Analysis of clinical characteristics of thyroid phenotype in Pendred syndrome based on multiple databases.

OBJECTIVE: This study aimed to provide statistical data support for the development of thyroid phenotype-related follow-up and reference for follow-up duration and project selection by analyzing the clinical characteristics of thyroid phenotype in Pendred syndrome (PDS) based on multiple databases. MATERIALS AND METHODS: PDS-related pathogenic or possibly/pathogenic mutations were searched by Deafness Variation Database (DVD), ClinVar, and PubMed databases, the mutation sites were counted and the characteristics and thyroid phenotypes were analyzed. RESULTS: The median age of hearing phenotype onset in PDS cases reported in multiple databases was 1.0 (1.0, 2.0) years, the median age of thyroid phenotype onset was 14.5 (5.8, 21.0) years, and the median age that thyroid phenotype was more delayed than hearing phenotype was 10.0 (4.0, 17.0) years. There were significant differences in the distribution of onset time between the two phenotypes (Z=-4.560, p<0.01). In these patients, the positive rates of goiter, thyroid nodules, abnormal thyroid function, and perchlorate discharge test (PDT) were 78%, 78%, 69%, and 78%, respectively. Moreover, the number of thyroid phenotype-positive items in the genotype group with frameshift mutation was not significantly higher than that in the group without frameshift mutation (Z=-1.452, p=0.147). CONCLUSIONS: The early missed diagnosis of PDS may be due to the late onset of thyroid phenotype and the non-100% positive rate of examination items. Therefore, multi-item follow-up of the thyroid gland into adulthood will benefit patients. At present, the relationship between genotype and phenotype is still unclear, and prognosis cannot be determined according to genotype.

[Preliminary study on the expression of MIF in HCC tissues and its relationship with ERK1/2 signaling pathway].

Objective: To investigate the expression of Macrophage migration-inhibitory factors (MIF) in hepatocellular carcinoma (HCC) tissues and its interaction with ERK1/2 signaling pathway, so as to establish a theoretical basis for further studying the molecular mechanism of MIF promoting HCC. Methods: From February 2020 to August 2021, 52 cases of hepatocellular carcinoma (HCC) tissues based on hepatitis B cirrhosis (HBV-LC) and 52 cases of adjacent tissues in Tianjin Medical University Cancer Hospital and 940th Hospital of Joint Logistic Support Force of PLA were collected as the experimental group, including 39 males and 13 females, aged 35-65 years. And 20 cases of normal liver tissue were selected as the control group. Immunohistochemistry was used to detect the expression of MIF, ERK1/2 and p-ERK1/2 proteins in liver tissues of the two groups, and in situ hybridization was used to detect the expression of ERK1/2 nucleic acid in liver tissues of the two groups.HepG2 HCC cells and L-02 normal hepatocytes were co-cultured with different concentrations of rMIF, the expression and phosphorylation levels of ERK1/2 and JNK1 proteins in the two kinds of liver cells were detected by Western-blot, and the expression levels of ERK1/2 nucleic acids in the two kinds of liver cells were detected by RT-PCR. One-way ANOVA was used for measurement data and χ2 test was used for counting data. Results: The expressions of MIF, ERK1/2, p-ERK1/2 and ERK1/2 mRNA were significantly increased in HCC and para-cancer tissues (the expression of MIF in HCC group was 78.8%, and that in adjacent group was 75.0%; ERK1/2 80.8% in HCC group and ERK1/2 71.8% in paracancerous group. The expression of p-ERK1/2 75.0 % in HCC group and 46.2% in paracancerous group were respectively detected. ERK1/2 mRNA was expressed in HCC group 76.9%, ERK1/2 mRNA expression in paracancerous group 78.8%), and the differences were statistically significant compared with normal liver tissues (P<0.05), but there was no significant difference between HCC and para-cancer tissues (P>0.05). The expressions of ERK1/2, p-ERK1/2 and ERK1/2 mRNA in HepG2 HCC cells were significantly increased with the increase of rMIF concentration, and the increase was most obvious when rMIF concentration was 200 ng/ml, and the difference was statistically significant compared with L-02 normal hepatocytes (P<0.05). Conclusion: MIF, ERK1/2 and p-ERK1/2 are highly expressed in HCC tissues and HepG2 HCC cells, suggesting that MIF promotes the occurrence and development of hepatocellular carcinoma through ERK1/2 signaling pathway.

