AIM:To identify transcriptional differences between the ocular surface ectoderm(OSE)and surface ectoderm(SE)using RNA-seq, and elucidate the OSE transcriptome landscape and the regulatory networks involved in its development.METHODS:OSE and SE cells were differentiated from human embryonic stem(hES)cells. Differentially expressed genes(DEGs)between OSE and SE were analyzed using RNA-seq. Based on the DEGs, we performed gene ontology(GO)analysis, Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis, and protein-protein interaction(PPI)network analysis. Transcription factors(TFs)and hub genes were screened. Subsequently, TF-gene and TF-miRNA regulatory networks were constructed using the NetworkAnalyst platform.RESULTS:A total of 4 182 DEGs were detected between OSE and SE cells, with 2 771 up-regulated and 1 411 down-regulated genes in OSE cells. GO-BP analysis revealed that up-regulated genes in OSE were enriched in the regulation of ion transmembrane transport, axon development, and modulation of chemical synaptic transmission. Down-regulated genes were primarily involved in nuclear division, chromosome segregation, and regulation of cell cycle phase transition. KEGG analysis indicated that up-regulated genes in OSE cells were enriched in signaling pathways such as cocaine addiction, axon guidance, and amphetamine addiction, while down-regulated genes were enriched in proteoglycans in cancer, ECM-receptor interaction, protein digestion and absorption, and cytokine-cytokine receptor interaction. Additionally, compared with SE, 204 TFs(including FOS, EGR1, POU5F1, SOX2, and PAX6)were up-regulated, and 80 TFs(including HAND2, HOXB6, HOXB5, HOXA5, and HOXB8)were down-regulated in OSE cells. Furthermore, we identified 6 up-regulated and 9 down-regulated hub genes in OSE cells, and constructed TF-gene and TF-miRNA regulatory networks based on these hub genes.CONCLUSIONS:The transcriptome characteristics of OSE and SE cells were elucidated through RNA-seq analysis. These findings may provide a novel insight for studies on the development and in vitro directed induction of OSE and corneal epithelial cells.

Objective:To compare the effects of different test meals on postprandial triglycerides and to optimize the standard meal composition and the blood sampling protocol for the oral fat tolerance test.Methods:This study is a prospective, open-label, randomized, cross-over trial. In March 2023, 36 volunteers were recruited in Hebei General Hospital. They underwent a health examination and oral glucose tolerance test. Twenty-six healthy volunteers(11 males and 15 females) were included in this study, with an average age of(39.08±4.56) years. Each volunteer received 75 g protein meal, 75 g fat meal, 700 kcal fixed-calorie high-fat mixed meal, and a high-fat mixed meal with energy adjusted based on 10 kcal/kg body weight. A one-week washout period of regular diet was applied before each trial. Blood was collected at fasting status and 1, 2, 3, 4, 5, and 6 hours after a meal to detect serum triglycerides, total cholesterol, low density lipoprotein-cholesterol(LDL-C), high density lipoprotein-cholesterol(HDL-C), glucose, and insulin. The variations of postprandial metabolic indicators over time following the consumption of different test meals were analyzed. The disparities in postprandial metabolic responses between the two types of mixed meals were compared.Results:The protein meal, fat meal, fixed-calorie high-fat mixed meal, and adjusted-calorie high-fat mixed meal resulted in postprandial triglyceride increases of 22.45%, 115.40%, 77.14%, and 63.63%, and insulin increase of 560.43%, 85.69%, 554.18%, and 598.97%, respectively, and with reductions in total cholesterol, LDL-C, and HDL-C ranging from 5.64%-21.81%, respectively. The blood glucose changed slightly. Changes in metabolic indicators mainly occured within 4 hours. The comparison of the characteristics of postprandial triglycerides between the two high-fat mixed meals showed no statistically significant differences( P>0.05). Conclusion:A standardize protocol with a 700 kcal fixed-calorie high-fat mixed meal as test meal, and blood lipid levels measured at fasting and at 1, 2, 3, and 4 hours after consumption, can serve as an optimized approach for oral fat tolerance test.

Ten novel podophyllotoxin derivatives (IIIa-IIIi and IV) were synthesized by three-step reactions using podophyllotoxin and N-benzyloxycarbonyl glycine-L-proline as raw material. The structures of the target compounds were confirmed by 1H NMR, 13C NMR and MS. MTT method was used to test anti-tumor activity of the target compounds on HepG2, THP-1, HeLa and MCF-7 cells. The results showed that all the target compounds had inhibitory activity against HepG2, THP-1, HeLa and MCF-7 cells, and the inhibitory activity of IIIa on HepG2 cells was the most prominent with an IC50 value of 0.58 nmol/L. The binding mode of compound IIIa and FAPα was studied by molecular docking. Compound IIIa could bind to multiple sites of FAPα enzyme.

