[EBF1 Promotes the Sensitivity of Cervical Cancer Cells to Cisplatin via Activating FBN1 Transcription].

Cisplatin (DDP) is widely used in the chemotherapy of cervical cancer (CC), the fourth most common female malignancy worldwide. However, some patients progress to chemotherapy resistance, which leads to chemotherapy failure, tumor recurrence, and poor prognosis. Therefore, strategies to identify the regulatory mechanisms underlying CC development and increase tumor sensitivity to DDP will help improve patient survival. This research was designed to ascertain the mechanism of EBF1-dependent regulation of FBN1 which promotes chemosensitivity of CC cells. The expression of EBF1 and FBN1 was measured in CC tissues resistant or sensitive to chemotherapy and in DDP-sensitive or -resistant cells (SiHa and SiHa-DDP cells). SiHa-DDP cells were transduced with lentiviruses encoding EBF1 or FBN1 to evaluate the influence of these two proteins on cell viability, expression of MDR1 and MRP1, and cell aggressiveness. Moreover, the interaction between EBF1 and FBN1 was predicted and demonstrated. Finally, to further verify the EBF1/FB1-dependent mechanism of DDP sensitivity regulation in CC cells a xenograft mouse model of CC was established using SiHa-DDP cells transduced with lentiviruses carrying EBF1 gene and shRNA directed to FBN1 EBF1 and FBN1 showed decreased expression in CC tissues and cells, particularly in those resistant to chemotherapy. Transduction of SiHa-DDP cells with lentiviruses encoding EBF1 or FBN1 lead to decreased viability, IC50, proliferation capacity, colony formation ability, aggressiveness, and increased cell apoptosis. We have shown that EBF1 activates FBN1 transcription by binding to FBN1 promoter region. Additionally, it was revealed that FBN1 silencing reversed the promoting effect of EBF1 overexpression on chemosensitivity of CC cells in vivo. EBF1 facilitated chemosensitivity in CC cells by activating FBN1 transcription.

Small intestinal morphology and sugar transporters expression when consuming diets of different energy levels: comparison between Tibetan and small-tailed Han sheep.

Some non-structural carbohydrates, especially starch, escape ruminal fermentation, are converted into glucose, and are absorbed from the small intestine. This glucose provides an important source of energy, and its usage is more efficient than glucose from carbohydrates which are fermented as short chain fatty acids in the rumen and, subsequently, undergo hepatic gluconeogenesis. Tibetan sheep graze on the harsh Qinghai-Tibetan Plateau (QTP) all year round and their carbohydrate and energy intakes fluctuate greatly with seasonal forage availability. Consequently, a high capacity to absorb glucose from the small intestine would be particularly beneficial for Tibetan sheep to allow them to cope with the inconsistent dietary intakes. This study examined how the small intestinal morphology and sugar transporters' expression of Tibetan and Small-tailed Han (Han) sheep respond to fluctuating energy intakes under the harsh conditions of the QTP. Han sheep graze on the QTP only in summer and are generally raised in feedlots. Twenty-four Tibetan sheep and 24 Han sheep, all wethers, were assigned randomly to four groups (n = 6 per breed/group), with each group offered a diet differing in digestible energy content: 8.21, 9.33, 10.45 and 11.57 MJ/kg DM. After 49 d, all sheep were slaughtered, tissues of the small intestine were collected, and measurements were made of the morphology and glucose transporters and the related regulation gene expressions. At intakes of low energy levels, Tibetan sheep had a greater villus surface area in the duodenum, jejunum and ileum and higher mRNA expression of sodium-dependent glucose transporter 1 in the duodenum and ileum (P < 0.05) than Han sheep. In the glucose transporter 2 (GLUT2) mediated glucose absorption pathway, Tibetan sheep had higher GLUT2 and taste receptor family 1 member 2 and 3 mRNA expressions than Han sheep in the duodenum, jejunum and ileum (P < 0.05). We concluded that the differences between breeds indicated a greater glucose absorption capacity in the small intestine of Tibetan than Han sheep, which would confer an advantage to Tibetan over Han sheep to an inconsistent energy intake on the harsh QTP. These findings suggested that ruminants raised under harsh environmental conditions with highly fluctuating dietary intakes, as is often the case in grazing ruminants worldwide, are able to absorb glucose from the small intestine to a greater extent than ruminants raised under more moderate conditions.