A 2-year locomotive exploration and scientific investigation of the lunar farside by the Yutu-2 rover.

The lunar nearside has been investigated by many uncrewed and crewed missions, but the farside of the Moon remains poorly known. Lunar farside exploration is challenging because maneuvering rovers with efficient locomotion in harsh extraterrestrial environment is necessary to explore geological characteristics of scientific interest. Chang'E-4 mission successfully targeted the Moon's farside and deployed a teleoperated rover (Yutu-2) to explore inside the Von Kármán crater, conveying rich information regarding regolith, craters, and rocks. Here, we report mobile exploration on the lunar farside with Yutu-2 over the initial 2 years. During its journey, Yutu-2 has experienced varying degrees of mild slip and skid, indicating that the terrain is relatively flat at large scales but scattered with local gentle slopes. Cloddy soil sticking on its wheels implies a greater cohesion of the lunar soil than encountered at other lunar landing sites. Further identification results indicate that the regolith resembles dry sand and sandy loam on Earth in bearing properties, demonstrating greater bearing strength than that identified during the Apollo missions. In sharp contrast to the sparsity of rocks along the traverse route, small fresh craters with unilateral moldable ejecta are abundant, and some of them contain high-reflectance materials at the bottom, suggestive of secondary impact events. These findings hint at notable differences in the surface geology between the lunar farside and nearside. Experience gained with Yutu-2 improves the understanding of the farside of the Moon, which, in return, may lead to locomotion with improved efficiency and larger range.

[A comparative study of transperitoneal transmesenteric approach versus paracolic sulci approach laparoscopic adrenal tumorectomy for treatment of primary hyperaldosteronism on left side].

Objective: To compare the therapeutic effect of transperitoneal transmesenteric approach versus paracolic sulci approach laparoscopic adrenal tumorectomy for treatment of left-sided primary hyperaldosteronism. Methods: From January 2017 to July 2019, the clinical data of 70 patients with left-sided primary hyperaldosteronism (PHA) who underwent surgery in the First Hospital of Lanzhou University and five other hospitals in Gansu Province were retrospectively analyzed. There are 43 male and 27 female patients. Among them,28 patients were performed transperitoneal transmesenteric approach laparoscopic adrenal tumorectomy and 42 patients were performed transperitoneal paracolic sulci approach laparoscopic adrenal tumorectomy. The general information and perioperative data of the two groups were compared. Results: All 70 cases of surgery were successfully completed. As compared with the paracolic sulci approach group, the operation time was significantly shorter in the transmesenteric approach group[(26.7±8.8)vs (38.9±7.1)min,P<0.001)], and the estimated blood loss was less in the transmesenteric approach group[45(30,50) vs 50(40,60)ml,P=0.042]. There was no statistically significant difference in the postoperative hospitalization days between the two groups[(4.4±1.0)vs(4.5±1.0)d, P=0.669)]. The electrolytes and aldosterone to renin ratio returned to a healthy level in the postoperative one month, and the blood pressure also returned to a healthy level in 53 (75.7%) patients. Conclusion: Transperitoneal transmesenteric approach laparoscopic adrenal tumorectomy is safe and feasible, with a short operation time and relatively less estimated blood loss.

[Analysis of bleeding risk in percutaneous renal biopsy in Tibet].

OBJECTIVE: To observe the postoperative bleeding after percutaneous renal biopsy (PRB) in Tibet, To analyze and summarize the risk factors associated with bleeding in high altitude patients to improve the safety of surgery. METHODS: A retrospective analysis of 150 cases of PRB in the Department of Nephrology, People's Hospital of Tibet Autonomous Region from May 2016 to May 2018 were carried out, and the correlations between the potential risk factors (gender, age, blood pressure, hemoglobin, platelet, serum creatinine) and postoperative bleeding events were analyzed. RESULTS: During the study period, the 150 patients receiving procedure of PRB were enrolled in our hospital, with an average age of (41.2±15.6) years, of whom 58.7% (88/150) were male, 41.3% (62/150) were female, and major bleeding complications occurred in 12 biopsies (8.0%, 12/150). Six cases for men and women, respectively. The mean age in the bleeding group seemed to be higher than that in the non-bleeding group [(48.3±20.0) years vs. (40.6±15.1) years, P=0.099]. There was no significant difference in the incidence of hypertension, hemoglobinemia, urea nitrogen and prothrombin time between the two groups. The level of serum creatinine in the hemorrhage group seemed to be higher than that in the non-bleeding group (P=0.090), and the time of the hemorrhagic group was longer than that in the non-bleeding group (P=0.069). The platelet count in the bleeding group was significantly lower than that in the non-bleeding group (P < 0.05). Multivariate Logistic regression analysis showed that the prolonged activation of partial prothrombin time and lower platelet count had a relatively high risk of bleeding, which was statistically significant (P=0.079, P=0.082). CONCLUSION: PRB is safe and reliable on the whole in plateau areas; Old age, low platelet count, decreased renal function and prolonged activated partial coagulation time are related to postoperative bleeding of PRB, and hyperhemoglobin is not a risk factor for bleeding. High hemoglobin is not a risk factor for postoperative bleeding of PRB at high altitude.

