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Background@#The emergence of the severe acute respiratory syndrome coronavirus 2 omicron variant has been triggering the new wave of coronavirus disease 2019 (COVID-19) globally. However, the risk factors and outcomes for radiological abnormalities in the early convalescent stage (1 month after diagnosis) of omicron infected patients are still unknown. @*Methods@#Patients were retrospectively enrolled if they were admitted to the hospital due to COVID-19. The chest computed tomography (CT) images and clinical data obtained at baseline (at the time of the first CT image that showed abnormalities after diagnosis) and 1 month after diagnosis were longitudinally analyzed. Uni-/multi-variable logistic regression tests were performed to explore independent risk factors for radiological abnormalities at baseline and residual pulmonary abnormalities after 1 month. @*Results@#We assessed 316 COVID-19 patients, including 47% with radiological abnormalities at baseline and 23% with residual pulmonary abnormalities at 1-month follow-up. In a multivariate regression analysis, age ≥ 50 years, body mass index ≥ 23.87, days after vaccination ≥ 81 days, lymphocyte count ≤ 1.21 × 10 -9 /L, interleukin-6 (IL-6) ≥ 10.05 pg/mL and IgG ≤ 14.140 S/CO were independent risk factors for CT abnormalities at baseline. The age ≥ 47 years, presence of interlobular septal thickening and IL-6 ≥ 5.85 pg/mL were the independent risk factors for residual pulmonary abnormalities at 1-month follow-up. For residual abnormalities group, the patients with less consolidations and more parenchymal bands at baseline could progress on CT score after 1 month. There were no significant changes in the number of involved lung lobes and total CT score during the early convalescent stage. @*Conclusion@#The higher IL-6 level was a common independent risk factor for CT abnormalities at baseline and residual pulmonary abnormalities at 1-month follow-up. There were no obvious radiographic changes during the early convalescent stage in patients with residual pulmonary abnormalities.
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Objective:To evaluate the correlation between brain-derived neurotrophic factor (BDNF) and inflammatory markers in rheumatoid arthritis (RA) patients with depressive symptoms.Methods:This study was a cross-sectional study. RA patients' medical history were recorded and disease activity was evaluated. Serum BDNF, interleukin (IL)-6, tumor necrosis factor (TNF)-α were tested and clinical inflammatory indicators such as C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), fibrinogen (FIB), serum amyloid A (SAA) were recorded. RA patients were instructed to fill in the patient health Questionnaire-9 (PHQ-9) scale by themselves. Patients with a score greater than or equal to 5 were included in the RA with depressive symptoms group, and patients with a score of 4 or less were included in the RA without depressive symptoms group. The changes in BDNF and inflammatory indexes were compared between the two groups. Correlation analysis of PHQ-9, BDNF, inflammatory markers and DAS28 was performed. Logistic regression analysis was performed to find the risk factors of depression in RA.Results:A total of 140 RA patients were enrolled in this study, and 66 patients (47.1%) with a PHQ-9 score greater than or equal to 5 were included in the RA with depressive symptoms group. Compared with the RA without depressive symptoms group, RA patients with high disease activity, single and living alone, poor economic self-awareness and unemployed were more likely to have depressive symptoms. The serum level of BDNF[(2 276±333) pg/ml vs (1 367±431) pg/ml, t=13.91, P<0.001], IL-6[(39±28) pg/ml vs (27±8) pg/ml, t=3.66, P<0.001], TNF-α[(9.0±7.2) pg/ml vs (6.6±3.9)pg/ml, t=2.43, P=0.035], CRP[(25±13) mg/L vs (17±11) mg/L, t=3.94, P<0.001], ESR[(48±18) mm/1 h vs (34±21) mm/1 h, t=4.14, P=0.024], Fib[(3.8±1.1) g/L vs (3.0±0.5) g/L, t=5.92, P=0.023], SAA[(64±39) mg/L vs (37±19) mg/L, t=5.32, P<0.001] in RA with depressive symptoms group were significantly higher than those in RA without depressive symptoms group. Serum BDNF was significantly positively correlated with PHQ-9 score ( r=0.66, P<0.001), IL-6( r=0.20, P=0.019), TNF-α ( r=0.14, P=0.090), CRP ( r=0.32, P<0.001), ESR ( r=0.20, P= 0.001), Fib ( r=0.28, P=0.001), SAA( r=0.28, P=0.001) and DAS28 ( r=0.37, P<0.001) . BDNF [ OR (95% CI) =1.578(1.257, 2.354), P=0.001], IL-6[ OR (95% CI) =1.073(1.012, 1.075), P=0.006], CRP[ OR(95% CI)=1.085(1.045, 1.178), P=0.001], SAA[ OR(95% CI)=1.125(1.004, 1.198), P=0.018] and unemployment were risk factors for depressive symptoms in RA. Conclusion:Serum BDNF is positively correlated with PHQ-9 scores, inflammatory markers and disease activity in RA patients. BDNF, IL-6, CRP, SAA and unemployment are risk factors for depressive symptoms in RA. Effective treatment of RA can reduce the occurrence of depression symptoms.
