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Objective:To access the independent associations between serum ferritin quintile levels and metabolic syndrome (MS) in adults of different genders.Methods:19 563 participants over the age of 18 years were recruited from "TCLSIH Cohort Study" from 2007 to 2015. Serum ferritin concentration was measured by Enzyme-linked immunoassay, while metabolic syndrome was diagnosed according to MS diagnostic criteria formulated by Chinese Diabetes Society in 2013. Multiple logistic regression analysis was used to assess the association between serum ferritin quintile levels and the prevalence of MS in males and females. Results:After adjusting the confounding factors, the overall prevalence of MS gradually increased with the increasing of serum ferritin levels, similar results were observed in males and females. Subjects were divided into 5 subgroups according to serum ferritin levels. Compared with level 1 group, logistic regression showed that the serum ferritin quintiles of males and females ranged from low to high, the OR (95% confidence interval) for metabolic syndrome were 1.142 (0.998, 1.307), 1.382 (1.210, 1.579), 1.680 (1.472, 1.917), 2.085 (1.827, 2.380), respectively ( Ptrend<0.01), and 1.147 (0.911, 1.444), 1.346 (1.075, 1.687), 1.567 (1.268, 1.941), 2.444 (1.981, 3.023), respectively ( Ptrend<0.01). Conclusion:The elevated ferritin levels were positively related to the prevalence of metabolic syndrome in adults of different genders.
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Objective@#To investigate the effects of circular RNA hsa_circ_0063772 on the proliferation, migration and invasion of oral squamous cell carcinoma (OSCC) cells.@*Methods@#Thirty-three patients with oral squamous cell carcinoma who were admitted to the Department of Oral and Maxillofacial Surgery, Peking University Shenzhen Hospital from February 2017 to December 2018 were enrolled in this study. Real-time quantative polymerase chain reaction was used to detect the expression level of circular RNA hsa_circ_0063772 in OSCC and corresponding adjacent tissues, OSCC cell lines and human keratinocytes. The expression level of hsa_circ_0063772 was overexpressed in SCC15 and CAL27 cells by using lentivirus, and the effects of this gene on proliferation, migration and invasion of OSCC cells were detected by cell counting assay, scratch assay, Transwell assay, Western blotting and nude mice tumor formation assay.@*Results@#The expression of circular RNA hsa_circ_0063772 in OSCC tissues (9.38±0.34) was lower than that in adjacent tissues (11.30±0.31) (t=5.20, P<0.001), and the expression in OSCC cells (SCC15: 0.12±0.01; SCC25: 0.18±0.02; SCC9: 0.21±0.01; CAL27: 0.13±0.01) was significantly lower than that in human keratinocytes (1.02±0.02) (tSCC15=41.91, tSCC25=29.21, tSCC9=35.16, tCAL27=41.86, P<0.001). Overexpression of hsa_circ_0063772 in SCC15 and CAL27 cells can affect tumor cell proliferation, cell counting assay showed that tumor cell proliferation ability in high expression group (SCC15: 0.76±0.01; CAL27: 0.74±0.02) were lower than empty group (SCC15∶1.22±0.04; CAL27: 0.99±0.03; tSCC15=12.58, tCAL27=6.97; P<0.05). Transwell migration experiment showed the number of migrated cell in high expression group (SCC15∶148.00±14.57; CAL27: 243.00±13.00) were less than empty group (SCC15: 580.30±42.91; CAL27: 424.70±18.66, P<0.01); Transwell invasion assay showed the number of invased cell in high expression group (SCC15: 123.70±6.98; CAL27: 326.00±17.01) were less than empty group (SCC15: 517.70±9.96; CAL27: 454.30±8.09, P<0.01). The results of tumor formation in nude mice showed that the tumor volume and mass of the overexpressed group [(306.40±16.51) mm3; (289.40±11.44) mg] were lower than that of the empty group [(582.60±32.51) mm3, t=7.58, P<0.05; (599.60±21.27) mg, t=7.58, P<0.001].@*Conclusions@#Compared with adjacent tissues, hsa_circ_0063772 is lowly expressed in oral squamous cell carcinoma. High expression of hsa_circ_0063772 can inhibit the proliferation, migration and invasion of OSCC cells.