Tazarotene gel promotes healing of deep tissue injury in mice.

We investigated the efficacy and molecular mechanisms of tazarotene gel for healing deep tissue injury (DTI). We used male C57BL/6J mice to establish a DTI model. Animals were divided randomly into control, tazarotene gel and purilon gel groups. We injected 100 ul tazarotene gel, purilon gel or saline every 48 h for 20 days. Hematoxylin and eosin staining was used to observe pathological changes on days 14 and 21. The mRNA and protein expression of VEGF-α, TGF-ß1 and HIF-1α were detected by qRT-PCR and western blot, respectively. Wound sites exhibited accelerated healing by 20 days in the tazarotene gel group. Fewer inflammatory cells and more granulation tissue were found in both experimental groups compared to controls. The mRNA and protein expression of VEGF-α and TGF-ß1 in the experimental groups were increased compared to the control group by day 14. Expression of HIF-1α in the experimental groups was significantly less than in the controls. Tazarotene gel promoted wound healing independent of the HIF-1α/VEGF signalling pathway during tissue repair of DTI. Tazarotene and purilon gels exhibited similar macroscopic healing of wounds and expression of genes and proteins.

[Efficacy and safety of endoscopic papillectomy of major duodenal papilla neoplasms].

Objective: To discuss the efficacy and safety of endoscopic papillectomy of major duodenal papilla neoplasms. Methods: The clinical-pathological data of 21 patients who were admitted to the Department of Endoscopy, Cancer Hospital, Chinese Academy of Medical Sciences and underwent endoscopic papillectomy of major duodenal papilla neoplasms from January 2014 to January 2020 were retrospectively studied, their postoperative outcomes and complication were also analyzed. Results: Tweenty-one patients were successfully performed endoscopic papillectomy of major duodenal papilla neoplasms. The resected lesions varied between 0.5-2.8 cm. Completed lesion was resected in 19 cases and lesion blocks in 2 cases. The incidence of postoperative complication was 52.4% (11/21), including 8 cases of postoperative bleeding (38.1%). Five patients stopped bleeding after endoscopic hemostasis and 3 patients stopped after interventional embolization. Two patients experienced perforation (9.5%) and recovered after conservative treatment including anti-inflammatory treatment and abdominal drainage. Five patients had pancreatitis (23.8%) and recovered after treatment with pre-somatostatin and anti-inflammatory rectal suppository. Preoperative pathological results of 21 patients suggested that 11 were high-grade intraepithelial neoplasia and 8 were low-grade intraepithelial neoplasia, and 2 were chronic inflammation. Postoperative pathological results suggested that 4 were adenocarcinoma, and the rest 17 were adenoma. The coincidence rate of preoperative biopsy results and postoperative pathology was 38.1%(8/21), and underestimate of the pathological stage occurred in 11 patients (52.4%) during the preoperative biopsy, overestimate occurred in two patients (9.5%). Four cases had a positive incisal margin. All patients had good prognoses and no death event occurred during the follow-up period. Conclusions: Early-stage major duodenal papilla neoplasms should be treated with aggressive resection. Endoscopic papillectomy of duodenal papilla neoplasms is safe, effective, and can be recommended as the preferred procedure for major duodenal papilla neoplasms.

Energy substrate metabolism in skeletal muscle and liver when consuming diets of different energy levels: comparison between Tibetan and Small-tailed Han sheep.

