Discovery of New Isotopes

Using the fusion-evaporation reaction ^{106}Cd(^{58}Ni,4n)^{160}Os and the gas-filled recoil separator SHANS, two new isotopes _{76}^{160}Os and _{74}^{156}W have been identified. The α decay of ^{160}Os, measured with an α-particle energy of 7080(26) keV and a half-life of 201_{-37}^{+58} µs, is assigned to originate from the ground state. The daughter nucleus ^{156}W is a ß^{+} emitter with a half-life of 291_{-61}^{+86} ms. The newly measured α-decay data allow us to derive α-decay reduced widths (δ^{2}) for the N=84 isotones up to osmium (Z=76), which are found to decrease with increasing atomic number above Z=68. The reduction of δ^{2} is interpreted as evidence for the strengthening of the N=82 shell closure toward the proton drip line, supported by the increase of the neutron-shell gaps predicted in theoretical models.

New α-Emitting Isotope

A new α-emitting isotope ^{214}U, produced by the fusion-evaporation reaction ^{182}W(^{36}Ar,4n)^{214}U, was identified by employing the gas-filled recoil separator SHANS and the recoil-α correlation technique. More precise α-decay properties of even-even nuclei ^{216,218}U were also measured in the reactions of ^{40}Ar, ^{40}Ca beams with ^{180,182,184}W targets. By combining the experimental data, improved α-decay reduced widths δ^{2} for the even-even Po-Pu nuclei in the vicinity of the magic neutron number N=126 are deduced. Their systematic trends are discussed in terms of the N_{p}N_{n} scheme in order to study the influence of proton-neutron interaction on α decay in this region of nuclei. It is strikingly found that the reduced widths of ^{214,216}U are significantly enhanced by a factor of two as compared with the N_{p}N_{n} systematics for the 84≤Z≤90 and N<126 even-even nuclei. The abnormal enhancement is interpreted by the strong monopole interaction between the valence protons and neutrons occupying the π1f_{7/2} and ν1f_{5/2} spin-orbit partner orbits, which is supported by the large-scale shell model calculation.

Clinicopathologic features and outcome of cervical cancer: implications for treatment.

OBJECTIVE: We used a regression analysis of the SEER database to establish a new Nomogram for predicting prognosis of cervical cancer patients and guiding the treatment. PATIENTS AND METHODS: We divided the data into the training cohort and the verification cohort. Univariate and multivariate Cox risk regression analysis was used to identify independent prognostic factors and establish a Nomogram model. The verification cohort was used for external verification, and the accuracy was evaluated with C-index and AUC. Finally, Nomogram was used to establish 1-year, 3-year and 5-year survival curves of cervical cancer patients. RESULTS: In this study, 5691 patients with cervical squamous cell carcinoma were included. Data obtained from the training cohort were independent risk factors of cervical cancer AJCC stage (p = 0.039), RX Summ - Surgery Primary Site (p = 0.012), radiation (p = 0.031), chemotherapy (p = 0.013), tumor size (p = 0.009), race (p = 0.039). The 1-year, 3-year, and 5-year overall survival rates for cervical cancer patients were 77.2%, 47.8%, and 35.2%, respectively. CONCLUSIONS: The Nomogram model can better screen out more reasonable comprehensive treatments for patients at different stages. And it is of great help to improve the survival rate and reduce the recurrence rate of cervical cancer patients.

Short-Lived α-Emitting Isotope

A new, very short-lived neutron-deficient isotope ^{222}Np was produced in the complete-fusion reaction ^{187}Re(^{40}Ar,5n)^{222}Np, and observed at the gas-filled recoil separator SHANS. The new isotope ^{222}Np was identified by employing a recoil-α correlation measurement, and six α-decay chains were established for it. The decay properties of ^{222}Np with E_{α}=10016(33) keV and T_{1/2}=380_{-110}^{+260} ns were determined experimentally. The α-decay systematics of Np isotopes is improved by adding the new data for ^{222}Np, which validates the N=126 shell effect in Np isotopes. The evolution of the N=126 shell closure is discussed in the neutron-deficient nuclei up to Np within the framework of α-decay reduced width.