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Objective@#To investigate the effect of long non-coding RNA highly upregulated in liver cancer (lncRNA HULC) on the biological behavior of oral squamous cell carcinoma (OSCC) cell lines.@*Methods@#A total of thirty patients with oral squamous cell carcinoma at Peking University Shenzhen Hospital from March 2017 to March 2018 were enrolled in this study. OSCC and adjacent tissues were extracted during tumor extensive resection. Quantitative real-time PCR (qPCR) was used to detect the expression levels of lncRNA HULC in OSCC and paracancerous tissues and OSCC cell lines. SCC15 and SCC25 cells were transfected with siRNA, and the effects of the gene on the biological behavior of OSCC cells were detected by cell counting assay, scratch assay, Transwell assay and Western blotting.@*Results@#The expression of lncRNA HULC in OSCC tissues (10.98±0.31, n=30) was significantly higher than that in paracancerous tissues (8.39±0.31, n=30) (t=5.93, P<0.001), the expression of lncRNA HULC in OSCC cells (SCC15: 28.58±2.74; SCC25: 16.56±0.87; SCC9: 11.18±1.32; CAL27: 13.92±0.99, n=5) was significantly higher than that in human keratinocytes (1.01±0.00, n=5) (tSCC15=10.08, tSCC25=17.96, tSCC9=7.71, tCAL27=13.09, P<0.001). Down-regulation of lncRNA HULC in SCC15 and SCC25 cells can inhibit the proliferation, migration and invasion of tumor cells and promote tumor cell apoptosis.@*Conclusions@#lncRNA HULC is highly expressed in OSCC and can enhance the proliferation, migration and invasion of cancer cells and inhibit tumor cell apoptosis.
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Objective To investigate the clinical effects of somatostatin combined with Qingyi decoction in the treatment of acute pancreatitis. Methods 120 patients with acute pancreatitis were chosen as observation subjects,and they were randomly divided into control group and observation group according to the digital table. The control group was treated with somatostatin,the observation group was treated with Qingyi decoction on the basis of control group. The clinical effects,improvements of clinical indicators and symptom,adverse reaction between the two groups were compared. Results The total effective rate of the observation group(100. 00%) was higher than that of the control group(90. 00%),the difference was statistically significant (χ2 =6. 316,P<0. 05). Before treatment, there were no statistically significant differences between the two groups in clinical indicators ( t =0. 047,0. 069, 0. 040,0. 279,0. 064,0. 101,0. 333,all P>0. 05). After treatment,the albumin of the observation group was higher than that of the control group,white blood cell count,APACHEⅡ score,C reactive protein,serum amylase,aspartate aminotransferase and lactate dehydrogenase were lower than those in the control group, the differences were statistically significant (t=14. 822,11. 654,15. 849,20. 000,10. 509,15. 213,15. 191,all P <0. 05). The blood amylase,abdominal pain relief,recovery of gastrointestinal function,urinary amylase,abdominal distension,ventilator weaning,abdominal tenderness disappeared and so on symptom improvement time of the observation group were earlier than those of control group,the hospital stay was shorter than that of the control group,the differences were statistically significant (t=10. 479,15. 855,8. 705,13. 064,8. 965,11. 783,11. 347,17. 069,7. 058,all P<0. 05). The incidence rate of adverse reactions in the control group was higher than that in the observation group(11. 67% vs. 1. 67%),the difference was statistically significant (χ2 = 4. 821, P < 0. 05 ). Conclusion Qingyi decoction combined with somatostatin can effectively improve the clinical symptom of acute pancreatitis,and with good clinical effects and safety.
