Objective:By cocculture with mesenchymal stem cells as feeder cells,we observe whether we can isolate and culture embryonic stem cells(ES cells)which maintain undifferentiated and multipotential. Methods :tissues and cell types were used in basical research to cure disease. The blastocysts of 3.5 d from Kunming species mice were collected and cultured on mesenchymal stem cells layers. The embryonic stem cell colonies were preliminarily identified by light microscope AKP staining and 0ct-4 staining in morphology and biological character. Results:The embryonic stem cell colonies by cocculture with mesenchymal stem cells as feeder cells were identified. The results showed that: 1. all the ES cells had its typical morphological characteristics such as nestlike colonies,round or elliptic,with clear edge and smooth surface,strong refraction ,significant swelling growth,compactness of cell aggragation and indistinction between two edges of neighbour cells within a colony,relatively bigger nuclear compared with little cytoplasm,i.e.,high ratio of the nuclear- cytoplasm; 2.after the AKP staining,all of the ES cells showed strong AKP activity as same as the ES cells of F0,and on the contrary,the embryonic fibroblast cells had no expression of AKP showing the lost of staining;3. after the Oct-4 staining,all of the ES cells showed strong Occt-4 activity. Conclusion:By cocculture with mesenchymal stem cells as cells,we can isolate and culture embryonic stem cells (ES cells) which maintain undifferentiated and multipotential.

Objective To observe the effect of two kinds of antibiotic therapeutic combined with humanistic care for neonates with infectious pneumonia and for intestinal micro ecology.Methods 210 neonates with infectious pneumonia and without complications from March 2011 to July 2013 were randomly divided into the cephalosporins group(n =50),the penicillins group(n =48)and the combination group(n =52),60 healthy neonates were selected as control.The cure rate,cure time in these groups were compared and the intestinal micro flora were calculated after 5 days with antibiotic treatment to evaluate the antibiotic effect to neonatal intestinal micro ecology.Results There was no significant difference between the cure rate of the cephalosporins group (76.00%),penicillins group (70.83%)and combination group(71.15%)(χ2 =2.517,P >0.05).The bifidobacteria quantity in the drug groups was significantly lower than that in the control group(t =4.600,3.634,4.275,all P <0.01),while the quantity of enteric bacilli(t =4.600,3.634,4.275,all P <0.01),enterococci(t =5.295,4.873,5.041,all P <0.01)and pep-tostreptococcus (t =3.056,2.900,3.113,all P <0.01 )in the drug groups were higher than those in the control group,but the quantity of bacteroidosis had no significant difference among the three groups (t =0.112,1.096, 1.943,all P >0.05).Conclusion Cephalosporin antibiotics and combination is not superior to penicillin,but in-clined to combined medication;antibiotics will destroy the neonatal intestinal micro ecological imbalance,endanger the health of newborn.

Objective To identify the relationship between the clinical characteristics and prognostic factors in sequentially occurring primary cancers of the breast and ovarian.Methods Clinical and pathological data of 44 cases of successive primary cancers of the breast and ovarian in each single patient diagnosed in Tianjin Medical University Cancer Institute and Hospital from 2000 to 2013 were retrospectively analyzed.Results The median overall survival of all patients with double primary cancers of breast and ovarian was 264 months.3-year overall survival rate was 95.3％ and 5-year overall survival rate was 83.0％ respectively.32 patients were first diagnosed with breast cancer and 12 patients firstly suffered from ovarian cancer.The interval between breast cancer and ovarian cancer was 1-240 months (median 60 months).The staging of ovarian cancer (P =0.005) is the main prognostic factor.In patients firstly diagnosed breast cancer,age (P =0.025) and chemotherapy(P =0.010) were mainly associated with the interval between breast cancer and ovarian cancer.Age when first cancer diagnosed,tumor size of the breast and staging of ovarian cancer were mainly prognostic factors.Age when breast cancer diagnosed and chemotherapy were factors impacting the interval of breast cancer and ovarian cancer.Conclusions For patients with sequentially diagnosed primary cancers of the breast and ovarian cancer,late stage ovarian cancer predicts a poor prognosis.The interval in patients aged 50 and below suffering from secondary cancer is long.Proper chemotherapy for breast cancer seems to delay the occuring of ovarian cancer.