New Isotope

A new short-lived neutron-deficient isotope ^{220}Np was synthesized in the fusion-evaporation reaction ^{185}Re(^{40}Ar,5n)^{220}Np at the gas-filled recoil separator SHANS. Based on the measurement of the correlated α-decay chains, the decay properties of ^{220}Np with E_{α}=10040(18) keV and T_{1/2}=25_{-7}^{+14} µs were determined, which are in good agreement with theoretical predictions. From the new experimental results coupled with the recently reported α-decay data of ^{219,223}Np, the α-decay systematics for Np isotopes around N=126 was established, which allows us for the first time to test the robustness of the N=126 shell closure in Z=93 Np isotopes. The results also indicate that, in the region of nuclei with Z≥83, the proton drip line has been reached for all odd-Z isotopes up to Np.

[Feasibility and safety of MRgFUS ablation for uterine fibroids and adenomyosis: a preliminary study].

Objective: To investigate the feasibility and safety of magnetic resonance guided focused ultrasound surgery (MRgFUS) ablation treatment for uterine fibroids and adenomyosis. Methods: From February 2017 to July 2018, a total of 61 women in Foshan Hospital of Traditional Chinese Medicine with uterine fibroids or adenomyosis (36 cases of fibroids and 25 cases of adenomyosis) were included for treatment of MRgFUS, mean age was 27-48 (39±5) years. The treatment status, treatment effect and complications were recorded, and the differences between myoma and adenomyosis were compared. Results: Thirty-two (88.9%) patients of fibroid group and twenty-one (84.0%) patients of adenomyosis group were completed MRgFUS treatment respectively (P>0.05). The spot energy of adenomyosis group was 1 039-5 698(2 852±991) J, which was higher than 600-6 466(2 485±1 137) J of fibroid group (P<0.01). There was no significant statistical difference in mean temperature of spot and ablation time between the two groups (P>0.05).The non-perfusion volume ratio (NPVR) of the fibroid and adenomyosis group was 54%-99%(84%±15%) and 60%-98%(82%±12%) and there was no significant statistical difference (P>0.05), but the ablation efficiencies of adenomyosis group was less than fibroid group (0.8-4.3(2.1±0.9) cm(3)/min vs 1.3-7.8(3.6±1.5)cm(3)/min, P<0.01).The incidence of complications of adenomyosis group was 47.6%(10/21), it was higher than fibroid group 18.8%(6/32) (P<0.05). Conclusions: MRgFUS is a non-invasive, safe and effective treatment for both uterine fibroids and adenomyosis. Compared with uterine fibroids, MRgFUS treatment of adenomyosis has some disadvantages such as higher energy, lower ablation efficiency and more adverse reactions, and further optimization is needed.

CT and MRI studies of hepatic angiosarcoma.

AIM: To analyse the computed tomography (CT) and magnetic resonance imaging (MRI) manifestations of hepatic angiosarcoma. MATERIALS AND METHODS: Nineteen patients with hepatic angiosarcoma underwent preoperative abdominal unenhanced and contrast-enhanced CT (11 cases) or (eight cases) MRI. RESULTS: The results of a coagulation examination showed varying degrees of abnormalities in 12 (63.16%) cases (most were prolonged prothrombin time and an increased proportion of prothrombin time), which were the most common abnormalities on the laboratory tests. Unenhanced CT of the lesions showed homogeneous or heterogeneous hypointense with hyperintense haemorrhagic lesions, contrast-enhanced CT was performed for six cases (6/11, 54.5%) with centripetal heterogeneous filling. All lesions showed heterogeneous intensity at MRI, including heterogeneous hypointense T1WI and homogeneous or heterogeneous hyperintense T2WI. Haemorrhage lesions showed higher hyperintensity with spot or patchy signals. Centripetal enhancement was found in six cases using contrast-enhanced imaging. Flaky patches of contrast enhancement were seen in the lesions. CONCLUSION: The CT and MRI features of most of the hepatic angiosarcomas in the present study were relatively characteristic: the border of the mass was indistinct, the density was heterogeneous, and haemorrhage was frequently seen, with secondary calcification in a few cases, whereas enhanced imaging showed typical centripetal heterogeneous enhancement. In addition, highly malignant angiosarcoma could not be enhanced.

FHL2 participates in renal interstitial fibrosis by altering the phenotype of renal tubular epithelial cells via regulating the ß-catenin pathway.