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Objective:To analyze the causes of death of elderly inpatients,so as to provide direction for the precaution of geriatrics.Methods:The first page data of hospitalized medical records of elderly inpatients in medical department in the Second Hospital of Dalian Medical University from 2102 to 2019 were extracted, and the causes of death were analyzed. The statistical analysis was performed by Excel and SPSS 21.0 statistical software.Results:The number of deaths of elderly inpatients in medical department from 2012 to 2019 was 5 249. The proportion of deaths in ICU was 34.43%(1 807/5 249), in oncology department was19.03%(999/5 249), and in cardiovascular department was12.08%(634/5 249). The average age was (78.52 ± 7.82) years. Besides, the age of men was younger than women: (78.18 ± 8.00) years vs. (79.02 ± 7.52) years, and the differences was statistically significant ( P<0.01). Men(59.1%, 3 099/5 249) were more than women (40.96%, 2 150/5 249). The largest number of deaths was in the age of 75-84 years (42.56%, 2 234/5 249). The number of cases with combined above five diseases was 4 552(86.72%, 4 552/5 249). The top three causes of deaths of elderly inpatients in medical department from 2012 to 2019 were cardiocerebrovascular diseases (27.21%, 1 428/5 249), malignant tumor (25.74%, 1351/5 249) and respiratory system diseases (22.10%, 1160/5 249). From 2012 to 2015, the top three causes of deaths were malignant tumor, cardiocerebrovascular diseases and respiratorysystem diseases. From 2016 to 2019,the top three causes of deaths were cardiocerebrovascular diseases, malignant tumor, and respiratory system diseases. The most common cause in cardiocerebrovascular diseases of death was coronary heart disease (51.47%,735/1 428), cerebrovascular disease (43.70%,624/1 428),and hypertension(4.34%, 62/1 428). Among the patients with malignant tumor death, first cause of death waslung malignant tumor (37.53%, 507/1 351), the others in turn were gastric carcinoma (11.10%,150/1 351) and intestinal cancer (11.10%,150/1 351). Among the patients with respiratory system diseases, first cause of death was pulmonary infection (69.66%,808/1 160), the others in turn were chronic obstructive pulmonary disease (15.43%, 179/1 160) and interstitial lung disease (5.09%, 59/1 160). Conclusions:The average age of elderly inpatients in medical department is (78.52 ± 7.82) years. The death age of male is less than that of female, and the number is slightly more than that of female. The top three causes of deaths of elderly inpatients in medical department are cardiocerebrovascular diseases, malignant tumor, respiratorysystem diseases. From 2012 to 2015 the first cause of deaths is malignant tumor. From 2016 to 2019, the cardiocerebrovascular diseases rise to the first.