Objective To study the influence of dihydromyricetin on the oxidation and non-enzymatic glycosylation in impaired glucose tolerance rats.Methods Animal model of impaired glucose tolerance rat was built by intragastrical(i.g.) injection of D-galactose generally.After 8 weeks intragastrical(i.g.) injection of dihydromyricetin(experimental group) and metformin (positive control),the levels of fasting blood glucose and tow-hour postprandial blood glucose,the insulin level and its resistance index,the levels of glycosylated hemoglobin (GHb) and advanced glycation end-products(AGEs) were detected,and the activities of superoxide dismutase(SOD) and glutathione peroxidase (GSH-Px),the content of malondiadehyde(MDA) were determined respectively.Results Compared with model group,dihydromyricetin decreased the level of two-hour postprandial blood glucose (P0.05).Conclusion Dihydromyricetin can significantly improve the state of impaired glucose tolerance rats; the mechanism may be related to its effect of improving the antioxidant activity of GSH-Px,inhibiting non-enzymatic glycation,lowering body oxidative stress and insulin resistance and promoting the use of blood glucose in peripheral organizations.

Since the first paper reported the finding of genetic variation contributing to human age-related macular de?generation by genome-wide association study (GWAS) in 2005, large number of human complex diseases associated genetic variants have been identified through GWAS method. However, the biological function of these genetic variants is not very clear. The present paper reviewed the methods of fine-mapping and the progress of the functional studies for these suscepti?bility variants. In the post GWAS Era, the study of genetic mechanisms can help us to understand the disease pathogenesis.

Objective To investigate the effects of different doses of rosuvastatin on expression of liver X receptor(LXR) and caveolin-1 in cultured human monocyte-macrophage cells which induced by oxidized low density lipoprotein (ox-LDL).Methods The human monocyte-macrophage cells were divided into six groups:control group,ox-LDL group,different doses of rosuvastatin group (0.01 μmol/L,0.1 μmol/L,1 μmol/L,5 μmol/L).The expression of LXR mRNA and caveolin-1 mRNA were assayed by RT-PCR.Results LXR mRNA expression induced by ox-LDL in the control group and ox-LDL group were 1.00 ± 0.02,0.26 ± 0.02,and the difference was significant (t =56.39,P ＜ 0.001).Meanwhile,caveolin-1 mRNA expression in ox-LDL is (0.27 ± 0.01) fold than that in control (t =31.27,P ＜ 0.001).Meanwhile,There were significant differences among ox-LDL group and the different doses of rosuvastatin group in terms of LXR mRNA and caveolin-1 mRNA expressions (F =72.154,66.007,P ＜ 0.001).Along with the increase the doses of rosuvastatin,there was an increased trends of LXR mRNA and caveolin-1 mRNA expressions (P ＜ 0.05).Conclusion Rosuvastatin and upregulated the LXR mRNA and caveolin-1 mRNA expressions in a dose dependent manner.

Objective To investigate the effects,mechanisms,and the optimum doses of Rosuvastatin and Losartan on expression of caveolin-1 in cultured human monocyte-macrophage cells which were induced by oxidized low density lipoprotein(ox-LDL).Methods Human-monocyte cells were separated and changed into the human monocyte-macrophage cells.The model of amerosclerosis was set up.These cells were incubated in different doses of Rosuvastatin(0.1,1.0,5.0 μmol/L) and Losartan (10,50,100 μmol/L),and then cultured in combination of two drags (5.0 μmol/L + 100 μmol/L).Expression of caveolin-1 mRNA was determined with real-time fluorescent quantitative polymerase chain reaction (RT-PCR).Results In ox-LDL group,caveolin-1 mRNA was decreased sharply relative to control group [(0.2533 ±0.00973) vs (0.9410 ±0.03677)] in a concentration-dependent manner (P ＜0.01).Compared to ox-LDL group,expressions of Caveolin-1 mRNA were increased gradually in different doses of Rosuvastatin alone and Losartan alone group [(0.5198 ± 0.04840),(0.6183 ± 0.06740),(0.7257 ± 0.03052) vs (0.2533 ± 0.00973) ; (0.3350 ± 0.04177),(0.4428 ± 0.03804),(0.6049 ± 0.02627) vs (0.2533 ± 0.00973)] in a concentration-dependent manner (P ＜ 0.01) ; the summit expressions of caveolin-1 mRNA were emerged in using Rosuvastatin and Losartan together (F =59.119,P ＜ 0.01).Conclusions Rosuvastatin and Losartan may be responsible for the expression of caveolin-1 in human monocyte-macrophage cells that were induced by ox-LDL.The expressions were up-regulated with dose dependent manner of these drugs,and got the crest stage when using optimum doses of Rosuvastatin and Losartan together.