OBJECTIVE: To investigate the potential role of FHL2 (four and a half LIM domains protein 2) in the renal interstitial fibrosis and its underlying mechanism. MATERIALS AND METHODS: NRK-52E, the rat tubular epithelial cell line, was selected for the in vitro experiments. The unilateral ureteral obstruction (UUO) mouse model and phenotype changes of NRK-52E cells were induced by TGF-ß (transforming growth factor-ß) treatment. Protein and mRNA expressions of biomarkers of tubular cells and renal fibrosis in NRK-52E cells were detected. Meanwhile, phenotype changes of NRK-52E cells were detected after FHL2 overexpression. Furthermore, CD1 mice were selected for constructing the UUO mouse model. Protein and mRNA expressions of biomarkers of tubular cells and renal fibrosis in kidney tissues were detected. CD1 mice with FHL2 overexpression were constructed by tail vein injection of FHL2 plasmid for further observation of renal interstitial fibrosis. Expressions of ß-catenin pathway-related genes were detected by Western blot and polymerase chain reaction (PCR), respectively. RESULTS: The FHL2 expression was increased during the phenotype change of NRK-52E cells induced by TGF-ß treatment. Overexpression of FHL2 led to a significant phenotype change. Similarly, the FHL2 expression was elevated in the UUO mouse model. Renal interstitial fibrosis was exaggerated and expression levels of genes related to the ß-catenin pathway were increased after injection of FHL2 plasmid. CONCLUSIONS: FHL2 is involved in renal interstitial fibrosis by altering the phenotype of renal tubular epithelial cells via regulating the ß-catenin pathway.

Silencing of LncRNA TCONS_00088786 reduces renal fibrosis through miR-132.

OBJECTIVE: To examine the role of long non-coding ribonucleic acid (LncRNA) TCONS_00088786 in the development of renal interstitial fibrosis and its potential mechanism in this process. MATERIALS AND METHODS: Unilateral ureteral obstruction (UUO) was used to induce tubulointerstitial fibrosis. Masson staining showed the degree of renal fibrosis in UUO mice. Immunohistochemistry and immunofluorescence were performed to detect the fibrosis-related proteins, the 24-h urine volume and protein content. The renal functions were reflected via serum creatinine (Scr) and blood urea nitrogen (BUN). Changes in lncRNATCONS_00088786, miR-132 and collagen I and III in the development process of renal fibrosis were detected through reverse transcription-polymerase chain reaction (RT-PCR). Small interfering RNA (siRNA) was transfected into NRK52E cells to mimic the knockdown. Western blot was adopted to detect the changes in miR-132, collagen I and III after the siRNA was transfected by transforming growth factor-ß (TGF-ß) for 24 h. RESULTS: With the development of renal fibrosis, lncRNA TCONS_00088786 and miR-132 were increased gradually. After the knockdown of lncRNA TCONS_00088786, miR-132 was decreased and fibrosis-related protein was also decreased. CONCLUSIONS: Decreased lncRNA TCONS _00088786 inhibits renal interstitial fibrosis by reducing miR-132 and it may be a potential novel molecular target for the treatment of renal interstitial fibrosis.

Ablation of carbohydrate-responsive element-binding protein improves kidney injury in streptozotocin-induced diabetic mice.

OBJECTIVE: Carbohydrate-responsive element-binding protein (ChREBP) has been reported to regulate glucose and lipids metabolism in the liver. However, its role in the complicated pathophysiology of diabetic nephropathy is not understood. MATERIALS AND METHODS: C57BL/6 mice treated with streptozotocin (STZ) or vehicle control to induce diabetic models. The mRNA and protein levels of ChREBP in kidneys of control and diabetic mice were determined by real-time PCR or Western Blot, respectively. The expression of inflammatory and endoplasmic reticulum stress markers in ChREBP deficient or Wild-type mice was also determined by real-time PCR or Western Blot. Urine was collected over 16 hours on the day prior to sacrifice, and albuminuria and urine creatinine were determined by Elisa or Creatinine Assay Kit. RESULTS: We found that expression of ChREBP and its downstream target genes were up-regulated in C57BL/6 mice with diabetic nephropathy. Subsequently, we demonstrated that ChREBP knockout mice were protected against the development of diabetic nephropathy induced by streptozotocin (STZ), showing less albuminuria, inflammation and glomerular hypertrophy as compared to diabetic wild-type mice. Reduced expression of inflammatory and endoplasmic reticulum stress markers were also observed in ChREBP deficient mice. CONCLUSIONS: Our data indicate that ablation or inhibition of ChREBP might improve kidney injury in streptozotocin-induced diabetic animals.