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Objective:To evaluate the clinical characteristics and identify potential factors of the early-stage hip involvement in patients with ankylosing spondylitis (AS) based on the magnetic resonance imaging (MRI).Methods:A retrospective group control study was carried out in 570 AS patients who were consecutively admitted to our hospital from 2014 to 2018. Patients with hip pain or hip function limitation but lacking definitive evidence of hip involvement on radiography were underwent hip MRI. Patients were divided into three groups: no hip involvement, early-stage hip involvement (hip involvement detected by MRI but with negative radiographs) and advanced-stage hip involvement (Bath Ankylosing Spondylitis Radiology Index-hip score ≥2). The study factors included demographic, laboratory, clinical and radiographic data. Simple and multiple ordinal logistic regression analysis were used to identify factors associated with the early-stage hip involvement and advanced-stage hip involvement.Results:A total of 236 patients (41.4%) presented with hip involvement, in which 146 cases (25.6%) were diagnosed with early-stage hip involvement, while 90 cases (15.8%) were diagnosed with advanced-stage hip involvement. Multivariate logistic regression analysis demonstrated that older age at onset [ OR=0.80, 95% CI (0.72, 0.90), P<0.01], more active inflammation in the sacroiliac joints [ OR=1.13, 95% CI(1.07, 1.18), P<0.01] and worse BASMI score [ OR=3.06, 95% CI(2.14, 4.13), P<0.01] were associated with the occurrence of early-stage hip involvement. Conclusion:MRI is superior to radiography in detecting early-stage hip involvement. MRI is more suitable for hip involvement assessment in AS patients with suspected symptoms or risk factors of hip involvement.
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Objective To observe the effect of Tingli-Dazao-Xiefei decoction combined with chemotherapy in the treatment of lung cancer with pleural effusion. Methods A total of 90 patients who met the inclusion criteria from January to June 2017, were randomly divided into treatment group and control group. All patients were treated with intrapleural administration of cisplatin chemotherapy once a week, while patients in treatment group received additional treatment of taking Tingli-Dazao-Xiefei decoction daily.Changes of clinical efficacy, traditional Chinese medical (TCM) symptom score and Karnofsky score were observed 4 weeks later. Results After 4 weeks of treatment, there was no difference in the total effective rate between the two groups (χ2=1.600, P=0.659). The total effective rate was 71.1% (32/45) in treatment group, including 11 cases of complete remission and 20 cases of partial remission, while it was 64.4% (29/45) in control group including 10 of complete remission and 19 of partial remission. After treatment, the TCM symptom scores in the treatment group(20.81 ± 1.92 vs.8.93 ± 1.27;t=34.619,P<0.001)and control group(20.28 ± 1.36 vs.13.22 ± 1.63; t=22.310, P<0.001) were significantly lower than those before the treatment. After treatment, the TCM symptom scores(8.93 ± 1.27 vs. 13.22 ± 1.63,t=13.927)in the treatment group were significantly lower than that in the control group(P<0.001).The Karnofsky score(95.6% vs. 80.0%,χ2=3.728)in the treatment group was significantly higher than that in the control group (P<0.05). There was no difference in the incidence of adverse reactions. Conclusions The theray of Tingli-Dazao-Xiefei decoction combined with chemotherapy could achieved the similar clinical effect of cisplatin chemotherapy treatment, while it showed the advantage of improving TCM syndrome score and the quality of life.
