Objective To explore the effects of homocysteine (Hcy) on neural stem cell (NSC) proliferation and the mRNA expression level of Notch1 and Hes1 related Notch signaling pathway from neonatal rats in vitro. Methods NSCs from neonatal rats were cultured by serum-free culture method in vitro. Cells were divided into four groups: control group (Hcy-C), low dose Hcy (Hcy-L) group, middle dose Hcy (Hcy-M) group and high dose Hcy (Hcy-H) group. NSCs were iden- tified by immunofluorescent staining using the antibodies against Nestin, β-tubulin Ⅲ and GFAP. The proliferation ability of NSCs was detected by MTT. The mRNA expressions of Notch1 and Hes1 were detected by Real-time PCR. Results In the serum free suspension medium, neurospheres that consisted of a great number of nestin-positive cells were found. β-tu- bulin Ⅲ positive neurons and GFAP positive astrocytes were detected by immunofluorescence staining on the 6 th day of cell induction. MTT assay showed that the cell viability was significantly lower in three Hcy treatment groups than those of con- trol group (P ＜ 0.05). And the effect of concentration-dependent was observed. The results of RT-PCR showed that mRNA expression of Hes1 was significantly lower in three Hcy treatment groups than that in control group (P ＜ 0.05). The mRNA ex- pression of Notch1 was significantly lower in Hcy-H group than that of other three groups (P ＜ 0.05). The mRNA expression of Notch1 was significantly lower in Hcy-M group than that of Hcy-L group and control group (P ＜ 0.05). Conclusion Hcy could inhibit the proliferation of NSCs by down-regulating mRNA expression levels of Notch1 and Hes1 genes related to Notch signal pathway.

Prostate cancer is one of the most common malignant tumors in men and related studies have achieved great breakthrough in recent years.But because of the lack of effective in vivo animal models, the process to translate basic research into clinical application has been severely hampered.Patient derived prostate tumor xenograft ( PDPTX) model is an ideal animal model in which freshly isolated tumor tissues from patients were inoculated into immunodeficient mice.This model can duplicate the heterogeneity of primary tumor in a better way and keep the tumor complexity at molecular, genetic and pathological levels.Particularly, the PDPTX model, in which the isolated tumor tissue is inoculated under the renal capsule, is even better, because it solves the clrawbacks of traditional subcutaneous inoculation model.In traditional mod-els, the success rate is low, it’s not easy for lower grade tumor to form xenograft, and it’s not easy to reconstruct metasta-sis, etc.PDPTX provides a more ideal in vivo model for prostate cancer studies.It has irreplaceable advantages, especially in target therapy, new drug screening and individualized tumor treatment.

The core-shell nanopaticles of Au@polyvinyl-pyrrolidone ( PVP) with uniform size and controllabe shell-thickness were prepared by hydrothermal method. The core-shell nanoparticles could be assembled to be the monolayer array on Si substrate relying on the dispersion of core-shell nanoparticles arising from PVP shell. The malachite green ( MG ) absorbed by H-bond could be detected on the array under the electromagnetic enhancement of inner-core Au nanoparticles. Under the conditions of the optimum shell-thickness of Au@PVP and the appropriate absorbed time of MG, the detection of MG could be realized in the linear range from 1 × 10-10 mol/L to 1 × 10-5 mol/L with the correlation coefficient ( R2 ) of 0. 98. The detection limit was 10-12 mol/L. This method was applied to the determination of MG in tilapia fish fillets of Xiagang market. No MG was found in this real sample. The spiked recoveries of the sample ranged from 70. 8% to 126. 0%. This method is simple and accurate, and can be used for detection of MG in the fish.