Evidence for Octupole Correlations in Multiple Chiral Doublet Bands.

Two pairs of positive-and negative-parity doublet bands together with eight strong electric dipole transitions linking their yrast positive- and negative-parity bands have been identified in ^{78}Br. They are interpreted as multiple chiral doublet bands with octupole correlations, which is supported by the microscopic multidimensionally-constrained covariant density functional theory and triaxial particle rotor model calculations. This observation reports the first example of chiral geometry in octupole soft nuclei.

[Clinico-pathological characteristics and prognosis of IgA nephropathy patients with microalbuminuria and deposition of complement C3].

OBJECTIVE: To analyze the clinical and pathological data and prognosis of IgA nephropathy patients with microalbuminuria and deposition of C3, and to investigate the significance of C3 deposition in IgA nephropathy with microalbuminuria. METHODS: The clinical and pathological data of 127 IgA nephropathy patients with microalbuminuria confirmed by renal biopsy in the Jining No.1 People's Hospital from January 2009 to January 2015 and minimum 6-month follow-up was reviewed, and patients were divided into positive group (72 cases, 56.7%)and negative group (55 cases, 43.3%) according to the deposition of C3 in the mesangial area of glomeruli. 24 h urine quantitative protein being more than 1 g, or serum creatinine level becoming abnormal or double by renal biopsy was defined as endpoint of follow-up. Renal survival was calculated by Kaplan-Meier survival analysis. RESULTS: A total of 127 IgA nephropathy patients with microalbuminuria were followed up successfully, with an average follow-up of (49.6±22.7) months. 24 h urine albumin[(261.3±47.4) vs (238.7±51.9) mg, P=0.011], serum creatinine value[98.0(56.4, 118.6) vs 85.7(51.9, 107.8) µmol/L, P=0.003], uric acid value[(384.0±93.7) vs (360.5±88.4) µmol/L, P=0.043] and serum IgA value[(3.36±1.17) vs (3.12±1.05) g/L, P=0.044] were significantly higher in the C3 positive group than those in the negative group, while the serum complement C3 was significantly lower [(0.70±0.42) vs (0.98±0.49) mg, P=0.047]. Pathological changes [Lee's grade Ⅲ and above Ⅲ: 21(16.5%) vs 11(8.7%), P=0.034], glomerular sclerosis or adhesions [29(22.8%) vs 19(15.0%), P=0.047], renal tubular atrophy or interstitial fibrosis [13(10.2%) vs 8(6.3%), P=0.027] and crescent formation [7(5.5%) vs 2(1.6%), P=0.035] in the complement C3 positive group were more severe than those in the negative group. 38 cases of complement C3 positive group and 14 cases of negative group accomplished the study, and Kaplan-Meier survival analysis showed that there was a significant difference in the median survival time between the two groups [(52.6±8.9) vs (66.1±9.7) months, P=0.019]. CONCLUSIONS: The clinical and pathological features in IgA nephropathy patients with microalbuminuria and deposition of complement C3 were more severe than those without complement C3 deposition, and the prognosis was not optimistic. Therefore, early and active intervention treatment should be considered for these patients.

[Curative effect analysis of posterior decompression and internal fixation for spinal metastases epidural spinal cord compression].

