The role of poloxamer 188 for cord blood mononuclear cells into megakaryocytes cultivation and induction in three-dimensional WAVE Bioreactor / 中华血液学杂志

Objective: To observe the effect of poloxamer 188 (P188) on megakaryocyte cultivation and induction from cord blood mononuclear cells in order to obtain more megakaryocyte progenitor cells (MPC). Methods: The cord blood mononuclear cells were isolated and inoculated in cell culture bag or cell culture flask respectively. The WIGGENS shaker and cell culture bags were used to mimick WAVE Bioreactor for three-dimensional (3D) cell culture, and the P188 was added to induction medium, The cells were detected for morphology, surface marker, viability, and number on day 14. Results: In the two-dimensional (2D) culture, CD41(+), CD41(+)/CD61(+), CD61(+) megakaryocytic numbers increased significantly after adding P188 (all P<0.01). And in the 3D culture of adding P188, the cell volume became larger and the nuclear shape was irregular, the cytoplasm appeared magenta granules, and the megakaryocyte cells became more mature. By 3D culture, the expression of CD41/CD61 was (36.30±1.27)% vs (23.95±1.34)%, hence the differentiation for MPC was significantly higher than that in the 2D group (P<0.01). Furthermore, adding P188 in 3D culture resulted in highest differentiation efficiency for MPC [(59.45±1.20)%]. There were no significantly differences in terms of cell viability and cell number among 3D culture containing P188, 2D and 3D culture groups (all P>0.05). Conclusion: 3D culture was beneficial for the differentiation of MPC, but the cell viability was lower than 2D group; However, the satisfied cell growth and better induction efficiency were obtained by adding of P188, which might provide a new method of megakaryocytes production for clinical application.

Mediating effect of loneliness on relationship between interpersonal adaptation and mobile phone addiction in college students / 中国心理卫生杂志

Objective:To explore the mediating effect of loneliness on the relationship between interpersonal adaptation and mobile phone addiction in college students.Methods:Totally 368 college students (176 males and 192 females )were surveyed with the College Student Adaptability (CSAI),University of Califomiaat Los Angels (UCLA)and Mobile Phone Addiction Index (MPAI).Then AMOS18.0 was used to establish structural equation modeling (SEM)among interpersonal adaptation,loneliness and mobile phone addiction,and test the mediating effect of loneliness on the relationship between interpersonal adaptation and mobile phone addiction in college students.Results:The MPAI scores were significantly higher in the low interpersonal adaptation group (the score of CSAI less than 21 )than in the high interpersonal adaptation group (the score of CSAI higher than 32)(P<0.001).The scores of interpersonal adaptation were negatively correlated with scores of loneliness and mobilephone addiction (r =-0.71,-0.25,P﹤0.01).The scores of loneliness were positively correlated with scores ofmobile phone addiction (r =0.32,P ﹤ 0.01).College students'interpersonal adaptation did not have significantnegative predictability for mobile phone addiction (β=﹣0.08,P =0.32),but it had significant negative predictabilityfor loneliness (β=﹣0.71,P <0.001).Loneliness had significant positive predictability for mobile phone addiction(β=0.29,P <0.001).Conclusion:The results show loneliness completely plays a mediating effect betweeninterpersonal adaptation and mobile phone addiction.