ObjectiveTo assess the differences of short- and long-term postintervention status on ocular and systemic symptoms for patients with ocular myasthenia gravis (OMG) after pyridostigmine bromide, corticosteroid, thymectomy, or thymectomy-corticosteroid combination therapy ( combination ).MethodsThis retrospective plus prospective study included 180 OMG patients, whose age of onset ≥ 15 years, treated non-randomly with above therapies separately: thymectomy group (60 cases ), corticosteroid group (39 cases), combination group ( 31 cases ), symptomatic group ( 50 cases ). Postintervention status complying with Myasthenia Gravis Foundation of America (MGFA)complete stable remission ,pharmacologic remission, or minimal manifestations was considered as desirable response, which was used as statistical indicator. Results ①Corticosteroid group showed higher desirable response rates on ptosis, ophthalmoplegia and general weakness at 3-6 months after treatment than other groups, and 42. 1％( 16/38 ) of them at 3 months achieved the desired state of ptosis, higher than the symptomatic group (7/48,14. 6％, ×2 = 8. 200, P = 0. 004 ). ② Ascending ideal rates had been presented in both combination and thymectomy groups since 1 year after treatments, while a little bit higher rate was presented in the former. At the end of observation, 21.7％ ( 13/60 )of patients in thymectomy group achieved complete stable remission.By paired longitudinal comparisons,thymectomy group showed higher ideal rates on ptosis (22/40,55.0％ ), ophthalmoplegia ( 16/27,59. 3％ ) and general weakness (20/40,50. 0％ ) at 2 years than that at 3 months( 11/59,18.6％ ;11/44,25.0％ ;9/60,15.0％ ;P =0. 002, 0. 031,0.000). ③For those patients by symptomatic treatment, the average age of onset was (51.9 ± 18.0) years, higher than that by other 3 therapies (F = 10. 563 ,P =0. 000). ④OMG patients with ophthalmoplegia more likely select corticosteroid or combined therapy. Ophthalmoplegia in combination group was higher than that in symptomatic and surgery groups( ×2 = 12. 939,14. 380, P =0. 000 in both). Ophthalmoplegia in corticosteroid group was higher than that in surgery group ( ×2 = 8. 017, P = 0. 005 ).Conclusions Corticosteroid appears to early overcome ptosis, ocular motor dysfunction and general weakness for patient with OMG in early-to-middle adulthood.Thymectomy andsurgery-corticosteroid combinationtherapies bothshowlong-term effectonthem.

Objective To explore the changes in expression of Klotho, an aging-suppression protein, and neutrophil gelatinase associated lipocalin ( NGAL) and their relationship with rat mesangial cells ( RMCs) cultured with high glucose in vitro, and to explore the role played by Toll-like receptor-4 (TLR4) / nuclear factor-kB(NF-kB) p65 pathways in this process. Methods Three NGAL-siRNA sequences were designed and synthesized. The effective sequence in subsequent experiments was chosen. RMCs were preincubated with pyrrolidinedithiocarbamate (PDTC)or exogenously added Klotho prior to high glucose treatment. Realtime PCR was used to analyze Klotho, TLR4, NGAL mRNA expressions. Western blot was used to observe Klotho, TLR4,NF-kB p65, NGAL,fibronectin (FN), and connective tissue growth factor ( CTGF) protein expression. ELISA assay was used to detect monocyte chemoattractant protein-1 ( MCP-1) and CXCL5 secretions. Results High glucose suppressed Klotho expression significantly(P<0. 05) and activated TLR4 / NF-kB p65 pathway. Meanwhile,the levels of NGAL,FN,CTGF, MCP-1, and CXCL5 were highly expressed ( P < 0. 01). NGAL gene silencing obviously down-regulated the increased expressions of FN, CTGF, MCP-1, and CXCL5 ( P < 0. 01). After PDTC treatment the overexpression of NGAL protein was markedly lowered(P<0. 01). In addition, Klotho treatment significantly inhibited the activity of TLR4 /NF-kB p65 pathways and down-regulated the expressions of NGAL, FN, CTGF, MCP-1 and CXCL5 stimulated by high glucose(P<0. 01). Conclusion Klotho inhibits the activity of TLR4 / NF-kB p65 pathways and thus inhibits NGAL expression in RMCs cultured with high glucose in vitro. And then it suppresses the expressions of FN, CTGF, MCP-1, and CXCL5. This provides a new basis to illustrate the protection mechanism of the anti-aging protein Klotho in diabetic nephropathy, and may provide new ideas and therapeutic targets for prevention and treatment.