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Objective To investigate the clinical effects of somatostatin combined with Qingyi decoction in the treatment of acute pancreatitis. Methods 120 patients with acute pancreatitis were chosen as observation subjects,and they were randomly divided into control group and observation group according to the digital table. The control group was treated with somatostatin,the observation group was treated with Qingyi decoction on the basis of control group. The clinical effects,improvements of clinical indicators and symptom,adverse reaction between the two groups were compared. Results The total effective rate of the observation group(100. 00%) was higher than that of the control group(90. 00%),the difference was statistically significant (χ2 =6. 316,P<0. 05). Before treatment, there were no statistically significant differences between the two groups in clinical indicators ( t =0. 047,0. 069, 0. 040,0. 279,0. 064,0. 101,0. 333,all P>0. 05). After treatment,the albumin of the observation group was higher than that of the control group,white blood cell count,APACHEⅡ score,C reactive protein,serum amylase,aspartate aminotransferase and lactate dehydrogenase were lower than those in the control group, the differences were statistically significant (t=14. 822,11. 654,15. 849,20. 000,10. 509,15. 213,15. 191,all P <0. 05). The blood amylase,abdominal pain relief,recovery of gastrointestinal function,urinary amylase,abdominal distension,ventilator weaning,abdominal tenderness disappeared and so on symptom improvement time of the observation group were earlier than those of control group,the hospital stay was shorter than that of the control group,the differences were statistically significant (t=10. 479,15. 855,8. 705,13. 064,8. 965,11. 783,11. 347,17. 069,7. 058,all P<0. 05). The incidence rate of adverse reactions in the control group was higher than that in the observation group(11. 67% vs. 1. 67%),the difference was statistically significant (χ2 = 4. 821, P < 0. 05 ). Conclusion Qingyi decoction combined with somatostatin can effectively improve the clinical symptom of acute pancreatitis,and with good clinical effects and safety.
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Objective To explore the effect of Wendan Decoction plus Saffron in the treatment of patients with endometriosis of uterus on serum matrix metalloproteninases-9(MMP-9).Methods 74 cases of endometriosis of uterus were selected and randomly divided into two groups, 37 cases in each group.The control group were treated with oral administration of Gestrinone Capsules, the research group were treated on the basis of the control group with Wendan Decoction plus Saffron treatment,two groups were treated for 24 weeks.The serum estradiol, MMP-9 and matrix metalloproteninases inhibitor-1(TIMP-1) were detected before and after treatment, the degree of pain and clinical efficacy were compared.Results Compared with before treatment, levels of estradiol(E2),progesterone(P) and luteotropic hormone(LH) decreased in two groups(P<0.01), the levels of MMP-9, MMP-9/TIMP-1 decreased(P<0.01), levels of TIMP-1 increased(P<0.01), levels of CA125 decreased(P <0.01), and the maximum diameter of pelvic mass line decreased(P<0.01), dysmenorrhea pelvic pain, dyspareunia and VAS score decreased(P<0.01).Compared with the control group, levels of E2, P and LH in the research group were lower(P<0.01), the levels of MMP-9, MMP-9/TIMP-1 were lower(P<0.01), levels of TIMP-1 were higher(P<0.01), levels of CA125 were lower(P<0.01), the maximum diameter of pelvic mass line were lower(P<0.01), dysmenorrhea and pelvic pain and VAS scores were lower(P<0.01), and the effective rate of research group was higher(P<0.05).Conclusion Wendan Decoction plus Saffron in the treatment of endometriosis of uterus was effective, and it can reduce endometriosis, dysmenorrhea and pelvic pain degree, reduce pelvic mass, and may reduce the level of serum MMP-9 related.
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Thirty-two articles from January 2000 to December 2014,with ankylosing spondylitis treated by acupuncture,moxibustion and medicines(Chinese medicine or western medicine) and statistically significant and effective results,were collected through Chinese National Knowledge Infrastructure(CNKI),WANFANG and VIP databases. It is concluded that there are various methods except medicines treating ankylosing spondylitis,including acupuncture,acupuncture combined with moxibustion,warm acupuncture,electroacupuncture,apitherapy,fire needle therapy,etc. Further studies are needed to be implemented so as to promote the combination therapy of acupuncture and medicine for ankylosing spondylitis,such as the standard cases of the combination therapy and mechanism.
