BACKGROUND: Recently, little attention has been paid to how to induce and identify the functions of differentiated cells in the methods for embryonic stem (ES) cells differentiation into hepatocytes. Whether the differentiated cells express functional characteristics of hepatocytes should be one of the markers to identify the hepatic differentiation of ES cells.OBJECTIVE: To direct mouse embryonic stem cells in vitro differentiation into functional hepatocytes by introduction of murine cholestatic serum in hepatocyte growth factor (HGF)-induced system.DESIGN: A controlled observation and in vitro cytological trial.SETTING: Department of Hepatobiliary Surgery, the Second Affiliated Hospital of Sun Yat-sen University.MATERIALS: The experiment was carried out in the Medical Research Center of the Second Affiliated Hospital of Sun Yat-sen University from October 2004 to February 2007. The mouse E14 ES cell line was kindly provided by the Stem Cell and Tissue Engineering Center of Sun Yat-sen University. Twenty male SD rats, aged 2 weeks, were purchased from the Experimental Animal Center of Sun Yat-sen University. All animal experimental procedures were abided by the rules of animal ethnics.METHODS: The SD rats were undergone common bile duct ligation to induce cholestasis. Ten days after the operation, the whole blood of rats was collected to prepare cholestatic serum. The ES cells were cultured using hanging-drop method for 5-7 days to develop embryonic bodies (EBs). The dissociated EBs cells were then induced hepatic differentiation with spontaneous system, HGF (20 μg/L) system and cholestatic serum (5%) plus HGF (20 μg/L) system, respectively.MAIN OUTCOME MEASURES: The cellular morphologic changes were observed using transverse microscopy dynamically. (2) The cell staining for albumin, α-fetoprotein, CK18/19, glycogen, indocyanine green (ICG) and fluorescein diacetate (FDA) was done after 4 weeks differentiation. (3) The hepatocyte-specific metabolic functions of synthesizing albumin, triacylglycerol and urea nitrogen were assayed at 3 days interval.RESULTS: (1) The differentiation of ES cells cultured in spontaneous system was uncontrolled and the cells could grow into a wide range of three-germ cells. The HGF could promote ES cells differentiation into endoderm and mesoderm (myocardium). But the differentiated cells only expressed low levels of hepatic specific functions in these two induced systems. (2) Under cholestatic serum plus HGF system, the ES cells could differentiate into polygonal cells with very uniform morphology which were positive in glycogen, ICG and FDA staining and showed higher capabilities of synthesizing albumin, triacylglycerol and urea nitrogen than the differentiated cells in the other systems (P＜0.05-0.01).CONCLUSION: The cholestatic serum, a mimic pathological microenvironment in vitro, could effectively promote ES cells-derived hepatocytes induced by HGF to express high level of liver-specific metabolism functions.

Objective To explore the application of CT value in calculating the proton incident energy in proton treatment planning system. Methods Bethe-Block formula and the formula for calculating the proton range were analyzed to study the correlation of the range of proton beam ( 70-250 MeV ) between a variety of radiation equivalent material and water. Procedure of Monte Carlo SRIM2008 was used to verify the possibility of a constant proportional coefficient of range ( Ci ). The proportional coefficient ( Ci ) of range in radiation-equivalent material and the CT value were fitted by using Origin 8.0 software to study the functional relation of CT value and Ci. The actual range of proton was equivalent to a range of water and incident proton energy could be calculated. Results There was a constant range of Ci of proton beam (70-250 MeV) between a variety of radiation equivalent material and water. There was a functional relation between CT value and Ci ( r = 0.999). The actual range of proton in radiation equivalent material can be equivalent to a range of the water. Conclusions CT values and a range of proportional coefficient ( Ci ),and the actual required range of the tumor could be used to accurately calculate the water equivalent range,and the incident proton energy to the position of Bragg peak. A new exploration for using CT technology in proton treatment planning system could be obtained.