Objective To explore thc clinical manifestation of secondary generalized myasthenia gravis(GMG) and analyze the factors affecting the progression from ocular myasthenia gravis(OMG) to GMG.Methods This research constitutes a single-center,retrospectively-collected prospective cohort study.We comprehensively reviewed our self-managed myasthenia gravis (MG) database drawn from personal clinical experience from January 2000 to Junc 2013.Patients underwent series of examination including repetitive nerve stimulation (RNS) tests,measurement of serum acetylcholine receptors antibody and serum muscle-specific tyrosine kinase antibodies,thymus computer tomography scan etc.Patients were treated with pyridostigmine bromide,corticosteroid therapy and (or) thymectomy based on a nonrandomization pattern and they were documented for their respective symptoms of OMG and GMG and date of GMG conversion.Logistic regression analysis was adopted to determine the influencing factors correlated with the development of GMG during the follow-up.Results Totally 770 patients initially diagnosed with OMG were included,among whom 573 (74％) patients remained with OMG (R-OMG group) and 197(26％) patients developed into GMG (GMG group) during the follow-up.(1) In comparison with their R-OMG counterparts,patients with secondary GMG were older at onset; Displayed more frequent RNS abnormality of facial nerve,accessory nerve and ulnar nerve ; Showed higher incidence of thymoma and were less treated by early corticosteroids.(2) For GMG group,81％ (160/197) of them displayed bulbar MG; 67％ (132/197) of GMG conversion occurred within 2 years,and 84％ (166/197) within 5 years.In comparison with the patients with onset of≤ 14-year-old,both of patients with15-49-year-old and≥ 50-year-old displayed higher conversion rate and shorter conversion duration (median:10 years versus 1 year and 6.5 months).(3) RNS abnormality of accessory nerve(OR =6.650,95％ CI 3.547-12.471 ; P ＜ 0.05) and thymoma(OR =7.924,95％ CI 2.554-24.585 ; P ＜ 0.05) were prognostic factors for the development of GMG,while early corticosteroid(OR =0.232,95％ CI 0.119-0.452 ; P ＜ 0.05) predicted the reduction of the risk of generalization.Conclusions Multiple factors including abnormal RNS of proximal limb muscles,thymoma,early corticosteroids therapy and possibly even onset age of over 15-year-old may involve the generalization in patients with OMG at onset.

[ ABSTRACT] The splicing factors were characterized for their crucial roles in pre-mRNA splicing of eukaryons. SRSF2 is a member of the SR protein family which is one of the most common splicing factors, and it is believed to be a key element in pre-mRNA splicing, mRNA transcription, regulation of the DNA stability and cell proliferation.SRSF2 gene mutation is detected frequently in myeloid malignancies ( like MDS and CMML) and may be associated with the phenotype and prognosis of these malignancies.The paper makes a review for the latest research progression on SRSF2 gene mutation and its relationship with myeloid malignancies.

With aligned MR registration, the MR-Linac provides superior soft tissue resolution for prostate cancer. No fiducial markers or electromagnetic transponders insertion is needed to guarantee high-precision radiotherapy. The highly-recommended Adapt-To-Shape (ATS) workflow can resolve all the problems encountered during prostate cancer radiotherapy, including prostate volume changes and adjacent organs motion, both inter-fractionally and intra-fractionally. With all the above advantages, MR-Linac performs outstandingly than conventional linac in prostate cancer RT delivery, and probably helps us to reduce the CTV-PTV margin safely in the near future. Nevertheless, it is difficult to implement the ATS workflow in clinical practice. In this article, the standard ATS workflow for prostate cancer was summarized based on our own experience.