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Objective To estimate the influence of curcumin on psoriasis-like lesions and proliferating cell nuclear antigen (PCNA) expression in guinea pigs,so as to investigate the therapeutic effect and mechanism of curcumin in psoriasis.Methods A model of psoriasis-like skin inflammation was established by applying propranolol 5％ cream (4 times a day for 3 weeks) to the dorsal skin of ears of 30 guinea pigs,which were then equally classified into 5 groups:model group receiving no treatment,observation group receiving no treatment but observation for 2 weeks,model control group treated with intragastric 25％ polyethylene glycol solution (1 rnl once a day) for 2 weeks,low-dose and high-dose curcumin groups treated with intragastric curcumin solution at 20 and 40 mg/kg per day respectively once a day for 2 weeks.Six guinea pigs receiving neither induction by propranolol nor treatment by curcumin or polyethylene glycol solution served as the normal control group.Skin specimens were harvested from the ears of guinea pigs in the normal control group after three weeks of breeding,in the model group immediately after the establishment of psoriasis-like model,in the observation group after 2 weeks of observation,and in the other 3 groups after 2 weeks of treatment.Subsequently,haematoxylin-eosin (HE) staining was conducted to observe histopathologic changes,and immunohistochemical assay to detect the expression of proliferating cell nuclear antigen (PCNA).Results Gross observation of the skin revealed that curcumin attenuated psoriasis-like skin manifestations in guinea pigs.There were significant differences in histopathologic scores (F=296.14,P＜ 0.01) and PCNA expression rate among the 6 groups (F =108.49,P ＜ 0.01).Least significant difference (LSD) test showed that both histopathologic scores and PCNA expression rate were significantly higher in the model group than in the normal control group (6.42 ± 0.49 vs.0.92 ± 0.20,63.17％ ± 5.47％ vs.20.83％ ± 2.99％,both P ＜ 0.01).After 2 weeks of treatment,both low-dose and high-dose curcumin groups showed significantly lower histopathologic scores (4.25 ± 0.27 and 1.75 ± 0.42 vs.6.42 ± 0.49,both P＜ 0.01) and PCNA expression rate (43.50％ ± 2.90％ and 25.50％ ± 3.74％ vs.63.17％ ± 5.47％,both P ＜ 0.01) compared with the model group.Conclusion Curcumin can attenuate pathological manifestations of psoriasis-like lesions,and downregulate PCNA expression in guinea pigs.
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Objective To establish the rabbit model with hepatic VX-2 tumor and to investigate the intake of folate-conjugated silica-coated gold nanorods (GNRs@SiO2-FA) in experimental rabbits. Methods Under CT-guidance, animal model with VX-2 liver cancer was established in 27 rabbits by using puncture inoculation method. CT scanning and sonography were employed to observe the tumor growth. After two weeks, the rabbits were randomly and equally divided into blank control group (n=9, injection of saline), portal vein injection group (n=9, injection of GNRs@SiO2-FA) and intra-tumoral injection group (n=9, injection of GNRs@SiO2-FA). Every three rabbits from each group were sacrificed each time at 24 h, 48 h and 72 h after the treatment. The tumor tissue and the major organs were collected and sent for pathological examination. The cellular uptake of GNRs@SiO2-FA was studied by confocal laser scanning microscopy. Results The rabbit model of VX-2 liver cancer was successfully established. CT and sonography examination indicated that the tumor was rich in blood supply. Confocal laser scanning microscopy revealed that GNRs@SiO2-FA could specifically bind with tumor cells within 24 hours after injection, then the GNRs@SiO2-FA entered into the tumor cells and gathered in the tumor cytoplasm. Conclusion GNRs@SiO2-FA has highly targeted effect on the liver cancer cells in experimental animals, which has very important application prospect in targeting hyperthermia therapy and in 125I seed implantation therapy.
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Objective:To investigate the affects of NOD2 on rapamycin (Rap)induced autophagy and on the proliferation and mi-gration of tongue squamous carcinoma SCC-1 5 cells.Methods:① Synthesized NOD2 over-expression plasmid and NOD2-shRNA were transfected into SCC-1 5 cells respectively.②Normal control SCC-1 5 cells,NOD2 over-expression cells and NOD2-shRNA cells were treated with Rap to induce autophagy.Then,the expression of LC3 and Beclin-1 was examined by Western blot.Cell pro-liferation was tested by MTT assay.Cell migration was assessed by wound healing assay.Results:①After Rap treatment,the expres-sion of protein LC3-Ⅱand Beclin-1 in NOD2 over-expression cells increased(P <0.05)and in NOD2-shRNA cells were suppressed (P <0.05).② Compared with control group,the proliferation and migration ability were decreased in NOD2 over-expression cells (P <0.05),but in NOD2-shRNA cells the proliferation and migration ability were increased(P <0.05).Conclusion:NOD2 can up-regulate the autophagy and suppress the proliferation and migration of tongue squamous SCC-1 5 cells.
