BACKGROUND: At present, studies show that a kind of stem cell community which in many kinds of organizations can differentiate into tissue cells of different embryonic layers; but those are different from embryonic stem cells, embryonic stem cell will lose the part differentiation potential gradually during the development of pregnancy, and will present some special phenotypes or the molecular markers, as CD105 and so on, will cell it postembryonic pluripotent stem cells.OBJECTIVE: To study the isolation of postembryonic pluripotent stem cells from fetal bone marrow, proliferative culture in vitro, induction and differentiation; transplantation to the liver of SCID mice with hepatic failure, and detect therapy effects.DESIGN, TIME AND SETTING: Cell observation and animal randomization experiment which was completed in the Ministry of Health of Cell Engineering Technology Research Center, Tianjin Third Central Hospital from March 2003 to March 2005.MATERIALS: The postembryonic pluripotent stem cells were extracted from thighbone and shinbone of 22-week old fetuses under sterile circumstance. Adult female SCID mice ware regarded as the recipients. CD105 immunornagnetic beads were provided by Miltenyi Biotec, Germany; mouse-anti-human albumin by Sigma, USA; basic fibroblast growth factor (bFGF) and hepatocyte growth factor (HGF) by PEPROTECH, UK.METHODS: Postembryonic pluripotant stem cells obtained from fetal bone marrow were isolated using density gradient centrifugation and micromagnetic beads technique. The hepatocyte-like cells were induced and differentiated with culture media containing HGF (30 ng/mL) and bFGF (20 ng/mL). Twenty-four SCID mice were randomly divided into experimental group and control group with 12 mice in each group. Hepatic injury models were established with intraperitoneal injection of D-galactosamine On the next day, about 106 CD105(+) cells were perfused into liver in situ in the experimental group, and about 106 CD105(-) cells or isovolumic culture medium were perfused in the control group.MAIN OUTCOME MEASURES: Two, seven days, one and three months after the transplantation of cells, human albumin expression in the liver tissue was detected by immunohistochemistry.RESULTS: The immunocytochemical assay of the cells after micromagnetic beads selection showed that the CD105 expression was slightly positive; the doubling time of the cells in the logarithmic growth period was around 30 hours; after being expanded for 10 population doublings, the cells entered decline period. The cells were transplanted into SCID mice's liver; 3 months later, the human serum albumin in the mouse liver was assessed by using monoclonal antibody of mouse-anti-human serum albumin, dotted or small focal expression of the protein could be detected. However, any expression was not observed in the control group.CONCLUSION: The bone marrow-derived pluripotent stem cells are able to transform to hepatocytas in the hepatic microenvironment.

Objective To investigate clinical characteristics of the multi-center breast DCIS (ductal carcinoma in situ), in order to improve the rate of early diagnosis. Methods Clinical data of 23 multi-center breast DCIS patients admitted and operated on from September 2003 to February 2009 at our department were retrospectively reviewed. Results There were 4 cases associated with nipple discharge,and 18 cases with preoperative negative physical examination (78%). In the preoperative examination within 6 months, the positive rate of mammography was 90% mainly characterized by the existence of diffuse and multiple small calcifications. The positive rate of MRI was 25%, B ultrasound had no positive findings. The patients that were confirmed multi-center breast DCIS by intraoperative frozen section examination received mastectomy and ipsilateral axillary L1, L2 lymph node dissection. Conclusion The diffuse and multiple small calcification is one of early clinical characteristics of multi-center breast DCIS. Although the disease has a wide range of lesion, it still is an early tumor, and has a favourite prognosis after comprehensive treatment.

The effect of pH on the extracellularpolysaccharide synthesis by Aureobasidium pullulans was studied by addding CaCO 3and HCl.Cultivated in P2 liquid medium for 24h,pH dropped to 3.6 because of strongly producing acid.Under this low pH environment,further fermentation for 120h,only 5.9g/L of polysacharide was obtained.When grown in MP2 medium contaning0.5% CaCO 3,the pH was kept above 5.0 during 144 hours,production of polysaccharide increased to 31g/L. The detailed information of effects of controlled pH on polysaccharide production showed an optimal pH value 5.0 must be maintained through the fermentative period.