An epidemiological survey of echinococcosis in intermediate and definitive hosts in Qinghai Province, China.

In order to understand the epidemiological status of alveolar and cystic echinococcosis in intermediate and definitive hosts in Qinghai Province, China, during the period 2007-2011, we investigated the infection in humans and animals, including yaks, Tibetan sheep, Tibetan dogs, and wild foxes distributed in different counties around the province. Sera from local residents were examined using a rapid serodiagnostic kit to detect specific antibodies against Echinococcus. Seropositive samples were confirmed with B-scan ultrasonography and X-ray examinations. Yaks and Tibetan sheep were checked at slaughterhouses, and cysts and suspicious lesions were collected for analysis. A rapid diagnostic strip was used to detect Echinococcus adults in Tibetan dogs. Positive dogs were dewormed and the parasites collected. Wild foxes were trapped and necropsies performed with particular attention to the intestine. Forty-eight of 735 (6.4%) humans tested were positive and 475 of 854 (55.6%) Tibetan sheep and 85 of 352 (24.15%) yaks were infected with Echinococcus. Across different counties, 214 of 948 (22.57%) Tibetan dogs were positive, and five of 36 (13.9%) wild foxes were infected with Echinococcus. Molecular studies showed that all the infections detected in humans, domestic yaks, and Tibetan sheep were the G1 genotype (E. granulosus), whereas the parasites from Tibetan foxes and Tibetan dogs were E. shiquicus and E. multilocularis, respectively. In conclusion, Echinococcosis is hyperendemic in Qinghai Province in both its intermediate and definitive hosts and the G1 genotype of cystic Echinococcus is the dominant strain.

Limited fertility of the subcutaneous cysts of Echinococcus multilocularis.

Echinococcus multilocularis is a tiny devastating worm that causes alveolar echinococcosis in humans. This disease mainly occurs in the liver but rarely in other organs. We report the subcutaneous encystment of E. multilocularis metacestodes in experimentally infected mice. Subcutaneous cysts had remarkably fewer protoscoleces (2.05 ± 1.47, n = 20) and small irregular-shape vesicles (ISVs) in the lumen as compared to liver cysts (69.6 ± 55.65, n = 10). Moreover, abnormal development of a protoscolex was also observed in a subcutaneous cyst. The results suggest that subcutaneous encystment may have potential adverse effects on the reproductivity and development of protoscoleces, providing potential explanations for high tissue preference of metacestode encystment.

Design, fabrication and perivascular implantation of bioactive scaffolds engineered with human adventitial progenitor cells for stimulation of arteriogenesis in peripheral ischemia.

Cell therapy represents a promising option for revascularization of ischemic tissues. However, injection of dispersed cells is not optimal to ensure precise homing into the recipient's vasculature. Implantation of cell-engineered scaffolds around the occluded artery may obviate these limitations. Here, we employed the synthetic polymer polycaprolactone for fabrication of 3D woodpile- or channel-shaped scaffolds by a computer-assisted writing system (pressure assisted micro-syringe square), followed by deposition of gelatin (GL) nanofibers by electro-spinning. Scaffolds were then cross-linked with natural (genipin, GP) or synthetic (3-glycidyloxy-propyl-trimethoxy-silane, GPTMS) agents to improve mechanical properties and durability in vivo. The composite scaffolds were next fixed by crown inserts in each well of a multi-well plate and seeded with adventitial progenitor cells (APCs, 3 cell lines in duplicate), which were isolated/expanded from human saphenous vein surgical leftovers. Cell density, alignment, proliferation and viability were assessed 1 week later. Data from in vitro assays showed channel-shaped/GPTMS-crosslinked scaffolds confer APCs with best alignment and survival/growth characteristics. Based on these results, channel-shaped/GPTMS-crosslinked scaffolds with or without APCs were implanted around the femoral artery of mice with unilateral limb ischemia. Perivascular implantation of scaffolds accelerated limb blood flow recovery, as assessed by laser Doppler or fluorescent microspheres, and increased arterial collaterals around the femoral artery and in limb muscles compared with non-implanted controls. Blood flow recovery and perivascular arteriogenesis were additionally incremented by APC-engineered scaffolds. In conclusion, perivascular application of human APC-engineered scaffolds may represent a novel option for targeted delivery of therapeutic cells in patients with critical limb ischemia.