High-frequency Oscillations and the Seizure Onset Zones in Neocortical Epilepsy / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>To study the characters of high-frequency oscillations (HFOs) in the seizure onset zones (SOZ) and the nonseizure onset zones (NSOZ) in the electrocorticography (ECoG) of patients with neocortical epilepsy.</p><p><b>METHODS</b>Only patients with neocortical epilepsy who were seizure-free after surgery as determined with ECoG were included. We selected patients with normal magnetic resonance imaging before surgery in order to avoid the influence of HFOs by other lesions. Three minutes preictal and 10 min interictal ECoG as recorded in 39 channels in the SOZ and 256 channels in the NSOZ were analyzed. Ripples and fast ripples (FRs) were analyzed by Advanced Source Analysis software (ASA, The Netherlands). Average duration of HFOs was analyzed in SOZ and NSOZ separately.</p><p><b>RESULTS</b>For ripples, the permillage time occupied by HFOs was 0.83 in NSOZ and 1.17 in SOZ during the interictal period. During preictal period, they were 2.02 in NSOZ and 7.93 in SOZ. For FRs, the permillage time occupied by HFOs was 0.02 in NSOZ and 0.42 in SOZ during the interictal period. During preictal period, they were 0.03 in NSOZ and 2 in SOZ.</p><p><b>CONCLUSIONS</b>High-frequency oscillations are linked to SOZ in neocortical epilepsy. Our study demonstrates the prevalent occurrence of HFOs in SOZ. More and more burst of HFOs, especially FRs, means the onset of seizures.</p>

Reliability and validity of Connor-Davidson resilience scale in Chinese college students / 中华行为医学与脑科学杂志

Objective To examine the reliability and validity of Chinese revision of connor-davidson resilience scale(CD-RISC) in Chinese college students.Methods 1610 college students were tested with CD-RISC.783 of them took a more test by BFI and SCL-90 at the same time.After three weeks,83 students were retested by CD-RISC.Results (1) The results of confirmatory factor analysis (x2/df =6.632,RMSEA =0.059,GFI =0.934,AGFI =0.915,CFI =0.927,NFI =0.915) indicated that the three-factor model reasonably fitted Chinese college students.(2) The Cronabach α coefficients of the CD-RISC and three factors called adaptability,tenacity and autonomy were 0.914,0.865,0.784,0.767 respectively; the mean inter-item correlation coefficients of them were 0.364,0.417,0.425,0.398 respectively; the Guttman split-half coefficients were 0.888,0.843,0.707,0.650 respectively; the retest-reliability coefficients were 0.856,0.742,0.777,0.747.(3) The scores of the total scale of CDRISC and three factors were significantly correlated to BFI and SCL-90(P＜0.01).There were significant differences between the low-resilient group and the high-resilient group in every index of BFI and SCL-90.(4) There were significant differences between male and female students in CD-RISC and factor tenacity and autonomy(male:2.77±0.60,2.70-±0.74,2.65±0.68;female:Z70±0.53,2.62±0.67,2.54±0.62; P＜0.05).Conclusion The Chinese version of CD-RISC is a reliable and valid method for assessing resilience in Chinese college students.

Differentiation potential of CD41⁺ cells derived from the mouse aorta-gonad-mesonephros region, yolk sac and embryonic circulating blood / 中华血液学杂志

<p><b>OBJECTIVE</b>To compare the differentiation ability difference of hematopoietic, mesenchymal and endothelial potential between CD41⁺ cells derived from the mouse aorta-gonadmesonephros (AGM) region, yolk sac (YS) and embryonic circulating blood (CB).</p><p><b>METHODS</b>CD41⁺ cells were sorted from AGM, YS and CB. The CD45 and c-kit expression were studied in CD41⁺ cells by flow cytometry. IL-3 and bone morphogenetic protein 4 (BMP-4) treatment together with semi solid culture were used to assess hematopoietic potential difference of CD41⁺ cells. Immunofluorescence staining of α-SMA was used to assess mesenchymal potential difference. The endothelial cell induction system was used to assess endothelial potential difference.</p><p><b>RESULTS</b>The proportions of CD45+ cells in CD41⁺ population were 51.9% (AGM), 45.8% (YS) and 22.2% (CB), respectively, while those of c-kit⁺ cells were 40.0% (AGM), 39.6% (YS) and 36.2% (CB), respectively. After stimulated by IL-3 factor, the number of total colonies increased in all three groups-derived CD41⁺ cells compared to that of unstimulated group[(14.1±1.9) vs (1.2±0.2), (32.4±1.1) vs (18.4±2.2) and (41.8±0.9) vs (10.4±1.8)], (P<0.01). After stimulated by BMP-4 factor, compared to unstimulated group, CFU-Mix colony number in CD41⁺ cells from AGM region and YS were significantly decreased[(0.5±0.6) vs (3.2±0.8), (1.3±0.7) vs (7.4±1.7)](P<0.01), but there was no difference in CB group[(2.5±0.5) vs (3.9±1.5)](P>0.01). The mesenchymal marker α-SMA was highly expressed in CD41⁺ cells from AGM region and YS, but lowly expressed in CD41⁺ cells from CB.</p><p><b>CONCLUSION</b>There are some differences between CD41⁺ cells in AGM region, YS and CB on hematopoietic cell surface marker expression, hematopoietic colony formation with IL-3 and BMP-4 stimulation.</p>