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To observe the incidence of varicella vaccine after breaking through the case of varicella vaccine , immunization strategy ,popularized in the city.Inoculation Population living in the Binhai New Area of full age to 12 years old children in December,has been vaccinated or who have had chickenpox varicella vaccine except .Controls were four districts around the city girls without varicella in children.Methods: Implementation of vaccination for the target population.All vaccinees was observed from 42 days to 2 and a half years later ,the incidence of varicella break cases.In the observation group and the control group was observed in two groups of varicella vaccine protection rate calculation.Results: The gelatin free attenuated varicella vaccine breakthrough in 134 cases,the incidence rate was 0.35%;no gelatin attenuated varicella vaccine protection rate of 80.92%, with domestic and foreign reports consistent.Conclusion:After vaccination from 42 to 2 and a half years ,varicella vaccine can effectively protect children from the onset, while reducing the prevalence of children 's pain and the economic burden of the family.Varicella vaccine is still a breakthrough occurred ,therefore to consider two inoculations ,with further observation of two times after inoculation the body to produce antibody level and epidemiological protection effect.
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OBJECTIVE@#To investigate the effect of SOX2 on chemotherapy sensitivity of human laryngeal epithelial cells Hep-2.@*METHOD@#We designed and synthesized RNAis for silencing the expression of SOX2 in Hep-2 cells and selected the most effective RNAi by Western blot analysis. Then the recombinant plasmids of pGCsi-H1-SOX2 and pGCsi-H1-NC were constructed and transfected into Hep-2 cells to build cell lines of psiSOX2-Hep-2 and psiNC-Hep-2. CCK-8 assay had been used to test the sensitivity of Hep-2 cells to 5-FU and PTX after silencing SOX2 expression. Hoechst staining had been used to exam the changes of Hep-2 cells apoptosis treatment by 5-FU and PTX after silencing SOX2 expression. Furthermore, the changes of apoptosis-related genes expressions were detected by Western blotting.@*RESULT@#The cell lines of psiSOX2-Hep-2 and psiNC-Hep-2 were successfully established, and the expression of SOX2 protein was decreased 78% in psiSOX2-Hep-2 cells compared with psiNC-Hep-2 cells. After reducing SOX2 expression, the sensitivity of Hep-2 cells to 5-FU and PTX were increased and the IC50 values for 48 h were decreased to 8.12 μg/ml and 5.16 μg/ml. Meanwhile, the apoptosis rate and the expression of apoptotic gene Bax and cleaved caspase-3 expression were dramatically increased and anti-apoptotic genes survivin and Bcl-2 were significantly decreased in psiSOX2-Hep-2 cells compared with psiNC-Hep-2 cells.@*CONCLUSION@#Down-regulating the protein expression of SOX2 by RNAi will significantly enhance the sensitivity of human laryngeal epithelial cells Hep-2 to 5-FU and PTX.
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Humans , Apoptosis , Cell Line, Tumor , Cell Proliferation , Drug Resistance, Neoplasm , Epithelial Cells , Metabolism , Pathology , Fluorouracil , Pharmacology , Laryngeal Neoplasms , Metabolism , Pathology , RNA Interference , SOXB1 Transcription Factors , GeneticsABSTRACT
Objective:To explore the effects of NOD2 gene on the proliferation and apoptosis of tongue squamous cell carcinoma Tca8113 cells.Methods:NOD2 expression vector(NOD2-pEZ-M29)and NOD2-shRNA vector were established,then were trans-fected into Tca8113 cells respectively.Expressions of HBD-2 and NOD2 in the cells was detected by RT-PCR and Western blot.Cell proliferation was examined by MTT assay and apoptosis by flow cytometry at 48h post transfection.Results:Compared with the control group,the expression of NOD2 and HBD-2 in NOD2-pEZ-M29 transfection group was significantly higher and markedly lower in NOD2-shRNA group.The proliferation rate of Tca8113 cells was markedly lower in NOD2-pEZ-M29 transfection group and signifi-cantly higher in NOD2-shRNA group while the apoptosis rate was significantly higher in NOD2-pEZ-M29 transfection group and sig-nificantly lower in NOD2-shRNA group.Conclusion:In Tca8113 cells NOD2 expression was positively correlated with HBD-2 ex-pression.NOD2 gene may promote the apoptosis,inhibit the proliferation of Tca8113 cells.