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Objective To investigate the risk factors of disease progression and adverse pregnancy outcome in patients with systemic lupus erythematosus (SLE) during pregnancy.Methods Clinical data of 118 pregnant women with SLE admitted from June 2004 to October 2015 were retrospectively analyzed.The patients were divided into selective pregnant group (group A,n =72 cases) and non-selective pregnant group (group B,n =46) according to the disease activity of SLE.The disease progression and pregnancy outcomes were compared between two groups.Results The various system damages occurred in group B,including hematological damage in 16 cases,kidney damage in 19 cases,erythra in 10 cases,arthritis in 10 and serositis in 12 cases;while the corresponding cases in group A were 10,14,6,4 and 4(x2 =7.133,P=0.008;x2 =6.658,P =0.010;x2 =4.304,P =0.038;x2 =7.030,P =0.008;x2 =10.095,P =0.001).SLE exacerbation occurred in 28 cases (74％) of group B and 12 cases (17％) of group A (x2 =34.944,P =0.000).The logistic regression analysis showed that hypocomplementemia,proteinuria,SLEDAI score before pregnancy and positive anti-dsDNA antibody were the risk factors of SLE disease exacerbation during pregnancy.The maternal complications occurred in group B,including pregnancyinduced hypertension in 7 cases,preeclampsia in 10 cases and infections in 11 cases/times;while the corresponding cases (case/time) in group A were 2,6 and 4 (x2 =4.526,P =0.033;x2 =4.304,P =0.038;x2 =8.525,P =0.004).There were 14 cases of therapeutic induced labor,7 case of stillbirth and 27 cases of total fetal loss in group B,while the corresponding cases in group A were 2,0 and 4 (x2=18.317,P =0.000;x2 =9.080,P =0.003;x2 =40.920,P =0.0300).The logistic regression analysis showed that positive anticardiolipin antibody,proteinuria,SLEDAI score before pregnancy and renal dysfunction during pregnancy were risk factors of fetal loss.There were 87 cases of successful delivery (73.7％),the successful delivery rates were 41.3％ (19/46) in group B and 94.4％ (68/72) from group A,respectively.The infant complications occurred in group B,including premature birth in 15 cases,low birth weight in 13 cases,neonatal jaundice in 5 cases and mild asphyxia in 5 cases;while the corresponding cases in group A were 24,18,4 and 2 (x2 =11.442,P =0.001;x2 =11.395,P =0.001;x2 =4.664,P =0.031;x2 =8.035,P =0.005).Conclusion SLE patients whose disease conditions are not well controlled would lead to higher percentage of disease deterioration during pregnancy and worse pregnancy outcomes.
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Objective To find out whether NOTCH receptors can serve as direct downstream targets of transforming growth factor β(TGF-β)/Smad4 signaling in endothelial cells.Methods Real-time PCR and Western blotting were performed to verify whether the expression of notch1 and notch4 was regulated by TGF-β pathway.Luciferase reporter assay was employed to investigate how the promoter of notch1 and notch4 was regulated by TGF-β.Then, ChIP assay was used confirm whether the promoter of notch1 and notch4 physically interacted with SMAD protein.Results TGF-β1 and bone morphogenetic protein 4 (BMP4) treatment increased the expression of both notch1 and notch4 at the transcriptional level.In addition, SMAD4 was physically associated with the SMAD binding sites on the notch4 promoter, which was largely enhanced under the treatment of TGF-β1 and BMP4.Importantly, TGF-β1 and BMP4 failed to transactivate notch4 in the absence of endogenous SMAD4 or the SMAD binding regions on the notch4 promoter.Conclusion The expression of NOTCH receptor can be directly up-regulated by SMAD4-mediated TGF-β/BMP signaling in cerebrovascular endothelial cells.
