To establish a method of directional differentiation and efficient production of neurons from embryonic stem cells (ES cells) in vitro, based on the 4-/4+ protocol described by Bain, a new method was established to induce ES cells differentiating into neurons by means of three-step differentiation using all-trans retinoic acid (ATRA) combined with astrocyte-conditioned medium (ACM) in Vitro. The totipotency of ES cells was identified by observation of cells' morphology and formations of teratoma in immunocompromised mice. The cells' differentiation was evaluated continuously by the detection of the specific cellular markers of neural stem cells, neurons and astrocytes, including nestin, NSE and GFAP using immunohistochemistry assay. The NSE positive cells' ratio of the differentiated cells was determined by flow cytometry. It was found that the transparent circular clusters surrounding embryoid bodies induced with combining induction protocol formed just after 24 h and gradually enlarged later. This phenomenon could not be observed in EBs induced only by ATRA. The NSE positive cells' ratio in the cells induced with ATRA and ACM was higher than that of the cells induced by ATRA at different time points of differentiation, and finally reached up to 73.5% among the total differentiated population. It was concluded that ES cells could be induced into neurons with high purity and yield by means of inducing method combining with ATRA and ACM.
Astrocytes/*cytology ; Cell Differentiation ; Cells, Cultured ; Embryo, Mammalian ; Neurons/*cytology ; Stem Cells/*cytology ; Tretinoin/pharmacology

Background Interleukin-17 (IL-17)is a potent pro-inflammatory cytokine and plays a pathogenic role in autoimmune disease.It was confirmed that IL-17 is implicated in allograft rejection of many transplanted organs.Recent studies have foensed on the effect of IL-17 antagonists on allograft rejection.Objective This study aimed to investigate the inhibitory effect of anti-mouse IL-17 monoclonal antibody (mAb) on corneal allograft rejection.Methods Twenty-five 8 to 10-week-old C57BL/6 mice and 50 BALB/c mice were collected.Donor cornea grafts with 2 mm diameter from 25 C57BL/6 mice was transplanted to 50 eye of BALB/c mice to establish a model of corneal transplantation.The recipients were randomized into 2 groups,and neutralizing mouse IL-17antibody or isotype control antibody was intraperitoneally injected immediately after transplantation for experimental treatment,respectively.Allografts were scored clinically at appropriate time points after treatment based on Plskova criteria,and ≥5 was confirmed as rejection.Infiltrating cells in corneal graft were detected qualitatively and quantitatively by immunohistochemistry and reverse transcription-PCR separately.The cytokine levels of T helper type 1 (Th1),Th2,and Th17 in recipients' spleen wer(c) analyzcd by ELISA.The use of the animals followed the Statement of ARVO.Results Compared with the isotype control antibody group,the survival of grafts was improved in the IL-17mAb group(P＜0.05).The levels of neutrophile granulocyte mRNA,CD4+ and CD8+ T lymphotes mRNA were 2.22±0.10,1.64±0.04 and 1.32±0.10 in the IL-17 mAb group,showing a significant decline in comparison with those of the isotype control antibody group(3.61 ±0.08,2.69±0.06 and 2.17±0.04) (P=0.000,0.000,0.000).Interferon-γ(IFN-γ),IL-12 p40 and IL-17 concentrations in recipients ' splenocytes were (529.80 ± 13.83) ng/L,(539.58 ±10.74) ng/L and(173.70±8.11)ng/L in the IL-17 mAb group,and thosc in the isotype control antibody group were (741.48± 10.51) ng/L,(1156.90 ± 69.93) ng/L and (366.13± 7.93) ng/L,with significant differences between them (P=0.000,0.001,0.000).Conclusions Neutralization IL-17 bioactivity inhibits mouse corneal allograft rejection to a certain extent.

Objective To investigate the prevalence of clonorchiasis among residents of Huachuan Country,Heilongjiang Province and to provide a basis for development of control strategies.Methods From 2011 to 2012,cluster random sampling was performed to survey the incidence of clonorchiasis in Huachuan Country.Fecal specimens were collected and examined the clonorchis sinensis eggs by Kato-Katz method.A questionnaire survey was conducted to collect related information such as age,gender,occupation and eating habits.The infection characteristic was analyzed.Results Totally 884 patients with clonorchiasis were found among 2248 residents,and the infection rate was 39.32％.The infection rate in male[47.15％(611/1296)] was significantly higher than females [28.68％(273/952),x2 =34.55,P ＜ 0.01].The infection rate increased with age,which was higher in the 20-69 years old people,with the highest infection rate in the 50-59 years old groups[45.34％ (219/483)].Of the occupational distribution,farmers had the highest infection rate [47.24％ (420/889)],followed by cadres and staffs[38.38％(190/495)].Of residents with fresh fish eating history,the prevalence of clonorchiasis was 53.38％(150/281).Conclusions The prevalence of clonorchiasis is still high in Huachuan County.To reduce the prevalence of clonorchiasis,comprehensive prevention measures,health education and group chemotherapy should be carried out.

