Objective To study the inhibitory effect and its mechanism of sodium butyrate on human laryngeal carcinoma in nude mice. Methods Human laryngeal carcinoma cell line Hep-2 was seeded in the subcutaneous layer of 12 nude mice to built laryngeal carcinoma xenograft model. Then they were randomly and equally divided into 2 groups. Sodium butyrate was given in experimental group while phosphatic-buffered saline (PBS) was used in control group for 4 weeks. Tumor size and body weight of the mice were measured at regular time-intervals. The tumor,heart,liver,lungs,spleen and kidneys were removed at the end of treatment. Tumor sections were examined by electronic microscopy. TUNEL method and immunohistochemical S-P method were used for detecting the expression of Ki-67 nuclear antigen and survivin protein. The heart,liver,lung,spleen and kidney sections were examined after HE staining for assessment of toxicity. Results In experimental group,the volume of tumors was reduced,the area of necrosis in tumors was widened,the apoptotic rate was increased obviously and the expression level of Ki-67 nuclear antigen and survivin protein was decreased as compared with control group. During treatment,all the nude mice grew well and there were no toxic reactions. At the end of treatment,there were no abnormal changes in heart,liver,lung,spleen and kidney sections examined under light microscope. Conclusion Sodium butyrate can significantly inhibit the growth of human laryngeal carcinoma xenograft in nude mice. Its mechanism may be related to the apoptosis in tumor cells by inhibiting the expression of survivin protein and Ki-67 nuclear antigen. There is no toxicity to heart,liver,lungs,spleen and kidneys at a treatment dose of sodium butyrate.

Objective To study the effect of the location and volume of tumors on the normal lung dose-volume parameters for lung cancer. Methods An sphere with a diameter of 2 cm made of tissue-equiv-alent material used for simulating tumor was inserted into the superior lobe, middle lobe, inferior lobe of the right lung, and superior lobe, inferior lobe of the left lung of the Rando phantom, respectively. 5-field sIM-RT plans were designed. The prescribed dose was 60 Gy/2 Gy/30 f, and 99% of the planning target volume received this dose. Dose-volume parameters of normal lung tissue including V_5, V_(10), V_(20), V_(30), V_50 and mean lung dose were analyzed and compared. Results For the dose-volume parameters, the diameter and the po-sition of the tumor have a significant effect (P < 0.05). With the diameter expanding from 2 cm to 3 cm,the parameters associated with tumor in various lobes increased by a range between 3.83%-125.38%,while the parameters linked with tumor in different lobes increased by a range between 10.46%-51.46% with the diameter expanding from 3 cm to 4 cm. Conclusions Location and diameter of sphere-like tumor have obvious effect on dose-volume parameters. Knowing about the degree of influence will help oncologists and physicists to evaluate treatment planning better and reduce radiation pneumonitis.

Objective:To compare bone marrow-sparing intensity-modulated radiotherapy(BMS-IMRT)with conventional (four-field box[3DCRT]and anteroposterior-posteroanterior[CRT])techniques in the treatment of cervical cancer.Methods:For a cohort of 10 patients,BMS-IMRT,3DCRT and CRT planning were designed.The prescribed dose was 45Gy/1.8Gy/25f,95%of the planning target volume received this dose.Doses were computed with a commercially available TPS.Plans were compared according to dose-volume histogram (DVH)analysis in terms of PTV homogeneity and conformity indices(HI and CI)as well as OARs dose and volume parameters.Results:BMS-IMRT had an advantages over 3DCRT and CRT in terms of CI,but inferior to the latter two for HI.BMS-IMRT was superior to 3DCRT in reducing the dose to PBM,small bowel,bladder and rectum.Compared with CRT,BMS-IMRT reduced the volume irradiated to the doses from 30Gy to 40Gy,but increased the volume irradiated to the low doses from 5Gy to 20Gy.In addition,BMS-IMRT reduced the volume of small bowel,bladder,rectum at nearly all dose levels.Conclusion:BMS-IMRT reduced irradiation of PBM compared with 3DCRT technique.Compared with CRT technique,BMS-IMRT reduced the volume of PBM irradiated to high doses.Therefore,for patients with cervical cancer after hysterectomy,BMS-IMRT might reduce acute hematologic toxicity(HT)compared with conventional techniques.