The energy intake of Tibetan sheep on the harsh Qinghai-Tibetan Plateau (QTP) varies greatly with seasonal forage fluctuations and is often below maintenance requirements, especially during the long, cold winter. The liver plays a crucial role in gluconeogenesis and skeletal muscle is the primary tissue of energy expenditure in mammals. Both play important roles in energy substrate metabolism and regulating energy metabolism homeostasis of the body. This study aimed to gain insight into how skeletal muscle and liver of Tibetan sheep regulate energy substrate metabolism to cope with low energy intake under the harsh environment of the QTP. Tibetan sheep (n = 24; 48.5 ± 1.89 kg BW) were compared with Small-tailed Han sheep (n = 24; 49.2 ± 2.21 kg BW), which were allocated randomly into one of four groups that differed in dietary digestible energy densities: 8.21, 9.33, 10.45 and 11.57 MJ /kg DM. The sheep were slaughtered after a 49-d feeding period, skeletal muscle and liver tissues were collected and measurements were made of the activities of the key enzymes of energy substrate metabolism and the expressions of genes related to energy homeostasis regulation. Compared with Small-tailed Han sheep, Tibetan sheep exhibited higher capacities of propionate to glucose conversion and fatty acid oxidation and ketogenesis in the liver, higher glucose utilization efficiency in both skeletal muscle and liver, but lower activities of fatty acid oxidation and protein mobilization in skeletal muscle, especially when in negative energy balance. However, the Small-tailed Han sheep exhibited higher capacities to convert amino acids and lactate to glucose and higher levels of glycolysis and lipogenesis in the liver than Tibetan sheep. These differences in gluconeogenesis and energy substrate metabolism conferred the Tibetan sheep an advantage over Small-tailed Han sheep to cope with low energy intake and regulate whole-body energy homeostasis under the harsh environment of the QTP.

[Application of ultrasound-guided hip joint drug injection in the postoperative rehabilitation of arthroscopie repair of acetabular labral tears].

OBJECTIVE: To explore the clinical application of ultrasound-guided hip joint drug injection in the postoperative rehabilitation of arthroscopie repair of acetabular labral tears. METHODS: This research retrospectively analyzed a total of 38 hips from 36 patients (2 of them were bilateral) whose imaging examination showed acetabular labral well healed but the rehabilitating training was limited due to hip pain after arthroscopie repair of acetabular labral tears in our hospital between June 2015 and May 2017. All the patients underwent ultrasound-guided hip joint drug injection treatment. Through comparing the pain and the function of hip before and after drug injection, the clinical application values of ultrasound-guided hip joint drug injection in the postoperative rehabilitation of arthroscopie repair of acetabular labral tears were explored. The degree of hip pain was assessed by visual analogue score (VAS), which were scored before and after the injection. The hip function was assessed by the hip range of activity. The SPSS 21.0 statistical software was used for the data analysis. The effective rate of hip injection was calculated, which was defined as: ("excellent" + "good")/total number of cases×100%. The degree of hip pain was assessed by VAS, which was divided into 0 to 10 points with 0 for no pain and 10 for unbearable severe pain. The function of hip was assessed by the hip range of activity. The therapeutic effect of "excellent" meant no pain or occasional slight pain in the hip, along with Patrick test was negative and hip joint was not limited; the therapeutic effect of "good" meant that the pain was significantly reduced, and the hip's activity was slightly restricted. "No effect" meant that the pain of hip was not relieved, and the Patrick test was positive. RESULTS: The VAS score of the patient before drug injection was 5.46±1.46, and the VAS score was 2.01±0.53 after drug injection 4 weeks later. The score of the latter was significantly lower than that of the former, and the difference was statistically significant (P<0.05). The hip joint activity after ultrasound-guided hip joint drug injection was significantly improved. The therapeutic effective rate was 84.2%. CONCLUSION: For patients with hip pain and limitations after arthroscopie repair of acetabular labral tears, ultrasound-guided drug injection can effectively reduce hip pain, improve hip activity, and promote hip functional reconstruction.

The Diagnostic Performance of Urine Flow Cytometer UF1000i for Urinary Tract Infections.

