucose consumption rate, lactate generation rate and relative and absolute transfer ratio of ropivacaine were calculated. Results Absolute transfer ratio of ropivacaine was gradually increasing with perfusion time, reaching 8.7?1.0% (A) , 10.5 ?1.6% (B) and 11.8?1.1% respectively at 120 min. Relative transfer ratio of ropivacaine was relatively constant during 120 min perfusion and was significantly higher at each time point in group B and C than in group A ( P

Constant infusion, exponentially declining infusion and program infusion of theophlline were administered to rabbits byadopting the rabbit population pharmacokinetic parameters in this laboratory, and the plasma concentrations of the drug was measured by ul- traviolet spectrophotometry. The results showed that the plasma drug concentrations following exponentially declining infusion and program infusion attained the desired steady-state level at only 30 min, though the T1/2?= 6. 08h, The median absolute value of performance error in pro-gram infusion and exponentially declining infusion was 7. 8% and 14% respectively.

Objective To investigate the clinical and characteristic imaging findings of intracranial gliosarcoma. Methods The imaging findings of surgery and pathology proved intracranial gliosarcoma in 15 cases were retrospectively analyzed. There were 10 males and 5 females, ranging in age from 44 to 67 years (mean age 58 years). Results All tumors were located in supratentorium, including 13 cases in the temporal or parietal lobe, 1 cases in the occipital lobe, and 1 cases in the thalamus. Tumors appeared as round or ellipse in shape. The lesions ranged in size from 3 to 9 cm (mean 4.6 cm). CT scan showed mingled hypo and isodensity in 7, hypodense in 4 with CT value from 18 to 22 HU, hyperdense in 4 with the CT value from 48 to 55 HU. Enhancement appeared as an irregular pattern. The margin was clear. Most intracranial gliosarcoma showed perifocal edema. MRI were examined in 6 cases, whichappeared as low signal on T 1WI, and as high signal on T 2WI. There was irregular enhancement of the mass. Conclusion It was difficult to make the correct preoperative diagnosis for gliosarcoma, but it was possible to make a tendency diagnosis using CT or MR combined with the history. Contrast enhanced CT and MRI should be useful in differentiating gliosarcoma from glioblastoma.

In this article, we observed the serum level of Prolactin and the intensity of renal insufficiency in 40 patients with renal failure and compared their relationship. Our study showed that the serum level of Prolactin in patients with renal failure was much higher than that of normal controls (P

Phospholipase A_2 (PLA_2) inhibitor quinacrine was used to explore protective effect on multiple organ dysfunction (MOD) caused by intestinal ischemia-reperfusion (I/R) in rats. Gut I/R caused the increase of gut PLA_2 activity and induced endotoxemia and bacteriemia. Pretreatment with intravenous quinacrine 10mg?kg~(-1) attenuated bacteria and endotoxin translocation,markedly lowered the levels of thromboxane A_2 and prostacyclin I_2 in blood,and provided protection from the development of vital organs dysfunction. As a result,the survival rate in pretreatment group increased by 25%. The results demonstrate that gut I/R promotes gut barrier failure,then contributes to the development of MOD by allowing bacteria or endotoxin reaching the circulation. PLA_2 and PLA_2-dependent lipid mediators play an important role in the development of gut I/R injury and MOD. Intravenous quinacrine has protection against MOD resulting from gut I/R.