· AIM: To investigate the efficacy and safety of intravitreal injection of plasmin, hyaluronidase, or the combination of the two in inducing posterior vitreous detachment (PVD).· METHODS: 15 mini-type pigs were assigned to three groups (Group A, B and C), 5 in each group. One eye of each pig was intravitreally injected with the studying agent,and the fellow eye was used as control. Group A received a vitreous injection of hyaluronidase 50U (0.1mL); group B received plasmin 0.5U (0.1mL); group C received plasmin 0.5U (0.05mL) combined with hyaluronidase 50U (0.05mL). The fellow eyes in each group were injected with 0.1mL balanced salt solution (BSS). All the pigs were examined with slit-lamp biomicroscope, direct and indirect ophthalmoscope, B-scan and electroretinograph (ERG). After 7 days, the animals were killed and the eyes were enucleated and examined with light microscope, transmission electron microscope and scanning electron microscope.· RESULTS: B-scan examination showed that one eye of Group A and two eyes of Group B had partial PVD at 1st day after injection and one eye of Group C at 1 hour after injection. On the 7th day, B-scan, light microscopy and scanning electron microscopy demonstrated that all the eyes of Group A and Group B had partial PVD, while none of the control eyes had PVD. Rank sum test for scanning electron microscopy results of all the groups showed P ＜0.005.Furthermore, the comparisons between every two groups were made. The results of analyses were as follows: P＞0.05 between the drug injected eyes of Group A and Group B, P＜0.05 between Group B and Group C, Group A and Group C.The b-wave and a-wave amplitudes of ERG showed no significant difference either between preinjection and postinjection in all groups or between the drug injected eyes and the control eyes in each group. Light microscopy and transmission electron microscopy revealed no damage to the retinal structure.· CONCLUSION: Intravitreal injection of hyaluronidase 50U or plasmin 0.5U or their combination can produce PVD effectively and quickly without retinal functional or structural toxicity. The combination of the two proteases was proved to be synergetic.

Humans ; Hyoid Bone ; surgery ; Sleep Apnea, Obstructive ; surgery

BACKGROUND: Property of artificial liver reactor is generally evaluated by using an enclosed experimental device, which is constructed through simulating bioartificial liver supporting system. Moreover, the enclosed experimental device is also comprehensively evaluated biological function of hepatocytes and biological response efficiency. In addition, it is significance for primarily evaluating the properties of polysulfone membrane hollow fiber reactor to optimize bioartificial liver supporting system. OBJECTIVE: To construct polysulfone membrane bioreactor experimental system, understand the effects on plasma of patients with severe hepatitis, and observe the feasibility of hollow fiber reactor regarded as bioartificial liver reactor. DESIGN: Repeated measurement.SETTING: General Infectious Disease Institute, Southwest Hospital, the Third Military Medical University of Chinese PLA. MATERIALS: Seven Chinese mini-pigs with 1-7 days old and of either gender were provided by Experimental Animal Center, the Third Military Medical University (certification: F99017). The animal disposal accorded to the ethical standard. Tumor necrosis factor-α (TNF-α) and transforming growth factor-β1 (TGF-β1) kits were purchased from Jingmei Bioengineering Company, China; polysulfone membrane hollow fiber reactor by Shanghai Dehong Biomaterial Institute, China; Cellco culture-circular artificial capillary culture system by Spectrum Company, USA; seven plasma exponents were taken from admission patients with chronic severe hepatitis during plasma exchange. All patients provided the informed consent, and the animal experiment was conducted with confirmed consent by the local ethics committee.METHODS: This study was performed at the General Infectious Disease Institute, Southwest Hospital, the Third Military Medical University of Chinese PLA from January 2004 to July 2005. ① Experimental procedure: The experimental pigs underwent ablactation at 12 hours before experiment, and then hepatocytes were separated after cleaning their fur. An enclosed system was composed of polysulfone membrane hollow fiber reactor and Cellco culture-circular artificial capillary culture system, which were connected with oxygen-carbon dioxide diffusion tube and media holding pond. Interface of external cavity was blocked with clean rubber tampon. The separated hepatocytes underwent spheroid culture by using magnetic stirring method, and then they were inoculated in the external cavity of polysulfone membrane hollow fiber reactor. Next, 200 mL plasma from severe hepatitis patients was added in media holding pond and circled from internal cavity of reactor to media holding pond at the speed of 80 mL/min. ② Experimental evaluation: 2 mL liquid circulation separately collected from internal cavity of reactor at 0, 2, 4 and 6 hours after circulation was used to measure level of ammonia in supernatant by using glutamic dehydrogenase-violet method. In addition, content of total bilirubin was detected by using automatic biochemistry analyzer, prothrombin time by using automatic blood coagulometer, and TNF-α and TGF-β1 were detected according to the kits. MAIN OUTCOME MEASURES: Effects of polysulfone membrane bioreactor on ammonia, bilirubin, prothrombin time, TNF-α and TGF-β1 in plasma of patients with severe hepatitis.RESULTS: ① Measurement of ammonia, total bilirubin and prothrombin time in patients with severe hepatitis: Level of ammonia decreased persistently, in particularly, the decrease was obvious from 0 to 2 hours, and then, it was decreased slowly. While, level of total bilirubin was also decreased persistently, and it was significantly lower at 6 hours than that at 0 hour (P < 0.05). Moreover, prothrombin time was decreased persistently, and it was significantly lower at 6 hours than that at 0 hour (P < 0.05). ② Measurement of TNF-α and TGF-β1 in patients with severe hepatitis: Contents of both TNF-α and TGF-β1 decreased persistently, and they were significantly lower at 6 hours than those at 0 hour (P < 0.05). CONCLUSION:Polysulfone membrane bioreactor experimental system can clear noxious substance of small molecules in plasma of patients with severe hepatitis, supply beneficial components, and decrease levels of cytokines. Therefore, it can be regarded as the bioartificial liver reactor.