Objective To compare bone marrow-sparing intensity-modulated radiotherapy (BMS-IMRT) with conventional intensity-modulated radiotherapy (IMRT) without considering pelvic bone marrow (PBM) as a planning constraint in the treatment of cervical cancer after hysterectomy. Methods BMS-IM-RT and IMRT planning were separately designed in a cohort of 10 patients with cervical cancer after hysterec-tomy. The prescribed dose was 95% planning target volume receiving 45 Gy/25 f. A commercially available TPS with convolution/superposition (CS) algorithm was used for dose calculation. Plans were compared ac-cording to dose-volume histogram (DVH) analysis in terms of PTV homogeneity (HI), conformity index (CI) as well as dose and volume parameters of organ at risks (OARs). Results BMS-IMRT was better than IMRT in terms of CI, but inferior to the latter for HI. When compared with IMRT, V_5, V_(10), V_(20), V_(30) and V_(40) of PBM in BMS-IMRT were reduced by 1.81% ,8.61% ,31.81% ,29.50% and 28.29%, respec-tively. No statistically significant differences were found between BMS-IMRT and IMRT for dose distritutions of the small bowel, bladder or rectum. Conclusions For patients with cervical cancer after hysterectomy, BMS-IMRT can reduce the PBM volume irradiated by low dose, which may reduce acute hematologic toxici-ties.

Objective: To compare inverse three-dimensional conformal radiotherapy (Inv 3D-CRT) and intensity modulated ra-diotherapy (IMRT) for non-small cell lung cancer. Methods: For a cohort of 10 patients, Inv 3D-CRT and three groups of IMRT plannings were designed for per patient. The prescribed dose was 60 Gy/2 Gy/30f, 95% of the planning target volume received this dose. Dose was computed with a commercially available TPS using convolution/superposition (CS) algorithm. Plans were compared according to the PTV_(95)V_(20) ratio (PTV_(95)V_(20)) and D_(max)-D_(min). Results: Compared with Inv 3D-CRT, the PTV_(95)V_(20) ratio of three groups of IMRT increased by 1.08 (P = 0.014), 0.72 (P = 0.089) and 0.42 (P = 0.318), respectively. Conclusions: For NSCLC, IMRT can reduce the dose to the lungs compared with inverse 3D-CRT by improving the conformity of the plan and is worth spreading in clinical work.

Objective:To investigate the effects of long noncoding RNA (LncRNA) lung cancer associated transcript 1 (LUCAT1) targeting microRNA (miR)-502-5p on the proliferation, migration and invasion of human retinoblastoma (RB) cells.Methods:RB tissue samples were collected from 27 RB patients who underwent eyeball enucleation in Henan Eye Hospital from May 2019 to January 2021.Another 27 normal retinal tissue specimens were collected from 12 patients with eyeball rupture, 7 with eyeball atrophy, and 8 with eyeball penetrating injury combined with pigment film incarceration who underwent the eyeball enucleation in Henan Eye Hospital during the same period.The expressions of LncRNA LUCAT1 and miR-502-5p in RB tissues, cell lines (Y-79, WERI-Rb-1, HXo-RB44) and human retinal epithelial cells (ARPE-19) were detected by real-time quantitative PCR.Y-79 RB cell was divided into control group, small interfering RNA (si)-LncRNA LUCAT1 group, si-control (con) group, pcDNA group, pcDNA-LncRNA LUCAT1 group, miR-con group, miR-502-5p group, si-LncRNA LUCAT1+ anti-miR-con group and si-LncRNA LUCAT1+ anti-miR-502-5p group, and cells in different groups were transfected with corresponding reagents.The expressions of MMP2 and MMP9 proteins were detected by Western blot.Cell proliferation activity was assayed by cell counting kit 8.Cell proliferation capability was detected by colony formation assay.Cell migration and invasion ability were determined by Transwell assay.The targeting regulation of LncRNA LUCAT1 against miR-502-5p was confirmed by dual luciferase reporter assay and real-time quantitative PCR.The study protocol was approved by the Ethics Committee of Henan Eye Hospital (No.HNEECKY-2021[32]). Written informed consent was obtained from guardians of subjects.Results:LncRNA LUCAT1 expression level in RB tissue was 2.73±0.34, which was significantly higher than 1.00±0.15 in normal retinal tissue ( t=24.190, P<0.001). The miR-502-5p expression level in RB tissues was 0.42±0.06, which was significantly lower than 1.00±0.13 in normal retinal tissue ( t=21.049, P<0.001). LncRNA LUCAT1 expression level was significantly higher and the miR-502-5p expression level was significantly lower in human RB cell lines Y-79, WERI-Rb-1 and HXO-RB44 than those in ARPE-19 cells, with statistically significant differences (all at P<0.05). The LncRNA LUCAT1 expression, the relative expressions of MMP2 and MMP9 proteins, the absorbance ( A) value, and the number of proliferated, migrating and invading Y-79 cells in si-LncRNA LUCAT1 group were significantly reduced in comparison with control group, and the differences were statistically significant (all at P<0.05). The miR-502-5p expression level was higher, and the relative expression levels of MMP2 and MMP9, A value, as well as the number of proliferated, migrating and invading Y-79 cells were lower in miR-502-5p group than in miR-con group, showing statistically significant differences ( t=20.274, 14.884, 14.181, 12.692, 17.749, 20.889, 21.913; all at P<0.001). The miR-502-5p expression level was lower and the relative expression levels of MMP2 and MMP9, A value as well as the number of proliferated, migrating and invading Y-79 cells were higher in si-LncRNA LUCAT1+ anti-miR-502-5p group than in si-LncRNA LUCAT1+ anti-miR-con group, showing statistically significant differences ( t=14.097, 15.839, 15.757, 11.860, 16.235, 16.565, 16.487; all at P<0.001). When co-transfected with LncRNA LUCAT1-wild type, the relative luciferase activity of miR-502-5p group was lower than that of miR-con group, and the difference was statistically significant ( t=16.379, P<0.001). The LncRNA LUCAT1 expression level was higher and the miR-502-5p expression level was lower in pcDNA-LncRNA LUCAT1 group than in pcDNA group, and the differences were statistically significant (both at P<0.05). The LncRNA LUCAT1 expression level was lower and the miR-502-5p expression level was higher in si-LncRNA LUCAT1 group than in si-con group, and the differences were statistically significant (both at P<0.05). Conclusions:Inhibition of LncRNA LUCAT1 can attenuate the proliferation, migration and invasion ability of human RB cells by the targeting up-regulation of miR-502-5p.