Objective To monitor the protective qualities of personal protective appliances and to ensure the health and safety of radiological working personnel.Methods The lead-equivalent thickness of personal protective appliances and materials was measured by means of standard lead slices.The lead equivalent thickness represents in terms of mm Pb.Results 77 pieces of products and samples were measured altogether.The results indicate that the specific lead equivalents of lead-rubber plates were between 0.20-0.39 mm Pb/mm for 37 pieces of lead-rubber plates and the values of 6 pieces of samples were less than 0.25 mm Pb/mm,which did not accord with the requirement of the relational standard.27 pieces of personal protection appliances were measured altogether.They were 12 pieces of protective clothes,4 pieces of protective headgears,5 pieces of protective neckpieces,4 pieces of protective gloves and 2 pieces of protective masks.13 pairs of lead-glass spectacles among them were measured altogether.The measured results for personal protective appliances and lead-glass spectacles showed that actually measured lead-equivalent were higher than the nominal lead-equivalent.Conclusions The protective qualities are reliable for personal protection materials and appliances to be made in home and imported abroad.But the protective qualities of interventional protection gloves should be improved and made them better.

AIM: To investigate whether a pathological micro-environmental culture system consisting of cholestatic sera induces embryonic stem cells (ESC) to differentiate into hepatocyte-like cells in vitro, and select hepatic stem cells from differentiating embryonic stem cells. METHODS: Mouse ESC, E14 cell line, were cultured in Dulbecco's modified Eagle's medium containing 106 U/L recombinant mouse leukemia inhibitory factor (rmLIF) and 10% FCS. After embryonic bodies formed by the hanging drop culture method, they were exposed to fibroblast growth factor-4 (FGF-4) and hepatocyte growth factor (HGF) for one week, and then placed to a pathological micro-environmental culture system consisting of 5% cholestatic sera and cultured for 2 weeks. Morphological examination, immunocytochemical staining of albumin and CK8/18 were carried out, and mRNA level of albumin and transthyretin were detected by RT-PCR. Glycogen storage and urea synthesis of the cells were tested with PAS staining and colorimetric assay, respectively. RESULTS: The proliferation of cells was inhibited at the early stage when cultured in a pathological micro-environmental culture system consisting of 5% cholestatic sera, but 2 weeks later, a large number of epithelial-like cell colonies were observed, which exhibited hepatocellular phenotype, expressing albumin and CK8/18, transcribing mRNA of albumin and transthyretin and synthesizing glycogen and urea. CONCLUSION: A pathological micro- environmental culture system consisting of 5% cholestatic sera could not only induce embryonic stem cells to differentiate into hepatocyte-like cells, but select hepatic stem cells from differentiating embryonic stem cells initially induced by FGF-4 and HGF in vitro as well.

Aims To compare hippocampus between CUMS rats and normal rats, to find differentially ex-pressed proteins and to explore the pathogenesis of de-pression in the protein levels and biological marker. Methods Chronic unpredicted mild stress was taken to establish rat depression model. The ITRAQ-labeled proteins and peptides were separated by the cation col-umn, and differentially expressed proteins were detec-ted and identified by 2D LC-MS/MS. The functions of proteins were analyzed by bioinformatics. Results To-tally 5 109 proteins were identified, 33 differentially expressed proteins were identified, the expressions of 8 proteins were increased and 25 proteins downregulated. Conclusion ITRAQ based sereening is effective in discovering the nosogenesis of depression and new bio-logical marker.

BACKGROUND: The immature dendritic cell (imDC) can induce immunological tolerance and has widely application in the field of organ transplant. At present, the methods of inducing imDC are insufficient, so the new induction method is demanding.OBJECTIVE: To investigate the effect of sodium butyrate (SB) on the maturation and immunological function of human peripheral blood-derived imDC.DESIGN: Controlled observation and in vitro cytological trial.SETTING: Department of Hepatobiliary Surgery in the Second Affiliated Hospital of Sun Yat-sen University.MATERIALS: Five samples of human peripheral blood were obtained from the healthy volunteers (aged 20-23 years) of Sun Yat-sen University, totally 500 mL. Then peripheral blood mononuclear cells (PBMCs) and lymphocytes were isolated within 2 hours.METHODS: The experiment was carried out in the Medical Research Center of the Second Hospital Affiliated to Sun (1 mmol/L) was added for induction, while those supplemented with maturation promoting factor lipopolysaccharide (LPS)the beginning of induction, while LPS was added on the sixth day for second stimulation.MAIN OUTCOME MEASURES: Cell morphological change, flow cytometry was used to detect DC phenotype,FITC-labeled Dextran was used to detect the endocytosis of DC, the production of IL-12 was determined by means of enzyme-linked immunosorbent assay, and the proliferation of lymphocyte induced by DC was assayed with mixed lymphocyte reaction.expressions of CD80, CD83 and HLA-DR were significantly lower in the imDC of routine induction group following SB maturity promoting, compared with LPS group (P＜0.01). On the sixth day, LPS was added into the SB-induced imDC,Endocytosis of DC: The imDC of routine induction group possessed a significantly lower endocytic activity after induced by LPS, and there were extremely significant differences compared with blank control group and SB maturation Production of IL-12: The production of IL-12 in the mDC induced by LPS was significantly higher than that in control group, SB maturation promoting group and SB induction group, the mDC induced by LPS in routine induction group stimulated significantly stronger proliferation of lymphocyte (P＜0.01).