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Objective To investigate the function of CXCR4 and CXCR7 receptors expressed in bone marrow mesenchymal stem cells (BMSCs) membrane surface in process of cell migration,in order to provide a theoretical basis for bone trauma repair.Methods C57BL/6 mice were selected to collect BMSCs of second passage after using the adherence culture method.Expressions of CXCR4 and CXCR7 receptors on BMSCs membrane surface were detected using immunofluorescent staining and flow cytometry.In CXCL12-induced chemotaxis of BMSCs,the assay was divided into control group (cells were seeded in serum-free medium),CXCL12 group and (cells were seeded in serum-free medium containing CXCL12),and CXCR4-blocked group (cells were seeded in serum-free medium containing CXCL12 after the blockade of CXCR4).Migration of BMSCs was qualified and used to determine the chemotaxis role of CXCR4 and CXCR7.After the blockade of CXCR4,expression of CXCR7 was detected using the Western blot method.Results lmmunofluorescence showed overexpressions of CXCR4 and CXCR7 receptors on BMSCs membrane surface.Flow cytometry showed the positive rate of CXCR4 and CXCR7 on BMSCs membrane surface was 96.4％ and 97.3％ respectively.Cell migration assay showed amount of MBSCs migration was the highest in CXCL12 group,relative higher in CXCR4-blocked group and the lowest in control group (P ＜ 0.01).In CXCR4-blocked group,expression of CXCR7 increased.Conclusion CXCR4 and CXCR7 Receptors are expressed in BMSCs membrane surface,but CXCR4 play the major role in CXCL12-induced BMSCs migration to traumatic bone wounds.
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Objective To study the risk factors for dose wearing off (WO)in PD patients .Methods One hundred and thirty-three PD patients were recruited in this study according to the UKPDBB criteria .Their dose WO was diagnosed according to the Wo questionnaire 9 (WOQ-9) .The pa-tients were divided into WOQ-9 (+ ) group (n=111) and WOQ-9 (-) group (n=22) .The pa-tients in WOQ-9 (+) group were further divided int WO (+ ) group (n=59) and WO (-) group (n=52) .The difference in their clinical and therapeutic parameters was compared .Results The dose WO was observed in 83 .5% of the 133 PD patients ,53 .2% of which accorded with the dose WO clinical definition .T he disease onset age ,disease course ,maximal levodopa daily dose and ac-cumulated levodopa dose differed greatly in WOQ-9 (+ ) group and WOQ-9 (-) group (P<005) . The disease course ,H-Y stage ,UPDRS score ,tetany score ,maximal levodopa daily dose ,levodopa dose per body weight and accumulated drug use time differed greatly in WOQ-9 (+ ) group and WOQ-9 (-) group (P< 0 .01) .The major risk of dose WO was the levodopa dose per body weight (OR=1.364 ,P<0 .05) .Conclusion Dose WO is related with the progress of disease and the use of levodopa .Levodopa dose per body weight is an independent risk factor for dose WO .
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Objective To explore the risk factors of dyskinesias in Parkinson' s disease (PD) patients.Methods Patients with PD who had taken levodopa at least 1 month were recruited according to the United Kingdom Brain Bank (UKBB) Criteria..All patients were divided into dyskinesias group and non-dyskinesias group according to the clinical definition of dyskinesia.The parameters including gender,age,age onset,duration,body weight,UPDRS scale score,and treatment parameter,such as the maximum daily dose of levodopa,the cumulative time of medication of levodopa,levodopa dose per weight of patients with dyskinesias were recorded.Multivariate logistic regression analysis was used to analyze risk factors of dyskinesias.The patients were divided into dyskinesias and no dyskinesias groups based on presence of motor complications.Results The incidence rate of dyskinesias was 7.8％ (11/142) in all 142 patients.Of which,9 cases were with peak-dose dyskinesia,and 2 cases with both wearing-off and dyskinesia.There were statistically significant difference between dyskinesias group and non-dyskinesias group in terms of sex,weight,the maximum daily dose of levodopa and levodopa dose per weight(P ＜ 0.05).The same results appeared between wearing-off group and wearing-off with dyskinesia group (P ＜ 0.05).Multivariate Logistic regression analysis showed that total dose of levodopa(OR =1.846,95％ CI:1.234-2.762,P =0.003) and levodopa dose per weight were independentriskfactors(OR=0.991,95％CI:0.984-0.999,P=0.033).Conclusion The risk factors of dyskinesias in Parkinson's disease is closely linked to total dose of levodopa and levodopa dose per weight.