Distribution and diversity of bacterial communities and sulphate-reducing bacteria in a paddy soil irrigated with acid mine drainage.

AIMS: To investigate the effects of long-term acid mine drainage (AMD) irrigation on the change in bacterial community and sulphate-reducing bacteria (SRB) in a paddy soil. METHODS AND RESULTS: The bacterial community structures were investigated using 454 pyrosequencing, and 98 931 effective sequences were selected for the bacterial diversity analysis. The known dominant phyla in the paddy soil were Acidobacteria (33·5%), Proteobacteria (19·7%), Nitrospira (2·8%) and Actinobacteria (2·7%). Higher percentage of Acidobacteria than Proteobacteria was detected. The relative abundances of the dominant bacterial lineages were more significantly correlated with the soil pH, the organic matter and the sulphate than the heavy metals. The diversity of the SRB in the surface paddy soil showed that the uncultured SRB groups might play important roles in paddy soils. The other OTUs mainly belonged to six phylogenetic divisions: Desulfobacca, Desulfovibrio, Syntrophobacter, Desulforhopalus, Desulfarculus and Desulfobulbus. The distribution of the absolute abundance and the relative contribution of the SRB along the vertical soil profile were investigated by RT-PCR assays based on the dsrB gene. The abundance of the dsrB gene copy numbers was up to 1·92 × 10(9)  copies g(-1) dry soil, which is slightly higher than the other non-AMD-affected paddy soil. CONCLUSIONS: This study demonstrated that the abundance of SRB is increased by the AMD irrigation while changing the composition and diversity of the bacterial community in the paddy soil. SIGNIFICANCE AND IMPACT OF THE STUDY: This is, to our knowledge, the first attempt to characterize and quantify changes in the diversity and distribution of the microbial community and SRB in the long-term AMD-irrigated paddy soil, which will further increase our understanding of the impact of AMD on sulphur biogeochemical cycling in the paddy soil.

Fabrication of silicon molds with multi-level, non-planar, micro- and nano-scale features.

A method for single-step fabrication of arbitrary, complex, three-dimensional (3D) silicon structures from the nano- to millimeter-scale at multiple levels on non-planar, curved, or domed surfaces is reported. The fabrication is based on focused or masked ion beam irradiation of p-type silicon followed by electrochemical anodization. The process allows fabrication of a wide range of surface features at multiple heights and with arbitrary orientations by varying the irradiated feature width, ion type, energy fluence, and subsequent anodization conditions. The technology has achieved depth resolution of 10 nm as step heights and is capable of creating lateral features down to 7 nm at high aspect ratios of up to 40, with surface roughness down to 1 nm scaled up to full wafer areas. The single-step ability has seamlessly interfaced a network of complex, integrated micro- to nano-structures in 3D orientations with no alignment required. The final template has been converted to a master copy for nano-imprinting lithography of 3D fluidic structures and optical components.

Reprogramming hMSCs morphology with silicon/porous silicon geometric micro-patterns.

Geometric micro-patterned surfaces of silicon combined with porous silicon (Si/PSi) have been manufactured to study the behaviour of human Mesenchymal Stem Cells (hMSCs). These micro-patterns consist of regular silicon hexagons surrounded by spaced columns of silicon equilateral triangles separated by PSi. The results show that, at an early culture stage, the hMSCs resemble quiescent cells on the central hexagons with centered nuclei and actin/ß-catenin and a microtubules network denoting cell adhesion. After 2 days, hMSCs adapted their morphology and cytoskeleton proteins from cell-cell dominant interactions at the center of the hexagonal surface. This was followed by an intermediate zone with some external actin fibres/ß-catenin interactions and an outer zone where the dominant interactions are cell-silicon. Cells move into silicon columns to divide, migrate and communicate. Furthermore, results show that Runx2 and vitamin D receptors, both specific transcription factors for skeleton-derived cells, are expressed in cells grown on micropatterned silicon under all observed circumstances. On the other hand, non-phenotypic alterations are under cell growth and migration on Si/PSi substrates. The former consideration strongly supports the use of micro-patterned silicon surfaces to address pending questions about the mechanisms of human bone biogenesis/pathogenesis and the study of bone scaffolds.