AIM: To investigate the immunological function of sodium butyrate-induced immature dendritic cells in vitro.METHODS: The human monocyte-derived dendritic cells were induced in the presence of human granulocyte macrophage-colony stimulating factor(GM-CSF) and interleukin-4 (IL-4), combined with sodium butyrate. The immunological function of sodium butyrate-induced dendritic cells was detected by the FCM, endocytic activity, T cells stimulatory proliferation capacity, and interleukin-12 (IL-12) production.RESULTS: Sodium butyrate could down-regulate the major histocompatibility complex(MHC) class II and costimulatory molecules of dendritic cells, increase the endocytic activity, induce a stage of T-cell anergey, and inhibit the T helper cell type 1-skewing factor IL-12 production. CONCLUSION: Sodium butyrate inhibits the maturation of dendritic cells and induces production of immature dendritic cells, which may help to explore the machenism of its epigenitic modification.

Objective:To investigate the clinical and genetic characteristics of families with myotonic dystrophy type I.Methods:Two families with myotonic dystrophy type I admitted to Department of Neurology, First Affiliated Hospital of He'nan University of Science and Technology in September and October 2021 were chosen; their clinical data were retrospectively analyzed. The muscle pathological changes were confirmed by electromyography and muscle MRI. Repeat-primed PCR assay was used to detect the number of CTG repeats in the 3' end non-coding region of DMPK gene. Results:Clinical heterogeneity existed among the two families of patients; muscle weakness and muscular atrophy of the skeletal muscle were the main clinical manifestations; limb weakness, axe face, percussion myotonia and myoglobular sign were noted; systemic multi-system symptoms included palpitation and chest tightness, cataracts, gastrointestinal symptoms, fatigue/lethargy, and cognitive impairment. Electromyography showed myotonic potential and myogenic damage. Muscle fatty infiltration and atrophy were noted in muscle MRI, and lesions were predominantly in the gastrocnemius. All patients had abnormal amplification of DMPK gene CTG (number of CTG repeats> 50 or 100). Conclusion:In addition to skeletal muscle involvement, systemic multi-system involvement such as cardiac, eye, respiratory, endocrine, and nervous system should also be noted by clinicians in patients with myotonic dystrophy type I.