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Objective To investigate the relationship between monocyte chemoattractant protein-1 (MCP-1) A-2518G single nucleotide polymorphism (SNPs) and susceptibility of epithelial ovarian cancer(EOC) in Chinese Han population of Hunan region.Methods MCP-1 A-2518G SNPs of the EOC were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis in 92 patients with EOC and 38 healthy women as control.Results MCP-1 A-2518G SNPs had AA,AG,and GG genotypes in cancer and control groups.The frequencies of AA,AG,and GG genotypes were not significantly different between cancer and control groups (AA:17.40％ and 15.79％ ; AG:44.56％ and 52.63％ ; and GG:38.04％ and 31.58％ ; P ＞0.05).Multivariate logistic regression analysis showed that there was no significant correlation between MCP-1 A-2518G polymorphism and EOC (P ＞0.05).Conclusions This present study suggested that MCP-1 A-2518G polymorphism should not be related to susceptibility of EOC in the Chinese Han population of Hunan region.
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BACKGROUND: There is an intimate temporal and spatial relationship between growth of primitive cardiac cells, septum transversum mesenchyme and liver development. The signal from primitive cardiac cells and septum transversum mesenchyme induces the ventral foregut endoderm cells specialize toward hepatocytes. While the septum transversum mesenchyme provides a suitable environment for forming the liver bud and promoting the growth and differentiation. However, the molecular mechanism of this induction is not yet delineated.OBJECTIVE: Using alpha-fetal protein (AFP), c-Met and cytokeratin (CK) 19 as markers of hepatic stem cells, the growth of early human embryo of 3-5 weeks and morphologic characteristic of hepatic stem cells were observed to demonstrate the characteristic and factors that affected the proliferation and differentiation of hepatic stem cell, which provided experimental evidence for basic research and clinical application of hepatic stem cells.DESIGN: An opening experiment was designed.SETTING: Department of Anatomy, Weifang Medical College.MATERIALS: The experiment was carried out at the Scientific Research Center of Chengdu Medical College between September 2004 and January 2005. Twenty cases fresh human embryos aged less than 2 months were collected with signed agreements of the pregnant women suffering from pregnancy termination with mifepristone. The samples were fixed with 40 g/L polymerisatum in 20 minutes and embedded routinely in paraffin, and then 5 μm thick series sections were continuously made. After hematoxylin-eosin staining, the embryonicage was determined under the microscope according to the length of embryos, the number of somites and the development of organs, which was referring to the Jirasek's staging standard of human embryo.METHODS: The immunohistochemical staining was conducted with SABC method on one of every ten sections, which were incubated overnight at 4 ℃ with polyclonal antibodies against hepatocyte growth factor (HGF),c-Met, insulin-like growth factor (IGF-Ⅰ), IGF-Ⅰ receptor (IGF-IR), transforming growth factor (TGFβ1), TGFβR1, TGFβR2 or monoclonal antibodies against proliferating cell nuclear antigen (PCNA), AFP and CK19.The following day, the sections were incubated for 2 hours at room temperature with biotinylated anti-mouse or anti-rabbit IgG and SABC liquid respectively, and then diaminobenzidine (DAB) was used to color them. The negative control was conducted with the phosphate buffer, then the sections were observed and photographed under light microscope.MAIN OUTCOME MEASUERS: ①the morphologic characteristic of human hepatic stem cells and immunohistochemical staining of markers②the expression of HGF, IGF-Ⅰ, TGFβ1 and their receptors on human embryonic livers of 3-5 weeks, primitive cardiac cells and septum transversum mesenchyme.RESULTS: ①The morphologic characteristic of human hepatic stem cells and immunohistochemical staining of markers: The hepatic bud formed at the end of 3rd week and migrated into the septum transversum mesenchyme to form the hepatic cords at the 4th week. The cells structuring the hepatic cords displayed the typical characteristic of immature cells. At the 5th week, the number of cells within the hepatic cords, the size of cell body,the cytoplasmic acidophilia all increased, whereas the basophilia of nuclei decreased. However the cellular forms were still homogeneous and displayed the typical characteristic of immature cells. The cells of hepatic cords were negative for PCNA response during 3rd-4th week but began to express positive at the 5th week, mainly in the nucleus