Objective To investigate the expression of Caspase-9 and heat-shock protein-90 (HSP 90) in rats after focal cerebral ischemia-reperfusion injury.Methods The male SD rats (200-250 g) were divided into three groups by the random number table: normal group, sham group and cerebral ischemia-reperfusion (CIR) group.Each group was sorted into four subgroups including group 6 h, group 24 h, group 48 h and group 72 h according to the reperfusion time.Suture-occluded method was adopted to prepare focal cerebral ischemia-reperfusion(CIR) injury in rat model.Enzymelinked immunosorbent assay (ELISA) method was used to detect the variations of Caspase-9 and HSP-90 expression in rats.Results The changes in Caspase-9 and HSP 90 expression in the brain cells were observed by ELISA method.The expression of Caspase-9 and HSP-90 was weakly expressed in sham group, and was at peak in CIR group within 24 h-48 h, then began to decline at 72 h after the reperfusion time.The differences in the expression of caspase-9 and HSP-70 between sham group and normal group were not statistically significant.Conclusions Apoptotic cells gradually increase along with reperfusion time and reach the peak at 48 h after cerebral ischemia-reperfusion.In ischemia half dark stripe, the expression of Caspase-9 and HSP 90 is increased in neuronal cells after cerebral ischemia-reperfusion, and the positive cells number is at peak at 48 h after cerebral ischemiareperfusion.Apoptosis of neuronal cells after cerebral ischemia and reperfusion is a dynamic evolutionary process.The expression of Caspase-9 and HSP 90 in nerve cells plays an important role in regulating cell apoptosis.