Factorial structure of connor-davidson resilience scale in Chinese college students / 中华行为医学与脑科学杂志

ObjectiveTo explore the factorial structure of Chinese revision of connor-davidson resilience scale (CD-RISC) in Chinese college students.MethodsA total of 1534 college students were recruited for this study.After item analysis,half of the sample was used for exploratory factor analysis and the other for confirmatory factor analysis.ResultsThe Chinese revision of CD-RISC contained 19 items.Exploratory factor analysis showed that three factors were better:adaptability,tenacity and autonomy.And the results of confirmatory factor analysis ( x2/df =3.83,RMSEA =0.06,GFI =0.92,AGFI =0.90,CFI =0.92,NFI =0.89) indicated that this model provided a reasonable good fit for Chinese college students.ConclusionThis study indicate that the three-factor model of CD-RISC is adaptable to Chinese college students.

Application of melanocytes and bone marrow mesenchymal stem cells in tissue engineered skin construction / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the method of constructing tissue-engineered skin using melanocytes and bone marrow mesenchymal stem cells (BMSCs) in vivo.</p><p><b>METHODS</b>Melanocytes were isolated from human foreskin. BMSCs were isolated from human bone marrow. Both of them were co-cultured at a ratio of 1:10, and then were implanted into the collagen membrane to construct the tissue-engineered skin, which was applied for wound repair in nude mice. The effectiveness of wound repair and the distribution of melanocytes were evaluated by morphological observation, in vivo 4,6-diamidino-2-phenylindole, dihydrochloride (DAPI) fluorescent staining tracing, HE staining, S-100 immunohistochemistry, and transmission electron microscopy.</p><p><b>RESULTS</b>The wounds were satisfactorily repaired among the nude mice. The melanocytes were distributed in the skin with normal structure, as confirmed by DAPI fluorescent staining tracing, HE staining, S-100 immunohistochemistry, and transmission electron microscopy.</p><p><b>CONCLUSION</b>Melanocytes and BMSCs, after proper in vitro culture at an appropriate ratio, can construct the tissue-engineered skin with I type collagen membrane.</p>

Observation of bone marrow-derived mesenchymal stem cells targeting glioma with in vivo bioluminescence imaging / 中华神经医学杂志

Objective To explore the intracranial distribution of bone marrow-derived mesenchymal stem cells (BMSCs) and the ability of BMSCs shifting to glioma tissue.Methods We isolated BMSCs from the rats and constructed a BMSCsRL model that can stably express Renilla luciferase (RL).And 9L glioma cells marked with PKH26 were implanted into the brain parenchyma of Fischer rat using stereotactic surgery;7 d after that, the BMSCsRL was implanted into the contralateral brain parenchyma.The intracranial distribution of BMSCsRL was detected by using Xenogen bioluminescance imaging (BLI);at the same time,the migration of BMSCsRL into the glioma tissue was observed using Transwell plates.Results Phenotypical properties of the isolated BMSCs were CD90 and CD44 positive.BMSCs could be targeted to glioma tissue.In vivo BLI showed that the BMSCs shifted to the glioma tissue 0,7 and 14 d after transplantation and the junction area between tumor tissue and normal tissue was much more obvious than the other areas.Conclusion These results confirm the migratory capability of BMSCs over considerable distances, suggesting that BMSCs can be used as a delivery vehicle for targeted therapy of glioma.