BACKGROUND: Urinary tract infections (UTIs) are one of the most common infectious diseases in clinic. Urine flow cytometry is receiving more and more attention due to its rapid forecast of UTIs. METHODS: The Urine Flow Cytometer UF1000i has a series of software programs to quantify bacteria (BACT) and white blood cells (WBC), and describe the scatter diagram of bacteria. The UTIs were predicted based on the cutoff values with the Receiver Operating Characteristic (ROC) curves of BACT and WBC counts. To evaluate the diagnostic performance of UF1000i for UTIs, the sensitivity and specificity of 889 urine samples were determined in comparison to the results of urine culture. Meanwhile the bacterial morphology indication of the UF1000i was evaluated in order to help doctors choose antibiotics. The angle of the scatter cloud with the x-axis was used to classify the infected bacteria as bacilli (< 30°) or cocci (≥ 30°). RESULTS: The best cutoff value of BACT counts for predicting UTIs was 119 per µL, and the sensitivity and specificity were 95.5% and 88.7%, respectively. While the best cutoff value of WBC counts was 81.5 per µL, and the sensitivity and specificity were 77.6% and 76.7%, respectively. In addition, the best cutoff values for females were 583 BACT per µL and 137.5 WBC per µL. They were much higher than for males (118 BACT per µL and 91 WBC per µL). The coincidence of the bacterial morphology information between the UF1000i software indication and the bacterial actual morphology identified by urine culture was 83% (bacilli) and 68% (cocci), respectively. CONCLUSIONS: Data demonstrated that the performance of BACT counts for UTIs is superior to WBC counts. In addition, the bacterial morphology could preliminarily be predicated by the scatter diagram. Since the urine flow cytometer UF1000i can provide the data of both BACT counts and the scatter diagram, the urine flow cytometry was regarded as a suitable method for screening UTIs. Moreover, it would be better to take gender into consideration when setting the best cutoff value for diagnosis of UTIs in clinic.

The effectiveness of transport in a toy car for reducing preoperative anxiety in preschool children: a randomised controlled prospective trial.

BACKGROUND: This study was designed to determine whether transport of a paediatric inpatient in a children's ride-on toy car has an effect on perioperative levels of anxiety compared with transport on a hospital gurney with or without oral midazolam premedication. METHODS: In this prospective study, 108 children aged 2-5 yr with congenital heart disease and undergoing first surgical correction were randomly allocated to one of three groups: Group C (transport in a children's ride-on car), Group G (transport on a gurney without premedication), or Group M (transport on a gurney and received premedication of oral midazolam 0.5 mg kg-1). The modified Yale Preoperative Anxiety Scale-Short Form and parent-recorded anxiety VAS were applied to evaluate anxiety in the following time points: pre-anaesthesia visit (the day before surgery), upon getting in the ride-on car or on the gurney in the ward, upon arriving in the preoperative holding area, at the moment of leaving from the holding area to the operating room (OR) (coincided with separation from parents), at the time after entering the OR, and at the time just before anaesthesia induction. RESULTS: Children in Group C exhibited significantly lower levels of anxiety from the time they got into the ride-on car until the time they entered the OR, compared with the other two groups (P<0.001). The subjects in Group C had similarly low anxiety levels to those in the Group M at the time before induction (P=0.914). CONCLUSIONS: Transport in a ride-on toy car can relieve preoperative anxiety in preschool children undergoing surgery to a comparable degree as midazolam. CLINICAL TRIAL REGISTRATION: ChiCTR-IOR-17012791.

The overexpression of Fra1 disorders the inflammatory cytokine secretion by mTEC of myasthenia gravis thymus.

The thymus of a myasthenia gravis (MG) patient is often accompanied by and effected with follicular hyperplasia. Inflammatory cytokines in thymus induce the formation of germinal centres (GC). MG thymic inflammatory cytokines are predominantly secreted by stromal cells. Our previous studies revealed that the expression level of the Fra1 protein, which is a Fos member of the activator protein 1 transcription factors (AP-1), was higher in the MG thymus compared with that of the normal thymus. Based on that, we demonstrated that Fra1 was mainly expressed in medulla thymic epithelial cells (mTECs) and that the rate of Fra1 positive mTECs in the MG thymus was higher than normal. In vitro, we found that the expression of CCL-5, CCL-19 and CCL-21 could be regulated by Fra1 in mTEC and that IL-1ß, IL-6, IL-8 and ICAM1 were downregulated in the Fra1 overexpression group and upregulated in the Fra1 knock-down group. Meanwhile, we detected that the expression levels of suppressor of cytokine signalling 3 (SOCS3) were significantly upregulated along with the overexpression of Fra1. Hence, we considered that the overexpression of Fra1 disrupted inflammatory cytokine secretion by mTEC in the MG thymus and that STAT3 and SOCS3 were strongly involved in this process.