Objective To evaluate the function status of motor nerve in patients with carpal tunnel syndrome ( CTS) sensory neuropathy by single-fiber conduction studies ( SF-CS).Methods Forty patients with CTS were divided into two groups according to the nerve conduction abnormality of median nerve .The sensory conduction abnormalities ( SCA) group included 20 patients with abnormal sensory conduction , while the sensory motor conduction abnormalities ( SMCA) group included 20 patients with abnormal motor sensory conduction.The Keypoint.net electromyogram device was used to detect the wrist to the abductor pollicis brevis latency , using the saddle shaped stimulating electrodes for stimulating and the single fiber electromyography electrode for recording.The control group included 20 healthy people.Results The latency of single-fiber conduction of median nerve in SCA group , SMCA group and control group was (3.92 ±0.28) ms, (4.71 ±0.49) ms and (3.41 ±0.31) ms (F=63.829, P=0.000), respectively.The area under the receiver operating characteristic curve was 0.920 for the latency of single-fiber conduction of median nerve , and its specificity was 85%in cut off point.Conclusions There is clinical motor nerve damage in patients with CTS sensory neuropathy.The SF-CS may become an effective means for early evaluation of motor nerve function state.

Objective:To explore the e ect of QingreJiedu herbs in combination with Jiegeng decoction on expression of NF-?B p65 in ALI induced by LPS.Methods:Male Sprague-Dawley rats were randomly divided into normal group,model control group,QingreJiedu group,and the group of QingreJiedu herbs in combination with Jiegeng decoction.The model of ALI was made by injected with a single dose of LPS through vena femorali,the pathologic morphology,MPO activity and the expression of NF-?B p65 was observed.Result:Compared with normal group,infiltration.of PMNs,capillary congestion and swelling were found in model control group,and the content of MPO and expression of NF-?B p65 in the nuclear were significantly increased(P

Objective To explore the effects of biomimetic electrical stimulation on inducing differentiation of rat bone marrow mesenchymal stem cells(MSCs) into cardiomyocyte-like cells in isolated myocardium.Methods MSCs and cardiac myocytes from SD rats were isolated and cultured by modified method.MSCs were then co-cultured with cardiac myocytes(MSCs/ cardiomyocyte co-culture system).The co-culture system was divided into stimulated group and non-stimulated group.MSCs cells in stimulated group were given supra-threshold square biphasic pulses stimulation(5ms duration,0.5Hz,10V) lasting for 2h each day,while in non-stimulated group were routinely cultured.Each group was divided into three subgroups according to the incubation time of 3,5 and 7 days.The characteristic morphology of MSCs in every subgroup was observed under light microscope.The expression of cTnT in MSCs was detected by immunofluorescence.The rate of cTnT positive expression was also compared between two groups.Results The growth and frequency of myocardial wave of MSCs cells in stimulated group were better or higher than those in non-stimulated group.No expression of cTnT was detected in non-stimulated group at the 5th day of the experiment,while a few MSCs cells in the stimulated group expressed cTnT at the same time point.The cTnT expression was detected in the MSCs cells in the both groups on the 7th day.The rate of cTnT positive expression in stimulated group(20.98%?5.55%) was significantly higher than that in non-stimulated group(13.20%?3.98%,P