Application of Aidi injection (艾迪注射液) in the bronchial artery infused neo-adjuvant chemotherapy for stage III A non-small cell lung cancer before surgical operation / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the effect of Aidi Injection (艾迪注射液,ADI) applied in the bronchial artery, applied in the bronchial artery infused (BAI) neo-adjuvant chemotherapy for stage III A non-small cell lung cancer (NSCLC) before surgical operation.</p><p><b>METHODS</b>The 60 patients with NSCLC stage III A underwent two courses BAI chemotherapy before tumor incision were assigned to two groups, the treatment and the control groups, using a random number table, 30 in each group. ADI (100 mL) was given to the patients in the treatment group by adding into 500 mL of 5% glucose injection for intravenous dripping once daily, starting from 3 days before each course of chemotherapy, and it lasted for 14 successive days, so a total of 28 days of administration was completed. The therapeutic effectiveness and the adverse reaction that occurred were observed, and the levels of T-lymphocyte subsets, natural killer cell activity, and interleukin-2 in peripheral blood were measured before and after the treatment.</p><p><b>RESULTS</b>The effective rate in the treatment group was higher than that in the control group (70.0% vs. 56.7%, P<0.05). Moreover, as compared with the control group, the adverse reaction that occurred in the treatment group was less and mild, especially in terms of bone marrow suppression and liver function damage (P<0.05). Cellular immune function was suppressed in NSCLC patients, but after treatment, it ameliorated significantly in the treatment group, showing significant difference as compared with that in the control group (P<0.05).</p><p><b>CONCLUSION</b>ADI was an ideal auxiliary drug for the patients in stage III A NSCLC received BAI neo-chemotherapy before surgical operation; it could enhance the effectiveness of chemotherapy, ameliorate the adverse reaction and elevate patients' cellular immune function; therefore, it is worthy for spreading in clinical practice.</p>

Effects of electroacupuncture on the expression of nestin in endogenous neural stem cell in hippocampus in rats with focal cerebral ischemia-reperfusion / 中国针灸

<p><b>OBJECTIVE</b>To explore the mechanism of electroacupuncture for treating focal cerebral ischemia-reperfusion injury.</p><p><b>METHODS</b>Seventy-five Wistar rats were randomly divided into a control group, a model group and an electroacupuncture group, 25 cases in each group. The model of focal cerebral ischemia-reperfusion was established by inserting nylon thread into the internal carotid artery except the control group which was only separated of the carotid artery without occlusion. Electroacupuncture group was treated with electroacupuncture at "Baihui (GV 20)" and "Dazhui (GV 14)" and the other groups without electroacupuncture treatment. The number of nestin positive cells expression at 1st, 3rd, 7th, 14th and 21st days after focal cerebral ischemia reperfusion was observed by use of immunohistochemistry method.</p><p><b>RESULTS</b>The number of nestin positive cells in electroacupuncture group at ischemia side was significantly more than that in the model group at 3rd, 7th, 14th and 21st days (P < 0.05, P < 0.01), and at contralateral ischemia side, the number of nestin positive cells in the electroacupuncture group was significantly more than that in the model group only at 7th day (P < 0.05).</p><p><b>CONCLUSION</b>Electroacupuncture at "Baihui (GV 20)" and "Dazhui (GV 14)" in rats can increase the number of nestin positive cells in hippocampus after focal cerebral ischemia-reperfusion, which may be one of the important mechanisms of electroacupuncture in treating acute cerebral ischemic diseases.</p>

Erythropoietin gene-modified conditioned medium of human mesenchymal cells promotes hematopoietic development from human embryonic stem cells / 中国实验血液学杂志