[Investigation of pregestational diabetes mellitus in 15 hospitals in Guangdong province].

Objective: To investigate the morbidity, diagnostic profile and perinatal outcome of pregestational diabetes mellitus (PGDM) in 15 hospitals in Guangdong province. Methods: A total of 41 338 women delivered in the 15 hospitals during the 6 months, 195 women with PGDM (PGDM group) and 195 women with normal glucose test result (control group) were recruited from these tertiary hospitals in Guangdong province from January 2016 to June 2016. The morbidity and diagnostic profile of PGDM were analyzed. The complications during pregnancy and perinatal outcomes were compared between the two groups. In the PGDM group, pregnancy outcomes were analyzed in women who used insulin treatment (n=91) and women who did not (n=104). Results: (1) The incidence of PGDM was 0.472%(195/41 338). Diabetes mellitus were diagnosed in 59 women (30.3%, 59/195) before pregnancy, and 136 women (69.7%,136/195) were diagnosed as PGDM after conceptions. Forty-six women (33.8%) were diagnosed by fasting glucose and glycohemoglobin (HbA1c) screening. (2) The maternal age, pre-pregnancy body mass index (BMI) , prenatal BMI, percentage of family history of diabetes, incidence of macrosomia, concentration of low density lipoprotein were significantly higher in PGDM group than those in control group (all P<0.05). Women in PGDM group had significantly higher HbA1c concentration ((6.3±1.3)% vs (5.2±0.4)%) , fasting glucose [(6.3±2.3) vs (4.8±1.1) mmol/L], oral glucose tolerance test (OGTT) -1 h glucose ((12.6±2.9) vs (7.1±1.3) mmol/L) and OGTT-2 h glucose [(12.0±3.0) vs (6.4±1.0) mmol/L] than those in control group (P<0.01). (3) The morbidity of preterm births was significantly higher (11.3% vs 1.0%, P<0.01), and the gestational age at delivery in PGDM group was significantly smaller [(37.6±2.3) vs (39.2±1.2) weeks, P<0.01]. Cesarean delivery rate in the PGDM group (70.8% vs 29.7%) was significantly higher than the control group (P<0.01). There was significantly difference between PGDM group and control in the neonatal male/female ratio (98/97 vs 111/84, P=0.033). The neonatal birth weight in PGDM group was significantly higher ((3 159±700) vs (3 451±423) g, P<0.01) . And the incidence of neonatal hypoglycemia in the PGDM group was higher than the control group (7.7% vs 2.6%, P=0.036). (4) In the PGDM group, women who were treated with insulin had a smaller gestational age at delivery [(36.9±2.9) vs (37.9±2.5) weeks, P<0.01], and the neonates had a higher neonatal ICU (NICU) admission rate (24.2% vs 9.6%, P<0.01). Conclusions: The morbidity of PGDM in the 15 hospitals in Guangdong province is 0.472%. The majority of PGDM was diagnosed during pregnancy; HbA1c and fasting glucose are reliable parameters for PGDM screening. Women with PGDM have obvious family history of diabetes and repeated pregnancy may accelerate the process of diabetes mellitus. Women with PGDM have higher risk for preterm delivery and neonatal hypoglycemia. Unsatisfied glucose control followed by insulin treatment may increase the need for NICU admission.

Elimination of chronic lymphocytic leukemia cells in stromal microenvironment by targeting CPT with an antiangina drug perhexiline.