Objective To determine whether homocysteine(Hcy)and monosodium glutamate (MSG)could lead to animal model of hypertensive disorder complicating pregnancy and its mechanism. Methods Female adult Wistar rats were randomly divided into 4 groups:pregnant control group(PN), pregnant Hcy group(PH),pregnant glutamic acid group(PG)and pregnant Hcy and glutamie acid group (PHG).The rats of each group were injected with Hey 200 mg/kg or physiological saline every day intraperitoneally and with MSG or 0.9% saline every other day via Hcy injection from the 10th day to the 20th day of pregnancy.The blood pressure,urine protein,function of liver and kidney,weight of placenta, length and weight of fetus were all measured.The histological change of the pallium and the change of behavior of pregnant rats were also observed.Results(1)The blood pressure in PH[(107?8)mm Hg, 1 mm Hg=0.133 kPa],and PHG group [(109?10)mm Hg] after the treatment increased significantly compared with those in other groups from the 12 th day after pregnancy(P

Saccharmyces cerevisiae 220 has certain resistance to Pb2+ was found through the test of it's cultured on media containing lead cation. Sacchannyces cerevisiae 220 cultured was put into the media cotaining lead cation and it's biosorp-tion to lead cation was studied. The results show this strain has maximum biosorption ratio (96.6%) to 6mg/L lead cation, biosorption-equilibrilium period is 25-30min, biosorption kinitics equation is q-26.318C/ (1 + 0.437C) . Acetic acid is benefit to de-biosorption, glucose and KH2PO4 can improve biosorption ratio to lead cation.

A.niger M1, the initial strain, was treated by UV and a mutant with 30% higher xylanase activity was obtained. Zymogram for detecting xylanase showed there are three different xylanases in the mutant mature culture, while two xylanases in initial strain. After orthogonal experiment, the optimum fermentation conditions of the mutant were obtained as follows: concentration of the major carbon resource 4 %, ratio between bran and corncob 5:5, concentration of glucose 0.1%, concentration of ammonium oxalate as supplemental nitrogen resource 2.0%, the initial pH of liquid medium 5.0, 100mL/250mL flask.

Aged ; Fatal Outcome ; Heart Rupture ; etiology ; Humans ; Male ; Myocardial Infarction ; therapy

OBJECTIVETo investigate clinicopathological and genetic characteristics of primary ocular adnexal lymphoproliferative lesions.

METHODSClinical, morphological and immunohistochemical features of 37 archival cases of primary ocular adnexal lymphoproliferative lesions were studied including 5 cases of reactive lymphoid hyperplasia and 32 lymphomas retrospectively. Classification of the lymphomas were made according to the WHO classification of tumors of haematopoietic and lymphoid tissues. All cases were studied by interphase fluorescence in situ hybridization (FISH) using dual color break apart probes of IgH, MALT1, bcl-6, c-Myc, bcl-2, CCND1, bcl-10, and FOXP1 for detection of chromosomal aberrations involving IgH, MALT1, bcl-6, c-Myc, bcl-2, cyclinD1, bcl-10 and FOXP1 genes, respectively. FISH with IgH / bcl-2 dual color dual fusion probe was used for detection of t(14;18)(q32;q21)/IgH-bcl-2. CEP18 spectrum orange probe was used for detection of aneuploidy of the chromosome 18.

RESULTSAmong 32 cases of lymphomas, 28 cases (87.5%) were extranodal marginal zone B-cell lymphomas of mucosa associated lymphoid tissue (MALT lymphoma), 2 cases were follicular lymphoma (FL) and 2 cases diffuse large B cell lymphoma (DLBCL). Among the 28 cases of MALT lymphoma, chromosomal aberrations were found in 60.7% (17/28) by interphase FISH analysis. One case showed positive IgH break-apart signal with unknown partner. 16 cases showed three copies of different genes, of which, three copies of MALT1, bcl-6, and c-Myc were identified in 7 cases (25%), 12 cases (43%), and 2 cases (8%) of MALT lymphomas, respectively. In addition, 5 cases showed two genes including three copies of bcl-6 and MALT1 in 4 cases, and three copies of bcl-6 together with c-Myc in one case. Furthermore, all cases with three copies of MALT1 had trisomy 18. t(14;18)(q32;q21) was detected in both follicular lymphomas. Of the 2 DLBCL cases, one showed three copies of bcl-6 together with trisomy 18 and the other one showed three copies of bcl-6 together with IgH and c-Myc rearrangements. Chromosomal aberration was not found in all 5 cases of reactive lymphoid hyperplasia.

CONCLUSIONSThe most common entity of primary ocular adnexal lymphomas is MALT lymphoma and FISH is helpful for their differential diagnosis and classification. Trisomy 18 and three copies of bcl-6 are common chromosomal aberrations in primary ocular adnexal MALT lymphomas.