The study was aimed to investigate the effect of deriving hematopoietic cells from human embryonic stem cells (hESCs) by the erythropoietin gene-modified conditioned medium of human mesenchymal cells. The mesenchymal stem cells (MSCs) steadily expressing EPO were established by lentiviral system. The expression of exogenous EPO was detected by RT-PCR and Western blot. After suspension culture, hESCs developed into embryonic bodies (EBs). Then the EB cells were cultured in conditional medium. The hESCs-derived hematopoietic cells were analyzed by immunofluorescence, CFU assay and RT-PCR. The results indicated that the exogenous EPO successfully expressed in the EPO transfected MSCs (EPO/MSCs). The supernatant from EPO/MSCs increased CD34(+) cell population and the expression of globin, and enhanced colony forming unit incidence. These effects were obviously higher than that of control. It is concluded that the EPO gene-modified conditioned medium of human mesenchymal cells can induce the hESCs to differentiate into hematopoietic cells.

Induction of hepatic specification of human adipose-derived stem cells (hADSCs) in vitro / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To induce hepatic differentiation of human adipose-derived stem cells (hADSCs) in vitro.</p><p><b>METHODS</b>hADSCs were isolated from human adipose tissue and treated with improved hepatic medium containing HGF, bFGF and FGF4. After 7 days of culture, OSM was added to the culture media. Cell growth during hepatic differentiation was evaluated by CCK8 assay. Morphology of differentiation was examined under light microscope. Liver specific genes and proteins were detected by RT-PCR analysis and immunohistochemical staining, respectively. And functional characteristics of hepatocytes were also examined.</p><p><b>RESULTS</b>The number of hADSCs cultured in the improved hepatic media was increased significantly in comparison to hADSCs cultured in control media from 5 days to 21 days (t=6.59, 8.69, 15.94 and 24.64, respectively, P<0.05). The hADSCs-derived hepatocyte-like cells exhibited hepatocyte morphology, expressed hepatocyte markers, possessed hepatocyte-specific activities, such as uptake and excretion of indocyanine green, glycogen storage and albumin production.</p><p><b>CONCLUSION</b>hADSCs can be induced into hepatocyte-like cells in this differentiation system. And this differentiation system promoted the growth of hADSCs.</p>

Effect of doxorubicin induced GM-CSF gene expression regulated by Egr-1 promoter on recovery of hematopoietic function in bearing-melanoma mice / 中国实验血液学杂志

This study was purposed to investigate the effect of hematopoietic growth factor expression regulated by Egr-1 promoter on the recovery of hematopoietic function in bearing-melanoma mice after chemotherapy with doxorubicin (ADM). The human GM-CSF cDNA and enhanced green fluorescence protein (GFP) cDNA were linked together with internal ribozyme entry site (IRES) and then were inserted into the expression vector pCI-neo under control of the Egr-1 promoter (Egr-EG). This vector was transduced into human bone marrow stromal cell lines HFCL by lipofectamine and was transfused in severe combined immunodeficiency (SCID) mice. The experimental mice were randomly divided into 4 groups (6 mice in each group): (1) HFCL/EG + ADM group in which the HFCL/EG cells were transplanted intravenously in SCID mice bearing melanoma, ADM was given intraperitoneally after 3 days; (2) HFCL + ADM group in which the HFCL cells were transplanted intravenously, ADM was given intraperitoneally after 3 days; (3) HPCL/EG group in which HFCL/EG cells were transplanted alone; (4) HFCL group in which HFCL cells were transplanted alone. The dynamic change of peripheral blood picture was assayed by hemocytometer; the eGFP(+) human stromal cells were detected by flow cytometry; the expression of GM-CSF mRNA and protein were determined by RT-PCR and Western blot respectively. The results indicated that as compared with HFCL/EG and HFCL groups, the leukocyte count in HFCL/EG + ADM group decreased, but decrease level was weaker than that in HFCL + ADM group, meanwhile the recovery of leukocyte count was earlier than that in HFCL + ADM group. The CFU-GM amount between 4 groups showed no significant difference. The detection results showed that the inhibitory rate of tumor was related to chemotherapy, but not to expression of exogenous gene; the eGFP(+) stromal cells existed in bone marrow of mice treated with ADM. The RT-PCR and Western blot assays revealed enhancement of GM-CSF mRNA and protein. It is concluded that the ADM-inducible GM-CSF gene therapy regulated by Egr-1 promoter may promote the hematopoietic recovery after chemotherapy.