Chronic lymphocytic leukemia (CLL) is the most common adult leukemia in the western countries and is currently incurable due, in part, to difficulty in eliminating the leukemia cells protected by stromal microenvironment. Based on previous observations that CLL cells exhibit mitochondrial dysfunction and altered lipid metabolism and that carnitine palmitoyltransferases (CPT) have a major role in transporting fatty acid into mitochondria to support cancer cell metabolism, we tested several clinically relevant inhibitors of lipid metabolism for their ability to eliminate primary CLL cells. We discovered that perhexiline, an antiangina agent that inhibits CPT, was highly effective in killing CLL cells in stromal microenvironment at clinically achievable concentrations. These effective concentrations caused low toxicity to normal lymphocytes and normal stromal cells. Mechanistic study revealed that CLL cells expressed high levels of CPT1 and CPT2. Suppression of fatty acid transport into mitochondria by inhibiting CPT using perhexiline resulted in a depletion of cardiolipin, a key component of mitochondrial membranes, and compromised mitochondrial integrity, leading to rapid depolarization and massive CLL cell death. The therapeutic activity of perhexiline was further demonstrated in vivo using a CLL transgenic mouse model. Perhexiline significantly prolonged the overall animal survival by only four drug injections. Our study suggests that targeting CPT using an antiangina drug is able to effectively eliminate leukemia cells in vivo, and is a novel therapeutic strategy for potential clinical treatment of CLL.

Runx1 contributes to neurofibromatosis type 1 neurofibroma formation.

Neurofibromatosis type 1 (NF1) patients are predisposed to neurofibromas but the driver(s) that contribute to neurofibroma formation are not fully understood. By cross comparison of microarray gene lists on human neurofibroma-initiating cells and developed neurofibroma Schwann cells (SCs) we identified RUNX1 overexpression in human neurofibroma initiation cells, suggesting RUNX1 might relate to neurofibroma formation. Immunostaining confirmed RUNX1 protein overexpression in human plexiform neurofibromas. Runx1 overexpression was confirmed in mouse Schwann cell progenitors (SCPs) and mouse neurofibromas at the messenger RNA and protein levels. Genetic inhibition of Runx1 expression by small hairpin RNA or pharmacological inhibition of Runx1 function by a Runx1/Cbfß interaction inhibitor, Ro5-3335, decreased mouse neurofibroma sphere number in vitro. Targeted genetic deletion of Runx1 in SCs and SCPs delayed mouse neurofibroma formation in vivo. Mechanistically, loss of Nf1 increased embryonic day 12.5 Runx1(+)/Blbp(+) progenitors that enable tumor formation. These results suggest that Runx1 has an important role in Nf1 neurofibroma initiation, and inhibition of RUNX1 function might provide a novel potential therapeutic treatment strategy for neurofibroma patients.

Upregulated expression of NKG2D and its ligands give potential therapeutic targets for patients with thymoma.

The activating receptor NKG2D (natural killer group 2, member D) of natural killer (NK) cells promotes tumor immune surveillance by targeting ligands selectively induced on cancer cells, and thus having an important role in antitumor immune response. Because these ligands are not widely expressed on healthy adult tissue, NKG2D ligands may present as useful target for immunotherapeutic approaches in cancer. In this study, to elucidate the role of NKG2D-NKG2D ligand interaction in thymoma tissues and to evaluate the potential role of NKG2D ligands as therapeutic target for thymoma, we examined the expression of NKG2D and its specific ligands: MICA (major histocompatibility complex class I chain-related protein A), MICB (major histocompatibility complex class I chain-related protein B) and ULBP (UL16-binding protein) in 36 thymomas (6 subtype A, 6 subtype AB, 8 subtype B1, 5 subtype B2, 6 subtype B3 and 5 subtype C), 15 thymic atrophy and 8 thymic hyperplasia by immunohistochemistry and reverse transcription-real-time-PCR methods. We demonstrated that both mRNA and protein levels of NKG2D, MICA, MICB and ULBP were upregulated in six types of thymomas compared with those in atrophic thymus or proliferating thymus. Furthermore, the NKG2D ligands were found to be frequently coexpressed on thymoma cells. Furthermore, the expression of MICA, MICB and ULBP in subtype C was higher compared with those in subtype A, AB, B1, B2 and B3. Thus, we concluded that high expressions of NKG2D, MICA, MICB and ULBP1 were shown in patients with thymoma, and this may enhance the recognition function of NK cells to eliminate tumor cells. MICA, MICB and ULBP presented an attractive target for thymoma therapy. The abnormal expression of NKG2D, MICA, MICB and ULBP1 can provide us with evidence of the occurrence of thymoma and could also be used as a target in the treatment of thymoma.

Runx1 is required for hematopoietic defects and leukemogenesis in Cbfb-MYH11 knock-in mice.