Objective:To screen for the mutation types of the FOXL2 in 4 families with blepharophimosis-ptosis-epicanthus inversus syndrome (BPES), and explore their genotype-phenotype correlations.Methods:To retrospectively analyze the result of FOXL2 gene detection by fluorescence quantification PCR, Sanger sequencing and multiplex ligation-dependent probe amplification(MLPA) in probands and families of BPES from January 2018 to January 2021, obtain mutation sites of pathogenic genes.Results:4 BPES families (8 cases) including 4 males and 4 females, with age ranged form 4 to 52 years (mean 24). The proband of family 1 has fragment deletion of Chr3∶138, 944, 224-138, 947, 137 region of FOXL2 gene. Proband of family 2 has heterozygosity deletion upstream of the 5’end of FOXL2 gene. There are missense mutations of c. 241 T>C(p.Y81 H)in proband and affected mother of family 3. There are c. 672_701dup(p.A224_A234dup) in-frame duplication mutations in proband and affected mother of family 4.Conclusions:Identification of causative mutations in the BPES patients has provided a basis for genetic counseling and reproductive guidance. The fragment deletion of Chr3∶138, 944, 224-138, 947, 137 region of FOXL2 gene and heterozygosity deletion upstream of the 5’end of FOXL2 gene are all new mutations that have not been reported. The novel mutations have enriched the mutation spectrum of the FOXL2 gene.

Objective:To screen for the mutation types of the FOXL2 in 4 families with blepharophimosis-ptosis-epicanthus inversus syndrome (BPES), and explore their genotype-phenotype correlations.Methods:To retrospectively analyze the result of FOXL2 gene detection by fluorescence quantification PCR, Sanger sequencing and multiplex ligation-dependent probe amplification(MLPA) in probands and families of BPES from January 2018 to January 2021, obtain mutation sites of pathogenic genes.Results:4 BPES families (8 cases) including 4 males and 4 females, with age ranged form 4 to 52 years (mean 24). The proband of family 1 has fragment deletion of Chr3∶138, 944, 224-138, 947, 137 region of FOXL2 gene. Proband of family 2 has heterozygosity deletion upstream of the 5’end of FOXL2 gene. There are missense mutations of c. 241 T>C(p.Y81 H)in proband and affected mother of family 3. There are c. 672_701dup(p.A224_A234dup) in-frame duplication mutations in proband and affected mother of family 4.Conclusions:Identification of causative mutations in the BPES patients has provided a basis for genetic counseling and reproductive guidance. The fragment deletion of Chr3∶138, 944, 224-138, 947, 137 region of FOXL2 gene and heterozygosity deletion upstream of the 5’end of FOXL2 gene are all new mutations that have not been reported. The novel mutations have enriched the mutation spectrum of the FOXL2 gene.

Blood-brain barrier (BBB) damage after ischemia significantly influences stroke outcome. Compound LFHP-1c was previously discovered with neuroprotective role in stroke model, but its mechanism of action on protection of BBB disruption after stroke remains unknown. Here, we show that LFHP-1c, as a direct PGAM5 inhibitor, prevented BBB disruption after transient middle cerebral artery occlusion (tMCAO) in rats. Mechanistically, LFHP-1c binding with endothelial PGAM5 not only inhibited the PGAM5 phosphatase activity, but also reduced the interaction of PGAM5 with NRF2, which facilitated nuclear translocation of NRF2 to prevent BBB disruption from ischemia. Furthermore, LFHP-1c administration by targeting PGAM5 shows a trend toward reduced infarct volume, brain edema and neurological deficits in nonhuman primate