Experimental study of transplanting human mesenchymal stem cells via portal vein to treat acute liver injury of mice induced with acetaminophen / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To evaluate the effects of human mesenchymal stem cells (hMSCs) transplantation via portal vein to treat acute liver injury in mice induced with acetaminophen.</p><p><b>METHODS</b>A model of acute liver injury was established by acetaminophen gavage with a dose of 500 mg/kg. Twenty severe combined immune deficient mice (SCID mice) were randomly divided into 2 groups; one with hMSCs transplantation via their portal veins, the other group served as controls and only saline was infused into their veins. Liver function tests, fluorescein staining and reticular fiber staining of liver histological preparations and fluorescence- and light-microscopy were applied to observe the biochemical and pathological changes in the mice before and after the transplantation of hMSCs.</p><p><b>RESULTS</b>Liver function of the hMSCs group was significantly better than that of the controls (P less than 0.05). Fluorescence microscopy revealed that the hMSCs appeared in the areas of the periportal veins at first and then extended to the central vein areas; the reticular fiber staining indicated that hMSCs could repair the architecture of the hepatic acini. No prominent fibrosis and pseudolobules were found.</p><p><b>CONCLUSIONS</b>hMSCs transplantation via portal vein to SCID mice with acute liver injury induced by acetaminophen can improve their liver function effectively; hMSCs growth in their livers and acinus reconstruction can be affected. We think it is a good method to treat acute liver injury.</p>

Regulatory effect of Egr-1 promoter sequences induced by doxorubicin in transcriptional targeting on expression of GM-CSF gene / 中国实验血液学杂志

In order to explore the regulatory effects of Egr-1 promoter sequences induced by doxorubicin (ADM) in transcriptional targeting on the expression of hematopoietic growth factor genes. The human GM-CSF cDNA and enhanced green fluorescent protein (eGFP) cDNA were linked together with internal ribozyme entry site (IRES) and then inserted into the expression vector pCIneo under control of the Egr-1 promoter (Egr-EG). The vector was transferred into human bone marrow stromal cell line HFCL by lipofectin. The transfected cell clones (HFCL/EG) were selected by the addition of G418. The cells were exposed to the clinically important anticancer agent doxorubicin. The activity of eGFP in HFCL/EG cells was detected by flow cytometry. The amounts of GM-CSF in HFCL/EG postchemotherapy were confirmed with ELISA. The effect of GM-CSF in HFCL/EG cultural supernatants on expansion of CFU-GM derived from cord blood was also studied. The effect of N-acetylcysteine (a free radical scavenger) on GM-CSF production following exposure to ADM was examined. The results indicated that the activity of eGFP and the amounts of secreted GM-CSF in HFCL/EG cells exposed to ADM increased as compared to non-ADM group. The effect of GM-CSF in HFCL/EG cultural supernatants on expansion of CFU-GM was significantly higher than that of non-ADM group. N-acetylcysteine significantly decreased the concentration of GM-CSF produced by HFCL/EG treated with ADM. It is concluded that these in vitro data provide an experimental basis for the use of gene therapy of hematopoietic growth factor gene regulated by Egr-1 promoter to protect hematopoiesis from ADM-injury.