OBJECTIVE: To explore the effects of posterior decompression and internal fixation for spinal metastases epidural spinal cord compression (MESCC) and analyze the related factors of postoperative ambulation function. METHODS: Clinical data of 67 cases with MESCC who received thoracic posterior decompression and internal fixation in our department from January 2006 to December 2014 was retrospectively analyzed. Information about patients' age, gender, pathological type of primary tumor, Karnofsky performance status (KPS) score, pre-operative and postoperative visual analogue scale, preoperative Frankel grade, pre-operative and postoperative imaging characteristics (number of thoracic vertebrae metastases, location, compression fractures of vertebral bodies), time of movement dysfunction and survival was collected. RESULTS: At the end of the follow-up of 67 cases, 57 cases were dead, 10 cases were alive, and the median survival was 8.1 months (1.2-91.9 months).38 cases (67%) died within one year, 50 cases (88%) died within two years. Visual analogue scale of preoperative and postoperative dropped from (5.67±1.67) points to (2.11±1.39) points (P<0.001), 38 (53%) patients' Frankel grade improved at least one grade. Among the 34 cases who were unable to walk, 15 cases regained ability of ambulation after surgery. The patients with KPS scores greater than 80 points and/or had preoperative ambulation ability, tended to have better postoperative ambulatory function. CONCLUSIONS: Posterior decompression and internal fixation for MESCC is effective, and can effectively relieve pain and spinal cord compression, improve neurological function and the quality of life. The ambulatory functional outcomes after surgery are dependent on KPS scores, the occurrence time of neurological dysfunction, preoperative ambulatory status.

[Comparison of curative effect and prognosis analysis of patients with spinal metastases treated by percutaneous vertebroplasty combined with postoperative radiotherapy and radiotherapy alone].

Objective: To evaluate the efficacy of percutaneous vertebroplasty(PVP) combined with postoperative radiotherapy and radiotherapy alone in the treatment of spinal metastatic tumors and to evaluate the prognostic factors for survival. Methods: From December 2011 to December 2015, according to the choice of treatment, patients in group A(60 cases) were treated with PVP combined with postoperative radiotherapy and those in group B(50 cases) underwent radiotherapy alone, age, sex, primary tumor type , and other basic characteristics were analyzed in both groups in department of orthopedics and radiotherapy department, 307 Hospital of the People's Liberation Army. The pain visual analogue scale(visual analogue scale, VAS), tumors of the spine instability score(the spinal instability neoplastic score and sins), physical status score(Karnofsky performance score and KPS) were used to evaluate pain, spinal stability improvement and physical condition. Kaplan-Meier was used to analyze the survival rates of two groups of patients and the influence of primary tumor types on the survival of patients; Cox proportional hazard model was used to calculate the correlations between survival and visceral metastases, system medical treatment, vertebral number before treatment and physical condition. Results: There was no significant difference in baseline data between the two groups(P>0.05). The VAS in the group A was significantly lower than the scores in the group B at 1 month, 3 months, 6 months, and 12 months after surgery. The SINS score dropped from(7.8±1.2) to(6.3±0.9)(1 month), (6.1±0.8)(3 months) in patients with PVP combined with postoperative radiotherapy(P<0.05), the SINS score of radiotherapy patients simply dropped from(7.6±0.9) to(7.4±0.7)(1 month), (7.3±0.6)(3 months), and there was no statistically significant difference(P=0.12). The survival rates of 6 months, 1 years, and 3 years were similar between two groups(P>0.05). The influence of different types of primary tumors on the survival time of the patients was statistically significant(P<0.05). Multiple analysis showed that the internal organs metastasis, systemic medical treatment, the number of vertebral bodies and the physical condition were the important prognostic factors of the survival in patients with spinal metastases. Conclusion: PVP combined with postoperative radiotherapy for spinal metastases is better than radiotherapy alone in the treatment of relieving pain, maintaining the stability of vertebral body and improving the quality of life of patients. Survival prognosis was similar in two groups. The types of primary tumors, visceral metastasis, systemic medical treatment, the number of vertebral bodies and the physical condition are important prognostic factors in the survival of patients with spinal metastases.

Alkaline extracellular reduction: isolation and characterization of an alkaliphilic and halotolerant bacterium, Bacillus pseudofirmus MC02.

AIMS: To isolate an alkaliphilic bacterium and to investigate its ability of extracellular reduction. METHODS AND RESULTS: An alkaliphilic and halotolerant humus-reducing anaerobe, Bacillus pseudofirmus MC02, was successfully isolated from a pH 10·0 microbial fuel cell. To examine its ability of extracellular reduction, AQDS (anthraquinone-2, 6-disulfonae), humic acids (HA) and Fe(III) oxides were chosen as representative electron acceptors. All the experiments were conducted in a pH 9·5 carbonate buffer. The results are as follows: (i) Sucrose, lactate, glucose and glycerol were the favourable electron donors for AQDS reduction by the strain MC02; (ii) The strain had the ability of reducing HA in the presence of sucrose; (iii) It could effectively reduce Fe(III) oxides coupled with sucrose fermentation when AQDS was added as electron shuttle and its Fe(III) reducing capacity ranked as: lepidocrocite (γ-FeOOH) > goethite (α-FeOOH) > haematite(α-Fe(2) O(3) ); (iv) The strain could decolourize azo dye Orange I. CONCLUSIONS: Bacillus pseudofirmus MC02 was capable of extracellular reduction in AQDS, HA and Fe(III) oxides, and it can be used for decolourizing azo dye (Orange I) in alkaline conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of an alkaliphlic strain of B. pseudofirmus capable of extracellular reduction in AQDS, HA, Fe(III) oxides and decolourization of Orange I. This study could provide valuable information on alkaline biotransformation in the printing and dyeing wastewater and saline-alkali soil.