Influence of the narrow {111} planes on axial and planar ion channeling.

We report channeling patterns where clearly resolved effects of the narrow {111} planes are observed in axial and planar alignments for 2 MeV protons passing through a 55 nm [001] silicon membrane. At certain axes, such as <213> and <314>, the offset in atomic rows forming the narrow {111} planes results in shielding from the large potential at the wide {111} planes, producing a region of shallow, asymmetric potential from which axial channeling patterns have no plane of symmetry. At small tilts from such axes, different behavior is observed from the wide and narrow {111} planes. At planar alignment, distinctive channeling effects due to the narrow planes are observed. As a consequence of the shallow potential well at the narrow planes, incident protons suffer dechanneled trajectories which are excluded from channeling within the wide planes, resulting in an anomalously large scattered beam at {111} alignment.

Relative parameter sensitivity in prenatal toxicity studies with substances classified as developmental toxicants.

Developmental toxicity testing according to the globally standardized OECD 414 protocol is an important basis for decisions on classification and labeling of developmental toxicants in the European Union (EU). This test requires relatively large animal numbers, given that parental and offspring generations are involved. In vitro assay designs and systems biology paradigms are being developed to reduce animal use and to improve prediction of human hazard. Such approaches could benefit from the long-term experience with animal protocols and more specifically from information on the relevance of effects observed in these tests for developmental toxicity. Therefore, we have analyzed relative parameter sensitivity in 22 publicly available developmental toxicity studies, representing about one third of all classified developmental toxicants under European legislation. Maternal and fetal weight effects and fetal survival were most often affected parameters at the developmental Lowest Observed Adverse Effect Level (dLOAEL), followed by skeletal malformations. Specific end points such as cleft palate were observed in fewer studies at dLOAEL, but if observed may have been crucial in classification and labeling decisions. These results are similar to earlier studies using different selections of chemicals, indicating that in general classified developmental toxicants have a similar pattern of effects at the dLOAEL as chemicals in general. These findings are discussed within the perspective of the development of innovative alternative approaches to developmental hazard assessment.

Construction of an artificial microalgal-bacterial consortium that efficiently degrades crude oil.

Four oil component-degrading bacteria and one oil-tolerant microalgae, Scenedesmus obliquus GH2, were used to construct an artificial microalgal-bacterial consortium for crude-oil degradation. The bacterial strains included Sphingomonas GY2B and Burkholderia cepacia GS3C, along with a mixed culture, named GP3, containing Pseudomonas GP3A and Pandoraea pnomenusa GP3B. GY2B could only degrade polycyclic aromatic hydrocarbons, GS3C was able to degrade aliphatic chain hydrocarbons, and GP3 could utilize both saturated and aromatic hydrocarbons. In combination with unialgal or axenic algae, the bacteria showed different effects on oil degradation. Unialgal GH2 was not suitable for the consortium construction, as it could not cooperate well with GS3C and GP3. The axenic GH2 exhibited no oil-degrading ability; however, it significantly promoted the degradation ability of the oil component-degrading bacteria, especially for degrading biorefractory polycyclic aromatic hydrocarbons. Axenic S. obliquus GH2, combined with the four bacteria mentioned above, formed an optimal algal-bacterial consortium. The artificial consortium demonstrated an elevated efficiency in degrading both aliphatic and aromatic hydrocarbons of crude oil.

Species differences for stereoselective metabolism of ethofumesate and its enantiomers in vitro.