FRIL maintains quiescence of hematopoietic stem cells through regulation of cell cycle related factors / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the mechanism of Flt3 receptor-interacting lectin (FRIL) maintains quiescence of hematopoietic stem cells (HSCs) in vitro.</p><p><b>METHODS</b>Cord blood CD34+ cells were cultured in suspension medium supplemented with or without FRIL and FL. Cells were collected at different time points and the expression of some cell cycle regulators, especially those involved in G0/G1 phase regulation were detected on mRNA and protein level.</p><p><b>RESULTS</b>The expressions of G0/G1 phase related cyclins or CDKs were undetectable in the newly isolated CD34+ cells, expressions of Cyclin D3, CDK6 and P27 were the lowest in FRIL cultured group after 3d's culture (FRIL group: 483 +/- 63, 553 +/- 39, 0.312 +/- 0.030; FL group: 2437 +/- 52, 3209 +/- 98, 0.787 +/- 0.024; BLANK: 914 +/- 105, 1497 +/- 55, 0.616 +/- 0.029, respectively), but the expression of P53 was the highest in FRIL group (FRIL group: 4.476 +/- 0.159; FL group: 0.581 +/- 0.099, BLANK: 2.167 +/- 0.114). The expression of positive regulators of cell cycle in FRIL group were the same as that of FL group and blank group or lower.</p><p><b>CONCLUSION</b>FRIL preserves HSCs effectively in vitro through the mechanisms of down-regulation of cyclin D3 and CDK6 and activation of P53. P27 is mostly involved in the differentiation of HSCs.</p>

Differentiation of bone marrow derived Thy-1+ beta2M- cells into liver cells in AA induced liver injury micro-environment / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the differentiation of bone marrow derived Thy-1+ beta2M- cells (BDTCs) into liver cells in allyl alcohol (AA) induced liver injury micro-environment.</p><p><b>METHODS</b>BDTCs of male F344 rats were isolated by two-step magnetic separation system (MACS) technique, and infused intraportally into female recipients after labeling with PKH26. Thirty recipients were divided randomly into 3 groups: (1) AA-injured liver + BDTCs infusion, (2) normal liver + BDTCs infusion and (3) AA-injured liver + NS infusion (control). Blood biochemical examination, fluorescence labeled cellular localization, Y-chromosome sry gene in-situ hybridization and immunohistochemistry were carried out to evaluate BDTCs distribution, differentiation and proliferation in recipients's livers after different intervals.</p><p><b>RESULTS</b>Fluoromicroscopy and in situ hybridization suggested that BDTCs of donors were interspersed in pieces and cords among the necro-periportals induced by AA; immunohistochemistry indicated that those implanted cells expressed OV-6, AFP, CK19 and albumin successively, while positive cells were hardly seen in the normal liver + BDTCs infusion group. Compared with the controls, the blood biochemical restitution was more rapid in group (1), (9.8 d +/- 3.1 d vs. 13.7 d +/- 4.2 d).</p><p><b>CONCLUSION</b>The injury micro-environment induced by AA facilitates BDTCs integration with hepatic cell plates and differentiation into mature liver cells. BDTCs differentiation into liver cells might accelerate endogenous liver cell regeneration and reparation.</p>

Differentiation of rat bone marrow derived Thy-1+ beta2M- cells into hepatocytes / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the differentiation of bone marrow derived Thy-1(+)beta(2)M(-) cells (BDTC) into mature and functional liver cells and its mechanism.</p><p><b>METHODS</b>BDTC were cocultured with allyl alcohol (AA)-injured hepatocytes and cultured alone in conditional medium containing HGF and bFGF, respectively. BDTC morphologic transformation was observed with phase-contrast and electron-microscopy. Hepatocyte-specific gene expression in cultured BDTC was identified by immunocytochemistry and reverse transcription-polymerase chain reaction (RT-PCR). Indocyanine green (ICG) ingestion/excretion and urea, albumin production were carried out to evaluate hepatocyte-related function.</p><p><b>RESULTS</b>Some BDTC derived hepatocyte-like cells with high nuclear to cytoplasmic ratio containing mono- or multi-nuclei and abundant mitochondria, endoplasmic reticulum and glycogenic granules appeared after 7-day culture in both the two culture systems. These cells expressed hepatocyte-specific genes (AFP, OV-6, CK18, etc.), and possessed functions of ICG uptake, albumin production and ammonium metabolism.</p><p><b>CONCLUSION</b>Rat BDTCs could differentiate into mature and functional liver cells in special stimulation systems. Moreover, these differentiations were realized by "transdifferentiation", and might dispense with "cell fusion".</p>