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Objective To observe the histopathologic injury of small intestine and intestinal permeability in chronic renal failure (CRF) rats. Methods Twenty male Sprague-Dawley rats were randomly assigned to CRF group (n=10) and control group (n=10). 5/6 nephrectomy was used to establish CRF rats, while sham operation for control. Blood biochemistry was regularly monitored until CRF model was successfully established. The model rats were fed with lactulose (L) and mannitol (M) through intragastric administration. Urine was collected after 6 hours, and the concentration of lactulose and mannitol in urine was measured using high pressure liquid chromatograph with refractive index detector (HPLC-RID), and the ratio of urinary excretion of L/M was calculated to evaluate intestinal permeability. Small intestinal mucosa were stained by hematoxylin-eosin (HE) and observed with light microscope (villus height, thickness of muscle layer and villus count), histological damage score was used to evaluate intestinal injury. Results The L/M ratio of CRF group was higher than that of control group (1.75±0.11 vs 1.20±0.06, P<0.01). The small intestinal mucosal villus height and thickness of muscle layer in CRF group were higher (P<0.01), and the number of villi was lower compared to control group (P<0.01). The score of histopathologic intestine damage of CRF group was higher than that of control group (1.00±0.71 vs 0, P<0.01). Conclusion The intestinal permeability of CRF rats is increased with varying degrees of intestinal damage.
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AIM:Studies on totipotential stem cells using primordial germ cells (PGCs) have the same application prospect as embryonic stem cells, especially for the animals difficult to isolate embryonic stem cells from blastula. In vitro culture condition of embryo germ cells is the key to control this. This study designs a method to effectively amplify PGCs of mice in vitro and establishes an effective culture method of PGCs. METHODS: Experiments were carried out in the Department of Histology and Embryology of Chongqing Medical University and Key Laboratory of Biochemistry and Molecular Pharmacology from October 2006 to August 2007. ①Clean Kunming pregnant mice (embryos 0.5 dpc) were provided by Animal Experimental Center of Chongqing Medical University. Experimental procedures met the animal ethical standards. ②Allantois and surrounding tissues of 8.5 day post coitum (dpc) embryos, the hindhut and surrounding tissues of 9.5 dpc and 10.5 dpc embryos, the gonadal ridges and surrounding tissues of 11.5 dpc and 12.5 dpc embryos were collected and digested with 0.25%pancreatin-0.02%EDTA, then the cells were cultured in the plastic petridishes which are pretreated with 0.1% gelatine. The formation ratio of primary and the first passaged Embryonic germ (EG) clones were compared among those different time points. The SSEA-1 positive ratio of isolated cells was compared between the two days by immunohistochemical method. The assays of clone numbers counting and MTT were used to compare the different proliferation ability of PGCs from the 11.5 dpc and 12.5 dpc embryos. The effects of expanding PGCs from 11.5 dpc and 12.5 dpc embryos by those two ways above were compared. The EG clones were analyzed by the expression of alkaline phosphatase and the immunologic marker SSEA-1,the undifferentiated marker Nanog and the differentiation ability in vitro were also tested. RESULTS:①The formation ratio of primary and the first passaged EG clone from 11.5 and 12.5 dpc embryos was higher than that from 8.5 to 10.5 dpc embryos and efficiency of expansion was increased (P 0.05). On the third day, the number of the primary EG clones from 11.5 dpc embryos was higher than that from 12.5 dpc embryos (P 0.05), but the two groups had different characteristics. ④The EG clones have representative morphology with high level of alkaline phosphatase activity and SSEA-1, Nanog expression, which can differentiate into the cystic embryoid body. CONCLUSION: 11.5 dpc and 12.5 dpc mouse embryos, especially 11.5 dpc embryos are the optional time points to expand PGCs efficiently. Co-culturing with the tissue and isolating the gonadal ridges both of these two ways are practicable.
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Physiology is the main course in the education of preclinical medicine.And the functional experiment education is one of the important ingredients in it.The teaching style should be open,the students capability of independent thoughts should be raised and at the same time their precise scientism and cooperation spirit should be cultivated following the basic outline of teaching.