1. The stereoselective metabolism of ethofumesate (ETO) and its enantiomers in rabbit and rat liver microsomes have been studied by chiral high-performance liquid chromatography (HPLC) method. Two metabolites were detected in both liver microsomes in the presence of beta-nicotinamide adenine dinucleotide phosphate (NADPH). 2. The T(1/2) of (+)-ETO and (-)-ETO in rabbit liver microsomes were 12.2 and 4.7 min of rac-ETO and 25.9 and 6.7 of ETO enantiomers. However, the T(1/2) of (+)-ETO and (-)-ETO in rat liver microsomes were 5.3 and 5.9 min of rac-ETO and 7.8 and 10.6 of ETO enantiomers. The stereoselective selectivity is similar to the in vivo study. 3. After incubation of ETO enantiomers, stereoselectivity was present in the formation of ETO-OH enantiomer in rabbit liver microsomes, but stereoselectivity was not evident in rat liver microsomes. 4. There was no chiral inversion from the (+)-ETO to (-)-ETO or inversion from (-)-ETO to (+)-ETO in both rabbit and rat liver microsomes.

Dose-dependent effects of soy phyto-oestrogen genistein on adipocytes: mechanisms of action.

The potential role of genistein in the prevention and treatment of obesity has attracted much attention among public and medical communities. Conversely, increasing evidence indicates that genistein as an endocrine-disrupting substance is likely to play a role in the aetiology of obesity. This review focuses on the role of soy phyto-oestrogen genistein in adipocytes and the underlying mechanisms of action. Genistein dose-dependently inhibits and stimulates adipogenesis in vitro. Increasing evidence shows that genistein dose-dependently influences obesity in both male and female animals. Dose-dependent effects of genistein on adipocytes vary with factors such as age and gender of animals. In addition, the role of developmental exposure of genistein in adult obesity has been discussed. Genistein, different from oestrogen, concurrently activates nuclear receptors, oestrogen receptors and peroxisome proliferator-activated receptors, and it inhibits various enzyme activities. The balance among these pleiotrophic effects of genistein determines its dose-dependent effects on adipocyte differentiation and function. Current data suggest that genistein could regulate adiposity. However, it remains uncertain whether genistein plays a beneficial role in the prevention and treatment of obesity. Additional evidence is required before firm conclusions showing that genistein decreases adiposity.

Cardiac contractile dysfunction in insulin-resistant rats fed a high-fat diet is associated with elevated CD36-mediated fatty acid uptake and esterification.

AIMS/HYPOTHESIS: Changes in cardiac substrate utilisation leading to altered energy metabolism may underlie the development of diabetic cardiomyopathy. We studied cardiomyocyte substrate uptake and utilisation and the role of the fatty acid translocase CD36 in relation to in vivo cardiac function in rats fed a high-fat diet (HFD). METHODS: Rats were exposed to an HFD or a low-fat diet (LFD). In vivo cardiac function was monitored by echocardiography. Substrate uptake and utilisation were determined in isolated cardiomyocytes. RESULTS: Feeding an HFD for 8 weeks induced left ventricular dilation in the systolic phase and decreased fractional shortening and the ejection fraction. Insulin-stimulated glucose uptake and proline-rich Akt substrate 40 phosphorylation were 41% (p < 0.001) and 45% (p < 0.05) lower, respectively, in cardiomyocytes from rats on the HFD. However, long-chain fatty acid (LCFA) uptake was 1.4-fold increased (p < 0.001) and LCFA esterification into triacylglycerols and phospholipids was increased 1.4- and 1.5-fold, respectively (both p < 0.05), in cardiomyocytes from HFD compared with LFD hearts. In the presence of the CD36 inhibitor sulfo-N-succinimidyloleate, LCFA uptake and esterification were similar in LFD and HFD cardiomyocytes. In HFD hearts CD36 was relocated to the sarcolemma, and basal phosphorylation of a mediator of CD36-trafficking, i.e. protein kinase B (PKB/Akt), was increased. CONCLUSIONS/INTERPRETATION: Feeding rats an HFD induced cardiac contractile dysfunction, which was accompanied by the relocation of CD36 to the sarcolemma, and elevated basal levels of phosphorylated PKB/Akt. The permanent presence of CD36 at the sarcolemma resulted in enhanced rates of LCFA uptake and myocardial triacylglycerol accumulation, and may contribute to the development of insulin resistance and diabetic cardiomyopathy.

At least two genetically distinct large Babesia species infective to sheep and goats in China.