CBFß-SMMHC (core-binding factor ß-smooth muscle myosin heavy chain), the fusion protein generated by the chromosome 16 inversion fusion gene, CBFB-MYH11, is known to initiate leukemogenesis. However, the mechanism through which CBFß-SMMHC contributes to leukemia development is not well understood. Previously, it was proposed that CBFß-SMMHC acts by dominantly repressing the transcription factor RUNX1 (Runt-related protein 1), but we recently showed that CBFß-SMMHC has activities that are independent of RUNX1 repression. In addition, we showed that a modified CBFß-SMMHC with decreased RUNX1-binding activity accelerates leukemogenesis. These results raise questions about the importance of RUNX1 in leukemogenesis by CBFß-SMMHC. To test this, we generated mice expressing Cbfb-MYH11 in a Runx1-deficient background, resulting from either homozygous Runx1-null alleles (Runx1(-/-)) or a single dominant-negative Runx1 allele (Runx1(+/lz)). We found that loss of Runx1 activity rescued the differentiation defects induced by Cbfb-MYH11 during primitive hematopoiesis. During definitive hematopoiesis, RUNX1 loss also significantly reduced the proliferation and differentiation defects induced by Cbfb-MYH11. Importantly, Cbfb-MYH11-induced leukemia had much longer latency in Runx1(+/lz) mice than in Runx1-sufficient mice. These data indicate that Runx1 activity is critical for Cbfb-MYH11-induced hematopoietic defects and leukemogenesis.

Metabolic regulation of cancer cell side population by glucose through activation of the Akt pathway.

Side population (SP) cells within tumors are a small fraction of cancer cells with stem-like properties that can be identified by flow cytometry analysis based on their high ability to export certain compounds such as Hoechst 33342 and chemotherapeutic agents. The existence of stem-like SP cells in tumors is considered as a key factor contributing to drug resistance, and presents a major challenge in cancer treatment. Although it has been recognized for some time that tumor tissue niches may significantly affect cancer stem cells (CSCs), the role of key nutrients such as glucose in the microenvironment in affecting stem-like cancer cells and their metabolism largely remains elusive. Here we report that SP cells isolated from human cancer cells exhibit higher glycolytic activity compared to non-SP cells. Glucose in the culture environment exerts a profound effect on SP cells as evidenced by its ability to induce a significant increase in the percentage of SP cells in the overall cancer cell population, and glucose starvation causes a rapid depletion of SP cells. Mechanistically, glucose upregulates the SP fraction through ATP-mediated suppression of AMPK and activation of the Akt pathway, leading to elevated expression of the ATP-dependent efflux pump ABCG2. Importantly, inhibition of glycolysis by 3-BrOP significantly reduces SP cells in vitro and impairs their ability to form tumors in vivo. Our data suggest that glucose is an essential regulator of SP cells mediated by the Akt pathway, and targeting glycolysis may eliminate the drug-resistant SP cells with potentially significant benefits in cancer treatment.

First Report of a Tospovirus in Mulberry.