Regulatory role of HTm4 gene in hematopoietic cell cycle / 生理学报

Cell cycle progression is tightly regulated in hematopoietic stem cells. The cycle state decides cells' fates, which includes self-renewal, proliferation and differentiation. Proper cell cycle regulation is a pivotal element for the maintenance of hematopoiesis homeostasis. HTm4 is a newly identified specific cell cycle regulator of the hematopoietic cell. Through interacting with KAP-CDK2 complex, it arrests cells in G(0)/G(1) phase. K562 is a human chronic myelogenous leukemia cell; it could be induced to megakaryoblast by phorbol 12-myristate 13-acetate (PMA). Such differentiation must be associated with cell cycle change. To further clarify HTm4's function in hematopoietic cell cycle regulation, K562 cells were treated with PMA. Cell cycle change was analysed using flow cytometric system. And during the induction process gene expression of HTm4 as well as CycleE and CDK2, which are responsible for G(1) to S transition, were analysed using semi-quantitative RT-PCR. The C-terminal domain of HTm4 protein has been shown to be important for HTm4's binding with KAP-CDK2 complex. To determine its impact on HTm4's function, HTm4 and C-terminal truncated HTm4 (HTm4-ct) were transfected into K562 cells using Tet-Off regulation expression system. Their influence on cell cycle was observed. The results showed that PMA induced both expansion and differentiation of K562 cells as measured by cell number count and NBT staining respectively. During PMA treatment, G(0)/G(1) cell proportion and HTm4 expression displayed coordinated change, which suggested that HTm4 might drive K562 cells out of cell cycle but was not involved in the quiescence maintenance. Additionally, transfection of HTm4 caused G(0)/G(1) arrest in K562 cells, while transfection of HTm4-ct did not. It is therefore suggested that the C-terminal domain is important for the function of HTm4 in cell cycle regulation.

Sustaining effect of gene-transferring hepatic stellate cell strain CFSC/HGF on hepatocytes development / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the sustaining effects of gene-transferring hepatic stellate cell strain CFSC/HGF on the development of hepatocytes.</p><p><b>METHODS</b>A CFSC/HGF strain, expressing HGF steadily and effectively was established by recombined retroviral vector pMSCV-HGF infection. Morphology and ultra structure of hepatocytes, albumin and urea production, as well as ICG uptake and excretion were studied continuously following the hepatocytes cultured on the CFSC/HGF feeder layers. Parallel group of collagen-dependent hepatocytes culturing and hepatocytes culturing on CFSC were also conducted. Semi-quantitative RT-PCR analysis was made to evaluate the expression of HGF receptor c-Met.</p><p><b>RESULTS</b>The hepatocytes cocultured on CFSC/HGF feeder layers had a higher survival rate, and the functions of albumin secretion and urea syntheses and ICG uptake and excretion, were superior to the other two culture methods. The result of RT-PCR indicated that the c-Met expressed on the CFSC/HGF coculturing hepatocytes was up-regulated 2.23 times.</p><p><b>CONCLUSION</b>Gene-transferring hepatic stellate cell strain CFSC/HGF exhibited a remarkable sustaining effect on the hepatocytes development. The up-regulation of c-Met expressed on the surfaces of the hepatocytes induced by CFSC/HGF might play some part in this function.</p>