and minority cellular cytoplasm showed weak positive. Most hepatic cells during 3rd-5th weeks were positive for AFP, c-Met and negative for CK19. The immunologic reaction depositors of AFP positive cells were located in the nuclei, cytoplasm and membrane of the hepatocytes, and c-Met presented mainly in the nuclei and the positive signal was weak in the cytoplasm. ②Expressions of HGF, IGF-Ⅰ, TGFβ1 and their receptors in the embryonic human liver, primitive heart and septum transversum mesenchyme: At the 4th week,the c-Met expressed only in all hepatocytes, whereas the other growth factors and their receptors were undetectable. At the 5th week, all the growth factors, except HGF, were expressed in the hepatocytes. The immunologic reaction depositors of TGFβ1, TGFβ1R1 and TGFβ1R2 were located in the cytoplasm and cell membrane. The positive response of IGF-Ⅰ and IGF-IR were present at nuclei, cytoplasm and cell membrane. At the 3rd-5th week, myocardial cells surrounding liver bud or hepatic cord and the septum transversum mesenchyme were positive for HGF, TGFβ1 and IGF-Ⅰ,with the signals were aggregated mainly in cytoplasm and minority nucei.CONCLUSION: ①It was at the end of 3rd week that part of endoderm cells in foregut ventral were specialized to hepatic stem cells. ②The undifferentiated hepatic stem cells could be drawn to develop to the liver stem cells with bi-directional differentiation potentials by using specific markers for studying human embryonic liver stem cells. According to the corresponding relation of embryonic age between human and rats, the time studying the rat hepatic stem cells could be calculated. ③HGF, IGF-Ⅰ,TGFβ1 and their receptors promoted the early development of human embryonic liver.
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BACKGROUND: The effective method to obtain neural stem cells through in vitro culture that deserves to pay more attention in experimental studies of stem cells.OBJECTIVE: To investigate the effective methods to culture neural stem cells in vitro using culture medium containing different growth factors.DESIGN: Single sample observation.SETTING: Laboratory of Cell Biology, College of Life Science, Sichuan University.MATERIALS: Ten SD rats which were pregnant for 14 days, DMEM/F12 1:1, basic fibroblasts growth factors, epidermal growth factor; nestin antibody IgG , β- microtubule protein Ⅲ antibody IgG , glial fibrillary acidic protein antibody IgG, biotin labeled second antibody and third antibody and fluorescinisothiocyate (FITC)-labeled second antibody were used in this experiment.METHODS: This experiment was carried out at the Laboratory of Cell Biology, College of Life Science, Sichuan University from 1999 to 2001. ① Brain tissue was taken out from the procerebrum of fetal rat, then primary neural stem cell clone was obtained through enzymatic digestion, mechanical treatment, centrifugation and culture. ② Neural stem cells were inoculated and cultured at 2×108 L-1 and divided into 4 groups with 6 bottles of cells in each group. DMEM/F12 (1:1 )culture medium containing 0.1 volume fraction of fetal bovine serum was added , serving as DMEM/F12 group; culture medium containing 20 μg/L basic fibroblast growth factor was added , serving as basic fibroblast growth factor group; culture medium containing 30 μg/L epidermal growth factor was added , serving as epidermal growth factor group; Culture medium containing basic fibroblast growth factors was added to culture for 2 hours, then culture medium containing epidermal growth factors was used for further culture, serving as basic fibroblast growth factor+ epidermal growth factor group. Culture flask was change after 24-hour culture; another 14 days later, primary clone number was counted under the microscope, and expressions of specially labeled albumen nidogen of stem cells were detected with immunohistochemistry.MAIN OUTCOME MEASURES: ① Primary clone of neural stem cells.② Clone culture result of unicell. ③ Induction and differentiation results.RESULTS: ① The cells isolated from the brain of fetal rats possess the ability to consecutively passage and form clone , and immunofluorescent staining showed cell nidogen expression positive in the cell sphere. ②Clone rate of neural stem cells was the highest (0.630%)using ordinal culture of basic fibroblast growth factor and epidermal growth factor. ③ The cultured neural stem cell clone can be induced and differentiated neurons and glinl cells.CONCLUSION: ① The results proved that the cultured and isolated cells are neural stem cells. ②Ordinal treatment of basic fibroblast growth factor and epidermal growth factor is an effective method to obtain neural stem cells.