Mulberry (Morus alba L.) is an economically important crop grown widely throughout Asia. Various virus-like symptoms including mosaics, vein banding, and chlorotic ringspots have been observed and reported on mulberry trees in China and Japan for decades. However, the etiology of mulberry viral diseases is generally understudied, although two mulberry-infecting viruses, Mulberry latent virus (genus Carlavirus) (2) and Mulberry ringspot virus (genus Nepovirus) (3), have been partially characterized. In a recent (2010 to 2011) field survey in Guangxi Province, China, supported by the local government, the incidence of virus-like diseases of mulberry ranged between 40 and 80%. To identify the viruses infecting mulberry, deep sequencing of small RNAs (4) was conducted using an Illumina Genome Analyzer. Small RNAs were isolated from five samples of mulberry leaves showing various virus-like symptoms and sequenced. Among the contigs assembled, a 445-bp contig (GenBank Accession No. JX268597) was found to share 76.6% nucleotide identity and 83.0% amino acid identity to Groundnut bud necrosis virus (genus Tospovirus, family Bunyaviridae; Accession Nos. U42555 and AAC55521). To obtain a longer cDNA fragment of this virus, a reverse transcription (RT)-PCR was done with primers MV-N-F (5'-AAGCCATCAATGTGCCTCCGGA-3') and MV-N-R (5'-AACACCATGTCTACCGTCCGTC-3') that align to the S-RNA sequence encompassing the nucleocapsid (N) gene and a portion of the intergenic region (IGR) of the Tospovirus. PCR products of about 1,000 bp were successfully amplified from the total RNA of the three mulberry samples (sl-1, xcsy-1, and xcsy-4) showing vein banding symptoms, but not from asymptomatic mulberry (jk-1). These PCR products were cloned and sequenced. The lengths of the amplicons were 1,027 bp (isolate sl-1, JX173786), 987 bp (isolate xcsy-1, JX173787), and 979 bp (isolate xcsy-4, JX173788) and the partial IGRs of the sl-1, xcsy-1, and xcsy-4 isolates were 187 bp, 147 bp, and 139 bp, respectively. The coding regions for the N protein were 831 bp and the deduced proteins of 277 amino acid residues were 100% identical for all three isolates. Since the N protein of this virus shared up to only 74.4% identity to other tospoviruses (74.4% to Capsicum chlorosis virus, ABB83818; and 71.5% to Watermelon bud necrosis virus, ABY79095), it may represent a new member of the Tospovirus genus, temporarily named Mulberry vein banding virus (MuVBV), according to the species demarcation criteria for the Bunyaviridae (1). To the best of our knowledge, this is the first report of a Tospovirus infecting M. alba. In an RT-PCR screening of 48 randomly selected mulberry samples suspected to be virus-infected, 32 were MuVBV-positive. Giving the high incidence and the high yield loss associated with Tospovirus and the presence of thrips, suspected vectors for the virus, MuVBV may represent a substantial threat to the silkworm industry in China. References: (1) M. Q. K. Andrew et al. Virus Taxonomy: 9th Report of the ICTV. Elsevier Academic Press, San Diego, 2012. (2) T. Tsuchizaki. Annu. Phytopath. Soc. Japan 42:304, 1976. (3) T. Tsuchizaki et al. Annu. Phytopath. Soc. Japan 37:266, 1971. (4) Q. Wu et al. PNAS. 107:1606, 2010.

Autophagy activation is associated with neuroprotection against apoptosis via a mitochondrial pathway in a rat model of subarachnoid hemorrhage.

Autophagy, the bulk intracellular degradation of cytoplasmic constituents, can be a pro-survival or a pro-death mechanism depending on the context. A recent study showed that autophagy was activated in the phase of early brain injury following subarachnoid hemorrhage (SAH). However, whether autophagy activation after SAH is protective or harmful is still elusive. This study was undertaken to determine the potential role of autophagy pathway activation in early brain injury following SAH. The rats were pretreated with intracerebral ventricular infusion of either the autophagy inducer rapamycin (RAP) or inhibitor 3-methyladenine (3-MA) before SAH onset. The results from electron microscopic examinations showed that RAP administration caused the formation of autophagosomal vacuoles, and 3-MA induced neuronal apoptosis. RAP treatment significantly increased the expression of autophagic proteins Atg5 and Beclin 1, the ratio of microtubule-associated protein 1 light chain 3 (LC3)-II to LC3-I and reduced caspase-3 activity, the number of terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL)-positive cells, brain edema and neurological deficits after SAH. Conversely, 3-MA treatment exacerbated early brain injury. RAP treatment significantly increased the expression of the autophagic proteins Atg5 and Beclin 1, the ratio of LC3-II to LC3-I and reduced caspase-3 activity, the number of TUNEL-positive cells, brain edema and neurological deficits after SAH. Conversely, 3-MA treatment reversed these changes and exacerbated early brain injury. To further clarify the mechanism of autophagy protection, we investigated the expression levels of key apoptosis-related molecules. The results showed that RAP administration decreased Bax translocation to the mitochondria and downstream cytochrome c release from the mitochondria to the cytosol. Taken together, our study indicates that activation of autophagic pathways reduces early brain injury after SAH. This neuroprotective effect is likely exerted by anti-apoptotic mechanisms.