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Objective:To clone LIF gene for the construction of mammalian expression vector PCDNA3.1(+)-LIF.Methods:LIF gene was cloned from pregnant uterine decidua tissues by using RT-PCR method,the gene was ligated with T-vector,then LIF gene was excised from T-LIF plasmid and inserted into PCDNA3.1(+) to construct eukaryotic expression vector consisting of LIF gene.Results:LIF gene sequence and vector were verified correctly by enzyme digestion,sequence analysis.Conclusion:The successful clone of LIF gene and the construction of its mammalian expression vector has been obtained.The results have provided basis for the further studies on LIF.
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Objective To study the spatiotemporal distributions of epithelial proliferation and apoptosis,and the correlation of morphogenesis with them in developing stomach.Methods Serial sections of fetal mice aged 11-15.5 days were made.The morphogenesis of stomach was observed under light microscope.The density of mitotic cells(DMC) and apoptotic bodies(DAB) of epithelium in the developing cardia,pylorus,greater curvature,lesser curvature were measured with stereological method.Results The morphological development of stomach took place on the 11~(th) day of mouse embryo development.The fundus,greater curvature and lesser curvature were observed on the 12~(th) day.The peak value of DMC all appeared before the peak value of DAB.In the epithelium of cardia,no obvious peak value of DMC was observed and the peak value of DAB was on the 11.5~(th),12.5~(th),13.5~(th) day.In the epithelium of pylorus,the peak value of DMC was on the 12.5~(th) day,the peak value of DAB was on the 14~(th) day.In the epithelium of greater curvature,the peak value of DMC was on the 11.5~(th) and 12~(th) day,the DAB value was low.In the epithelium of lesser curvature,the peak value of DMC was on the 11.5~(th) day,the peak value of DAB was on the 12.5~(th) day.Conclusion There was very close spatiotemporal relationship between the cell proliferation,cell apoptosis and the morphological development of stomach.
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Objective To investigate the isolation, purification and culture methods of human embryonic germ cells (EGCs) on feeder layer cells of human embryonic fibroblasts. Methods Primordial germ cells(PGCs) from the genital ridges of 6-11 weeks(post fertilization) human embryos were cultrued on feeder layer cells of human embryonic fibroblasts(HEFs) which were isolated from the 3-4 month embryos and routinely cultured for over 25 passages. The medium composed of growth factors and differentiation inhibitory factors. The cultures were analyzed with the expression of alkaline phosphatase, immunologic markers (SSEA-1,SSEA-4) and the transcription factor Oct-4 that have been used to routinely to characterize EGCS. Results A large-scale EG cells were obtained and meintained on feeder layers for over 8 passages. The cell surface marker showed that the EGCs possess high levels of AP activity. EGCs colonies stained strongly for SSEA-4,SSEA-1 and they expressed the transcription factor Oct-4.Conclusion EGCs have potential to maintain long term proliferation and undifferentiation state on human embryonic fibroblasts in vitro.;
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Objective To study the temporal and spatial expression of Nanog in developing primordial germ cells(PGCs) of mouse.Methods Immunofluorescence technique was taken to qualitatively analyze the expression of Nanog in PGCs of 7.5days(post-coitus days)-15.5days mouse embryo.A further quantitative analysis of Nanog expression change in PGCs of 11.5days-15.5days mouse embryo was done by using SSEA-1 and Nanog double labelling immunofluorescence staining with confocal microscopy.Results The PGCs in mouse allantois,hindgut,dorsal mesentery and genital ridge were Nanog positive.Both of the highest positive ratio and the highest fluorescence intensity appeared in PGCs of 11.5days mouse embryo.For the female mice,Nanog expression drops dramatically after 12.5days,and for the male mice,a noticeable decline of Nanog expression was occurred after 13.5 postcoitus days.Conclusion\ Nanog expresses stably in the proliferating PGCs.The down-regulation of its expression may be related with the differentiation of PGCs.