Aneuploidy ; B-Lymphocytes ; pathology ; Caspases ; genetics ; Chromosome Aberrations ; Chromosomes, Human, Pair 14 ; Chromosomes, Human, Pair 18 ; Eye ; pathology ; Eye Neoplasms ; genetics ; pathology ; Female ; Genes, bcl-2 ; genetics ; Humans ; Immunoglobulin Heavy Chains ; genetics ; In Situ Hybridization, Fluorescence ; Interphase ; Lymphoma, B-Cell ; genetics ; Lymphoma, B-Cell, Marginal Zone ; genetics ; Lymphoma, Large B-Cell, Diffuse ; genetics ; pathology ; Male ; Mutation ; Translocation, Genetic ; Trisomy

Background Corneal epithelial abrasion results in corneal ulcer and stroma cloudy evenb irreversible visual impairment.Previous drugs for corneal epithelial injury can only alleviate the inflammatory irritation.So it is very important to seek a drug which regulate the growth of corneal epithelium.Objective This study was to investigate the effects of recombinant human BIGH3 protein eye drops on corneal epithelial abrasion.Methods Fifty right eyes of 50 clean adult New Zealand white rabbits were collected.Two rabbits were sacrificed right away following establishment of corneal epithelial abrasion models (0 hour group).The other 48 rabbits were randomly divided into recombinant human epidermal growth factor (EGF) derivative group (positive control group),normal saline solution group (negative control group),0.25％ or 0.5％ recombinant human BIGH3 protein eye drops group.Corneal abrasion models were created with alcohol corrosion method with a defect area of 7 mm2.The corresponding eye drops were used separately in 4 groups for four times per day after operation.Experimental eyes were examined by the slit lamp microscope,and fluorescein vital staining were performed 12,24,36,48,72 hours after operation.Planimetry was performed and the corneal photographs were analyzed with computer software.The rabbits were sacrificed 12,24,36,48 and 72 hours after operation,respectively,and the histopathological examination of corneal tissue was carried out.Results No obvious irritation response was seen after administered of eye drops in the recombinant human EGF derivative group,normal saline solution group,0.25％ and 0.5％ recombinant human BIGH3 protein eye drops groups.Histopathological examination revealed a full-thickness defect of corneal epithelium after modeling.The defect area was gradually smaller with time lapse,and corneal epithelium migrated from periphery toward the center zone.Corneal epithelial cells increased with time lapse.Compared with normal saline solution group,the defect area of corneal epithelium lessened 12,24,36,48 hours after operation in the 0.25％,0.5％ recombinant human BIGH3 protein eye drops groups and recombinant human EGF derivative group (all at P =0.000),but at 12and 24,36 hours after operation,no significant differences were found between the recombinant human EGF derivative group and normal saline solution group (P =0.321,0.057,0.126).The defect area was smaller in the 0.5％recombinant human BIGH3 protein eye drops group than that of the recombinant human EGF derivative group at various time points (P=0.042,0.039,0.025,0.008).However,significant smaller defect area was exhibited only at 12 hours and 24 hours after operation in the 0.25％ recombinant human BIGH3 protein eye drops group (P=0.047,0.042).No significant differences were seen in corneal defect area at various time points between 0.25％ and 0.5％recombinant human BIGH3 protein eye drops groups (P =0.358,0.259,0.108,0.062).In addition,the corneal defect area was (0.51 ±0.42)mm2 72 hours after operation in the normal saline group;while that in the recombinant human EGF derivative group and recombinant human BIGH3 protein eye drops groups was disappeared.The repairing curves in the recombinant human BIGH3 protein eye drops groups were superior to those of the recombinant human EGF derivative group and normal saline solution group.Conclusions 0.25％ and 0.5％ recombinant human BIGH3 protein eye drops have facilitation effect on the growth of corneal epithelial cells and the healing of corneal injury.

ThisstudywasaimedtoobservetheinhibitoryeffectsofQi-Zhu (QZ)granulesonearlyproteinuria in diabetic nephropathy rats with syndrome of qi-yin deficiency and phlegm blocking collaterals. A total of 44 rats were randomly divided into the blank group, model group, Huang-Kui capsule group, QZ granules group. The rat model of diabetic nephropathy with syndrome of qi-yin deficiency and phlegm blocking collaterals was induced by the combination of unilateral renal artery ligation, diet of high-calorie and high cholesterol, and intraperitoneal injection of low-dose streptozotocin. The medication was given for 8 weeks. The concentrations of protein and creatinine in urine were observed on the 4th week. The blood glucose, blood lipids, liver function and renal pathological changes were observed at the end of the experiment. The results showed that compared with the model group, QZ granules can obvious suppress early proteinuria in diabetic nephropathy, promote creatinine excretion, regulate blood lipid metabolism, protect liver function and improve renal pathological changes. It was concluded that QZ granules had independent inhibition effect on early proteinuria in diabetic nephropathy rats with syndrome of qi-yin deficiency and phlegm blocking collaterals. The effect was independent of lowing blood glucose. It represented the corresponding relation between the syndrome and efficacy in Chinese herb compounds.