Enhanced reductive dechlorination of DDT in an anaerobic system of dissimilatory iron-reducing bacteria and iron oxide.

The transformation of DDT was studied in an anaerobic system of dissimilatory iron-reducing bacteria (Shewanella decolorationis S12) and iron oxide (alpha-FeOOH). The results showed that S. decolorationis could reduce DDT into DDD, and DDT transformation rate was accelerated by the presence of alpha-FeOOH. DDD was observed as the primary transformation product, which was demonstrated to be transformed in the abiotic system of Fe(2+)+alpha-FeOOH and the system of DIRB+alpha-FeOOH. The intermediates of DDMS and DBP were detected after 9 months, likely suggesting that reductive dechlorination was the main dechlorination pathway of DDT in the iron-reducing system. The enhanced reductive dechlorination of DDT was mainly due to biogenic Fe(II) sorbed on the surface of alpha-FeOOH, which can serve as a mediator for the transformation of DDT. This study demonstrated the important role of DIRB and iron oxide on DDT and DDD transformation under anaerobic iron-reducing environments.

Fe(III) oxide reduction and carbon tetrachloride dechlorination by a newly isolated Klebsiella pneumoniae strain L17.

AIMS: To isolate an iron-reducing bacterium and examine its ability of Fe(III) oxide reduction and dechlorination. METHODS AND RESULTS: A fermentative facultative anaerobe, strain L17 isolated from subterranean sediment, can reduce Fe(III) oxides and carbon tetrachloride (CT). It was identified as Klebsiella pneumoniae by 16S rRNA sequence analysis. Strain L17 can metabolize fermentable substrates such as citrate, glycerol, glucose and sucrose coupled with the reduction of hydrous ferric oxide, goethite, lepidocrocite and hematite. Fe(III) reduction was influenced by crystal structure of Fe(III) oxide, type of fermentable substrate, metabolic status of the strain, and significantly enhanced by addition of anthraquinone-2,6-disulfonate (AQDS). Strain L17 could dechlorinate CT to chloroform, and the rate was accelerated in the presence of Fe(III) oxide and AQDS. Biotic dechlorination by strain L17 and abiotic dechlorination by sorbed Fe(II) were proposed as the two main mechanisms. AQDS might accelerate the dechlorination by transferring electrons from strain L17 to Fe(III) oxide and CT. CONCLUSIONS: K. pneumoniae L17 can reduce Fe(III) oxides and CT. The two reductions can occur simultaneously, and be significantly promoted by AQDS. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of a strain of K. pneumoniae capable of reducing Fe(III) oxides and CT. As a strain of environmental origin, strain L17 may have the potential for bioremediation of chlorinated compound-contaminated sites.

Horse embryonic stem cell lines from the proliferation of inner cell mass cells.

Inner cell mass (ICM) cells were isolated immunosurgically from day 7-8 horse blastocysts and, after proliferation in vitro for 15-28 passages, three lines of cells were confirmed to be embryonic stem (ES) cells by their continued expression of alkaline phosphatase activity and their ability to bind antisera specific for the recognized stem cell markers, SSEA-1, TRA-1-60, TRA-1-81, and the key embryonic gene Oct-4. When maintained under feeder cell-free conditions in vitro, the three lines of cells differentiated into cells of ectodermal, endodermal, and mesodermal lineages. However, they did not form teratomata when injected into the testes of severe combined immunodeficiency (SCID)/beige immunoincompetent mice, thereby indicating a significant difference in phenotype between ES cells of the horse and those of the mouse and human.