A fatal disease of sheep and goats in the northern part of China has been reported to be due to Babesia ovis. However, some characteristics of the causative agent in recent reports are not in accordance with the original attributes ascribed to this parasite. Therefore, the 18S small subunit ribosomal RNA (18S rRNA) genes of a number of Babesia isolates in China were sequenced and compared with that of other Babesia and Theileria species in an attempt to clarify their taxonomic position. In the present study, seven Babesia isolates were collected from distinct areas of northern China, and the 18S rRNA genes were amplified and sequenced. The phylogenetic trees were inferred based on 18S rRNA gene sequences of the Chinese ovine Babesia isolates and some of ovine Babesia and Theileria species available in GenBank. In the phylogenetic tree, Babesia sp. isolates from Madang, Tianzhu, Lintan, Ningxian, Hebei and Liaoning all grouped with B. motasi with 88.2-99.9% identity, while Babesia sp. Xinjiang grouped in a separate clade between B. ovis and B. crassa with 79.7-81.2% identity. The results indicated that there are at least two distinct Babesia species groups-B. motasi and Babesia sp. Xinjiang, the latter was distinctly different from other ovine Babesia isolates from China with less than 86.6% identity.

Removal of serum factors by charcoal treatment promotes adipogenesis via a MAPK-dependent pathway.

In vitro differentiation of the progenitor cells or preadipocytes into adipocytes is usually achieved by adding an adipogenic mixture (isobutylmethylxanthine, dexamethasone, and insulin, IDI) to medium supplemented with fetal bovine serum (FBS). To study the effects of steroid hormones in vitro, endogenous hormones, growth factors and cytokines are removed by charcoal stripping of serum. However, the effects of charcoal-stripped serum (CS-FBS) per se on adipogenesis have been ignored. Here, we showed that alkaline phosphate activity and nodule formation of osteoprogenitor KS483 cells were lower in CS-FBS than in FBS. Concurrently, abundant amounts of adipocytes were only observed in KS483 cells cultured with CS-FBS, irrespective of the brands of serum used. Inhibition of the p42/44 MAPK pathway by its specific inhibitor PD98059 increased adipogenesis of KS483 cells with FBS, whereas activation of this signalling pathway by EGF blocked adipogenesis of these cells with CS-FBS. Furthermore, the p42/44 MAPK phosphorylation of KS483 cells cultured with CS-FBS was decreased compared with FBS. We concluded that charcoal-stripping of serum removed stimulators of the MAPK signalling pathway and in turn led to downregulation of osteogenesis and upregulation of adipogenesis. Interestingly, the adipogenic mixture IDI stimulated adipogenesis of KS483 cells cultured with CS-FBS, but not with FBS. Furthermore, differential effects of genistein on adipogenesis were observed in KS483 cells cultured with FBS or CS-FBS in combination with IDI. Our results showed that charcoal stripping of serum affected the commitment of KS483 cells and therefore differentially regulated adipogenesis influenced by IDI alone and in combination with genistein.

Exposure of KS483 cells to estrogen enhances osteogenesis and inhibits adipogenesis.

Osteoblasts and adipocytes arise from a common progenitor cell in bone marrow. Whether estrogen directly regulates the progenitor cells differentiating into osteoblasts or adipocytes remains unknown. Using a mouse clonal cell line KS483 cultured in charcoal-stripped fetal bovine serum (FBS), we showed that 17beta-estradiol (E2) stimulates the differentiation of progenitor cells into osteoblasts and concurrently inhibits adipocyte formation in an estrogen receptor (ER)-dependent way. E2 increased alkaline phosphate (ALP) activity and nodule formation and stimulated messenger RNA (mRNA) expression of core-binding factor alpha-1 (Cbfa1), parathyroid hormone/parathyroid hormone-related protein receptors (PTH/PTHrP-Rs), and osteocalcin. In contrast, E2 decreased adipocyte numbers and down-regulated mRNA expression of peroxisome proliferator-activated receptor-gamma (PPARgamma)2, adipocyte protein 2 (aP2), and lipoprotein lipase (LPL). Furthermore, the reciprocal control of osteoblast and adipocyte differentiation by E2 was observed also in the presence of the adipogenic mixture of isobutylmethylxanthine, dexamethasone, and insulin. Immunohistochemical staining showed that ERalpha and ERbeta were present in osteoblasts and adipocytes. A new mouse splice variant ERbeta2 was identified, which differed in two amino acid residues from the rat isoform. E2 down-regulated mRNA expression of ERalpha, ERbeta1, and ERbeta2. The effects of E2 are not restricted to the KS483 cell line because similar results were obtained in mouse bone marrow cell cultures. Our results indicate that estrogen, in addition to stimulation of osteogenesis, inhibits adipogenesis, which might explain the clinical observations that estrogen-deficiency leads to an increase in adipocytes.