Objective:To investigate whether silencing signal transducer and activator of transcription 6 (STAT6) with siRNA can inhibit eosinophilic inflammation of sinonasal mucosa in a mouse model of eosinophilic chronic rhinosinusitis (ECRS).Methods:The study was conducted from March to September in 2022. Forty-eight female BALB/c mice were randomly divided into four groups: the control group, the Vehicle (transfection reagent)-treated group, the Scramble siRNA (Control siRNA)-treated group, and the STAT6 siRNA-treated group, with twelve mice in each group. An ovalbumin (OVA)-staphylococcal enterotoxin B (SEB)-induced ECRS murine model was established. SiRNA prepared in Lipofectamine was locally administered to the nasal cavity. After administration, samples of the peripheral blood and sinonasal mucosa were collected. Eosinophils in peripheral blood were detected by hematology analyzer. Total and OVA-specific IgE (OVA-sIgE) in serum were detected by enzyme-linked immunosorbent assay (ELISA). Mucosal levels of cytokines and chemokines, including interleukin (IL)-5, IL-17A, interferon-γ (IFN-γ) and eotaxin-1, were also measured using ELISA. Mucosal histological changes of eosinophil infiltration were examined using hematoxylin, and eosin staining, and tissue eosinophil count was performed using a microscope under a high-power field (HPF). Tissue expression of STAT6 and phosphorylated STAT6 (p-STAT6) was detected with the western blot method. Immunofluorescence staining was used to localize the expression of p-STAT6 in sinonasal mucosa. Statistical analysis was conducted using SPSS 18.0 software.Results:Peripheral blood eosinophil count, percentage of peripheral blood eosinophil, total serum IgE level, and serum OVA-sIgE level in the STAT6 siRNA-treated group [(0.318±0.045)×10 3/μl, (3.667±0.479)%, (102.070±13.205) ng/ml, and (38.870±7.352) ng/ml] were significantly different from those of the Vehicle-treated group [(0.532±0.049)×10 3/μl, (6.710±1.061)%, (203.102±29.653) ng/ml, and (74.575±6.432) ng/ml, Z value was -2.56, -2.24, -2.40, and -2.56, respectively, all P<0.05] and Scramble siRNA-treated group [(0.493±0.036)×10 3/μl, (5.858±0.872)%, (189.964±30.042) ng/ml, and (80.935±8.358) ng/ml, Z value was -2.17, -2.08, -2.24, and -2.72, respectively, all P<0.05]. Besides, IL-5 and eotaxin-1 levels in the STAT6 siRNA-treated group [(312.279±34.281) pg/ml and (25.297±4.323) pg/ml] were significantly lower than those in the Vehicle-treated group [(689.667±31.905) pg/ml and (68.278±6.485) pg/ml, Z value was -2.73 and -2.88, respectively, both P<0.01] and Scramble siRNA-treated group [(661.783±42.094) pg/ml and (63.015±7.416) pg/ml, Z value was -2.72 and -2.81, respectively, both P<0.01]. Tissue eosinophil count in sinonasal mucosa was (29.132±4.163)/HPF in the STAT6 siRNA-treated group, and were significantly less than those in the Vehicle-treated group [(78.050±7.912)/HPF, Z=-2.88, P<0.01] and Scramble siRNA-treated group [(73.864±8.671)/HPF, Z=-2.72, P<0.01]. The expression level of STAT6 protein (0.105±0.021) was significantly decreased in the mice treated with STAT6 siRNA compared with PBS, Vehicle, and Scramble siRNA (0.232±0.037, 0.243±0.039, and 0.228±0.032, Z value was -2.25, -2.49, and -2.56, respectively, all P<0.05). Corresponding, p-STAT6 protein level (0.292±0.038) was markedly decreased by the introduction of STAT6 siRNA, the difference was statistically significant as compared with the Vehicle-and Scramble siRNA-treated groups (0.613±0.046 and 0.641±0.050, Z value was -2.81 and -2.88, respectively, both P<0.01). Immunofluorescence staining showed that p-STAT6 was mainly located in the nucleus of nasal epithelial cells and inflammatory cells. The green fluorescence of p-STAT6 expression in sinonasal mucosa in the STAT6 siRNA-treated group was weaker than that in the Vehicle-and Scramble siRNA-treated groups. Conclusion:Intranasal administration of STAT6 siRNA can significantly downregulate STAT6 expression, decrease p-STAT6 level, and prohibit the development of Th2-skewed ECRS.

OBJECTIVE： To evaluate the effectiveness of trolamine for preventing and treating radiation dermatitis （RD） and evidence quality， and to provide reference for clinical use. METHODS： Retrieved from PubMed， Cochrane library， Embase， CNKI， Wanfang and VIP database， randomized controlled trials （RCTs） about trolamine （trial group） versus usual care （control group） for preventing and treating RD were collected. After data extraction， Cochrane bias risk assessment tool 5.0.2 was used to assess the bias risk， and Rev Man 5.3 statistical software was used to perform the Meta-analysis. GRADE evidence quality grading system was used to evaluate the evidence quality of outcome indexes. RESULTS： Seven RCTs were included， involving 782 patients. Results of Meta-analysis showed that there was no statistical significance in total incidence of RD [OR＝0.50， 95％CI （0.23， 1.11）， P＝0.09]， and the incidence of grade Ⅰ RD [OR＝1.32， 95％CI（0.96，1.81）， P＝0.09]， grade Ⅱ RD [OR＝1.07， 95％CI（0.80，1.42）， P＝0.66]， grade Ⅲ RD [OR＝0.69， 95％CI（0.45，1.04）， P＝0.07] or grade Ⅳ RD [OR＝0.43， 95％CI（0.17，1.05）， P＝0.07] between 2 groups. Results of Grade evidence quality evaluation showed that total incidence of RD， and the incidence of grade Ⅱ RD and grade Ⅳ RD were recommended by moderate-level evidence in 2 groups， while the incidence of grade Ⅰ and grade Ⅲ RD were recommended by low-level evidence. CONCLUSIONS： Trolamine is not effective in preventing and treating RD， and can not reduce the incidence of RD.