Objective To study the effect of bromocriptine(BRC) on the activation of T lymphocyte stimulated by phytohemagglutinin(PHA).Methods After CD4+ T cell line Jurkat E6-1 cells were stimulated by PHA,prolactin(PRL) and BRC,respectively,the expression of linker for activation of T cells(LAT) and zeta-chain T cell receptor associated protein kinase 70 000(ZAP-70) mRNA of T lymphocytes were checked by RT-PCR.The expression of PRL mRNA of T lymphocytes was detected by Real time PCR.The expression of CD25(cluster of differentiation) as a marker of early activation on the surface of T lymphocytes was detected by flow cytometry,and the activation of nuclear factor-?B(NF-?B) was detected by luciferase reporter system.Results 1.BRC inhibited the expression of ZAP-70 as the common signal molecules both in the T lymphocyte activation pathway and PRL-prolactin-prolactin receptor(PRLR) signal transduction pathway,and decreased the expression of PRL mRNA produced by activation T lymphocytes.2.BRC enhanced the expression of LAT mRNA as another important signal molecular on the T lymphocytes and CD25 on the surface of the T lymphocytes.3.The activation of NF-?B of T lymphocytes was decreased.Conclusions BRC might inhibit the activation of T lymphocytes by inhibiting the expression of ZAP-70,the common signal molecules between T lymphocytes activation and PRL-PRL pathway,and PRL mRNA,the like-T lymphocyte growth factor.

Objective To explore the prognostic value of B-type natriuretic peptide (BNP) for 28-day mortality of elderly patients with non-cardiac critical ill in emergency intensive care unit (EICU). Methods A total of 70 elderly non-cardiac critically ill patients (age≥60 years) in EICU were enrolled, and the blood samples were collected to detect BNP level after the patients' admission to EICU. After 28 days, the mortality was assessed. Results Twenty-two patients (31.4 %) died during 28 days observation, whose BNP levels were significantly higher than that of the survivors [ln BNP: (6.4 ± 1.2) ng/L vs. ( 5. 1 ± 1.5 ) ng/L, P＜ 0. 05] ; BNP level had an area under the receiver operating characteristic curve of 0. 759 (95% CI: 0. 636-0. 882, P＜0.05) for predicting mortality,and the optimal cut point of BNP was 342 ng/L (sensitivity 77.3%, specificity 68.7%).Conclusions BNP level could be a predictor for 28-days mortality for elderly non-cardiac critically ill patients.

ObjectiveTo investigate the expression of programmed death receptor ligand 1 ( PD-L1 ) of peripheral blood mononuclear cells (PBMCs) in patients with hepatic cystic echincccccosis (HCE) and its relation with interferon-γ.MethodsThe clinical data of 63 patients with HCE who were admitted to the First Affiliated Hospital of Xinjiang Medical University from June 2010 to February 2011 were retrospectively analyzed.All patients were divided into HCE active group (38 patients) and HCE non-active group (25 patients) according to the system established by the World Health Organization's Informal Working Group on Echinocoecosis.Twenty patients with hepatic hemangioma or healthy individuals were recruited in normal control group.The positive rate of PD-L1 expression was detected by flow cytometry and immunocytochemistry.The expression of interferon-γ was detected by enzyme-linked immtmosorbent assay (ELISA).All data were analyzed by the t test,one-way analysis of variance,LSD test and chi-square test.The relationship between the expression of interferon-γ and positive rate of PD-L1 expression was analyzed by the Pearson test.ResultsThe results of flow cytometry showed that the positive rates of PD-L1 expression in the HCE active group,HCE non-active group and normal control group were 12.1％±3.8％,10.9％ ± 2.5％ and 9.1％ ±2.5％,respectively.There was a significant difference in the positive rate of PD-L1 expression between the HCE active group and normal control group (t =3.327,P ＜ 0.05 ).The results of immunohistochemistry showed that the positive rates of PD-LI expression in the HCE active group,HCE non-active group and normal control group were 11.9％ ± 3.4％,i0.6％ ± 2.9％ and 9.5％ ± 3.6％,respectively.There was a significant difference in the positive rate of PD-L1 expression between the HCE active group and normal control group (t =2.470,P ＜ 0.05 ).The expressions of intefferon-γ in the HCE active group,HCE non-active group and normal control group were ( 141 ± 38 ) μμg/L,( 124 ± 32 ) μg/L and ( 105 ± 42 ) μg/L.There wasasignificant difference in the expression of interferon-γ between the HCE active group and normal control group ( t =3.280,P ＜ 0.05).The results of flow cytometry and immunohistochemistry revealed that the positive rate of PD-L1 expression was positively correlated with the expression of interferon-γ( r =0.59,0.61,P ＜ 0.05 ).Conclusion With the help of interferon-γ,PD-L1 may play an important role in promoting the immune.evasion of echinococcus.