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To study the morphological characteristics of hepatic stem cells and the expression of HGF, IGF-I, TGFbeta1 and their receptors in human embryonic livers at 3-5 weeks of gestation. The SABC immunohistochemical method with HE counterstaining was employed. We found that the hepatic bud formed at the end of the 3rd week. At the 4th week, the cells of hepatic bud migrated into the septum transversum mesenchyme and formed the hepatic cords. The hepatic cells at 3-4 weeks displayed the typical characteristics of immature cells: small size, a round or ovoid nucleus with dark color, scant cytoplasm with slight blue and a high ratio of nuclei/cytoplasm. They were positive for alpha-Fetoprotein (AFP), c-Met and negative for cytokertin 19 (CK19), and proliferating cell nuclear antigen (PCNA). At the 5th week, compared to those at the 4th week, the number of cells within the hepatic cords increased. But the cells at the 5th week were homogeneous and displayed the typical characteristic of immature cells. Those cells began to express PCNA at the 5th week. The hepatic cells at the 5th week were positive for insulin-like growth factor I (IGF-I), transforming growth factor beta1 (TGFbeta1) and their receptors, and were negative for hepatocyte growth factor (HGF), while HGF were positive in the cardiac cells and septum transversum mesenchyme. The results indicated that the cells of hepatic bud and cords were the hepatic stem cells. The difference of morphology and proteins expression at 3-5 weeks of gestation inferred that those stem cells belong to different developmental stage. AFP and c-Met were the markers of hepatic stem cells at the early stage of human embryo. HGF, IGF-I, TGFbeta1 and their receptors may involve in regulating the development of early embryonic human liver.
Subject(s)
Humans , Embryo, Mammalian , Gestational Age , Hepatocyte Growth Factor , Genetics , Insulin-Like Growth Factor I , Genetics , Liver , Cell Biology , Metabolism , Proto-Oncogene Proteins c-met , Genetics , Receptor, IGF Type 1 , Genetics , Stem Cells , Cell Biology , Transforming Growth Factor beta , Genetics , Transforming Growth Factor beta1ABSTRACT
Objective To isolate the bone marrow mesenchymal stem cells(MSCs) from the GFP-expressing mouse, and to study the osteoblastic differentation of the cells. Methods MSCs were isolated by density gradient centrifugation, then the clutrued cells were induced to osteoblastic differentiation using the conditional medium. We detectd the expression of GFP and MSCs differentiation into osteoblasts by histochemistry and immunochemistry. Results The MSCs maintained the expression of GFP during expanded and induced process. After induced for 10 days, lots of alkaline phosphatase and osteoclcin staining positive cells were observed.Conclusion The MSCs of GFP-expressing mouse were successfully isolated and differentiated into ostoblasts. It may be valuable for tracing the seeding cells in tissue engineering bone.
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Objective To study the distribution of insulin-like Growth Factors (IGFs) in liver、 spleen、kidney、thymus and supraadrenal glands of fetus from 16 to 24 weeks.Method Using ABC immunohistochemistry. Results IGF-I positive cells were found in these organs during this period of fetal life, but the differences of number and reaction intensity of IGF-I positive cells appeared in different organs and individuals.Conclusion These organs tissues can express IGF-I during fetal development and play paracrine and/or autocrine role in cellular proliferation and differentiation of these fetal tissue.
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Purpose To study the proteins expression of genes related to apoptosis of retinal cells in development of human fetus． Methods Fifty cases of retinas of human fetus aged from 12 to 38 weeks were collected and paraffin embedded sections were made． Immunohistochemical method was used． Results Fas protein was expressed by cells of ganglion cell layer, inner and outer nuclear later, which were just formed on 16th week． It was not expressed until 38th week, Fas(+) staining appeared in layers of retina． Fas-L(+) staining was detected in cells of layers of retina on 26th week and the positive staining located in ganglion cell layer on 32th week． Neuronal fiber layer was Fas-L positive． Bax positive staining was detected on 8th week． Bax positive nucleus were observed mainly in GCL and ONL on 16th week． It was in INL on 24th week and in Müller cells inner terminates on 26th week． After this time, all cells of retina were bax immune negative staining． Bcl-2(+) staining appeared in differentiating neuroblastic layer on 16th week． Beginning on 24th week, bcl-2 (+) staining was observed in glial cells of GCL and inner terminates of Müller cell． Conclusion Apoptosis of developing retinal cell may be Fas/Fas-L independent and bax may be involved in apoptosis of the cells．