Objective:To elucidate the change of whole genome expression profile for the effect of melatonin on radiation-induced intestinal injury in mice.Methods:C57BL/6J male mice were administrated with melatonin at 10 mg/kg body weight by intraperitoneal injection once a day for five consecutive days before abdominal irradiation with 14 Gy of γ-rays. Small intestines were harvested 3 d after radiation. GO annotation and KEGG pathway of the differential genes involved in small intestine were explored by DNA microarray analysis.Results:Compared with the control group, 584 differential genes were up-regulated and 538 differential genes were down-regulated for administration group pre-irradiation. The overlapping differential genes were selected from the irradiated mice and the administrated mice pre-irradiation. There were 324 up-regulated genes and 246 down-regulated genes unique to the administrated mice pre-irradiation. GO annotation analysis of the differential genes indicated that the top 15 significantly enriched biological processes for the administrated mice pre-irradiation mainly included autophagosome assembly (GO: 0000045), autophagosome organization (GO: 1905037) and regulation of acute inflammatory response (GO: 0002673). The genes ATG12, ATG16L2 and AMBRA1 were involved in autophagosome assembly and autophagosome organization. The genes C3, CPN1, CD55, CFP, CNR1, C1QA, C2 and CREB3L3 were involved in the regulation of acute inflammation response. KEGG pathway analysis of the differential genes involved indicated that the top 15 significantly enriched pathways for the administrated mice pre-irradiation mainly included O-glycan biosynthesis (hsa00512), glycosphingolipid biosynthesis (hsa00603), ECM-receptor interaction (hsa04512) and biosynthesis of unsaturated fatty acids (hsa01040). qRT-PCR verification showed that the expressions of ATG12 and ATG16L2 genes involved in autophagy for the administrated mice pre-irradiation increased significantly compared with the irradiated mice ( t=2.40, 4.35, P<0.05). Conclusions:The differential genes related with the biological process of autophagy, acute inflammatory response and the pathway of unsaturated fatty acid biosynthesis might be involved in the effect of melatonin on radiation-induced intestinal injury.

Objective:To investigate the workflow, efficacy and safety of MR-Linac in liver malignancies.Methods:Clinical data of 15 patients with hepatocellular carcinomas (HCC) or liver metastases treated with MR-Linac between November 2019 and July 2021 were retrospectively analyzed. The workflow of MR-Linac was investigated and image identification rate was analyzed. Patients were followed up for response and toxicity assessment.Results:Fifteen patients (6 HCC, 8 liver metastases from colorectal cancer, 1 liver metastasis from breast cancer) were enrolled. A total of 21 lesions were treated, consisting of 10 patients with single lesion, 4 patients with double lesions and 1 patient with triple lesions. The median tumor size was 2.4 cm (0.8-9.8 cm). The identification rate for gross tumor volume (GTV) in MR-Linac was 13/15. Although GTV of two patients were unclearly displayed in MR-Linac images, the presence of adjacent blood vessel and bile duct assisted the precise registration. All the patients were treated with stereotactic body radiation therapy (SBRT). For HCC, the median fraction dose for GTV or planning gross tumor volume (PGTV) was 6 Gy (5-10 Gy) and the median number of fractions was 9(5-10). The median total dose was 52 Gy (50-54 Gy) and the median equivalent dose in 2 Gy fraction (EQD 2Gy) at α/ β= 10 was 72 Gy (62.5-83.3 Gy). For liver metastases, the median fraction dose for GTV or PGTV was 5 Gy (5-10 Gy) and the median number of fractions was 10(5-10). The median total dose was 50 Gy (40-50 Gy) and the median EQD 2Gy at α/ β=5 was 71.4 Gy (71.4-107.1 Gy). At 1 month after SBRT, the in-field objective response rate (ORR) was 8/13 and the disease control rate was 13/13. At 3-6 months after SBRT, the in-filed ORR was increased to 6/6. During the median follow-up of 4.0 months (0.3-11.6), 4-month local progression-free survival, progression-free survival and overall survival were 15/15, 11/15 and 15/15, respectively. Toxicities were mild and no grade 3 or higher toxicities were observed. Conclusions:MR-Linac provides a platform with high identification rates of liver lesions. Besides, the presence of adjacent blood vessel and bile duct also assists the precise registration. It is especially suitable for liver malignancies with promising local control and well tolerance.