ObjectiveTo observe the effects of transcranial direct current stimulation (tDCS) on insomnia, anxiety and depression in patients with post-sroke insomnia (PSI). MethodsFrom January, 2020 to May, 2021, 44 patients with PSI from Beijing Bo'ai Hospital were randomly divided into control group (n = 22) and experimental group (n = 22). On the basis of conventional treatment, the experimental group accepted tDCS, and the control group accepted sham stimulation for four weeks. Pittsburgh Sleep Quality Index (PSQI) and sleep monitoring system based on cardiopulmonary coupling technology were used to evaluate the sleep quality of the patients before and after treatment. Hamilton Anxiety Scale (HAMA) and Hamilton Depression Scale (HAMD) were used to evaluate mood. ResultsTwo cases dropped out in each group. After treatment, the scores of PSQI, HAMA and HAMD decreased in both groups (t > 8.575, P < 0.001), and the scores of PSQI and HAMA were better in the experimental group than in the control group (t > 2.811, P < 0.01), however, there was no significant difference in the scores of HAMD between two groups (t = 1.756, P > 0.05). After treatment, the sleep quality index, total sleep time, sleep latency, sleep efficiency and wake conversion times improved (|t| > 4.721, P < 0.001), and the rapid eye movement time prolonged in the experimental group (t = -2.851, P = 0.010); the sleep quality index, total sleep time, sleep efficiency and wake conversion times were better in the experimental group than in the control group (t > 2.190, P < 0.05), however, no significant difference was found in the sleep latency and rapid eye movement time between two groups (|t| < 1.073, P > 0.05). ConclusiontDCS could improve the sleep quality and anxiety in PSI patients, and has little effect on sleep structure.

Objective : To investigate the effect of the combination of stem cell leukemia(SCL)recombinant lentivirus and connexin 43 ( Cx43 ) recombinant lentivirus on the function of diabetic cystipathy ( DCP) in guinea pigs. Methods : After 90 healthy guinea pigs were fed normally for 1 week, streptozotocin(STZ)was injected intraperito⁃ neally at 200 mg/kg in a single dose, and random blood glucose was monitored weekly. After 12 weeks of normal feeding, 40 guinea pigs meeting the criteria were screened by urodynamic examination and randomly divided into 4 groups: diabetic group, SCL group, Cx43 group, and SCL + Cx43 group. In the diabetic group, 0. 2 ml of empty lentivirus without gene was instilled into the bladder through the urethra, in the SCL and Cx43 groups, 0. 2 ml of SCL lentivirus and Cx43 lentivirus were instilled using the same method, in the SCL + Cx43 lentivirus group, 0. 2 ml of each SCL and Cx43 lentivirus was instilled through the urethra. After 14 days of transfection, urodynamic examination was performed, and then the guinea pigs were executed after the examination, and the bladders were quickly removed for frozen bladder sections and fluorescent double staining. Results : There were no differences in urodynamic examinations between the SCL and Cx43 groups compared to the diabetic group( P > 0. 05 ) . Urodynamic examination in the SCL + Cx43 group showed improvement in detrusor pressure, abdominal pressure and bladder pressure compared to the diabetic group(P < 0. 05) . In laser confocal experiments, the number of interstitial cells of Cajal (ICC) was reduced in the diabetic group, and the spindle structure and cell protrusions were obviously destroyed and appeared in a state of cell lysis. In the SCL and Cx43 groups, there was an improvement in the spindle structure and cell protrusion of ICC⁃like cells, and there was no significant change in the number of cells. In the SCL + Cx43 group, there was an increase in the number of ICC⁃like cells, a significant improvement in the spindle structure and cell protrusion, and the formation of ICC⁃dimers. Conclusion Transurethral co⁃infusion of SCL and Cx43 gene recombinant lentivirus can be successfully transfected in guinea pig DCP bladder, which can restore the number and structure of damaged ICC cells and form ICC dimer structure, improve the pressure of diabetic bladder forced urinary muscle, improve the weakness of urination, ventral pressure voiding and other characteristics. It provides a new direction for the treatment of DCP.

Body donation is the main source of human specimens preparation in medical universities and colleges. The low donation rate in China restricts the improvement of basic medical research and clinical treatment seriously. In the investigation, deep-rooted traditional concepts, unsound legal system, and inadequate government propaganda have become the major factors influencing body donation. Combined with the practice of body donation in the localities, this study puts forward some measures to promote the body donation, such as giving targeted publicity, promoting the concept of civilized funeral and burial, strengthening legislation, and simplifying the body donation process, which would lay the foundation for the development of medical education and research.

Objective To explore the effects of Yiqi Jianpi decoction on inflammatory reaction, immune function and nutritional status in patients after surgery of laparoscopic rectal cancer. Methods Eighty-two patients with post-operative laparoscopic rectal cancer resection admitted to Pingxiang People's Hospital from June 2017 to March 2019 were enrolled, and according to whether traditional Chinese medicine (TCM) was used or not, they were divided into a western medicine treatment control group (control group) and a combined traditional Chinese and western medicine treatment group (combined group), 41 cases in each group. The control group was treated with symptomatic conventional western therapy, while the combined group was additionally treated with Yiqi Jianpi decoction based on the conventional western treatment as in the control group; the composition of the decoction: hedyotis herba, herba agrimoniae, radix astragali each 30 g; radix codonopsis, ganoderma lucidum each 20 g; rizoma atractylodes, poria cocos and prunella chinensis 15 g each; pinellia ternata 9 g, licorice root 6 g; all the above ingredients were together boiled with water and the fluid (decoction) was filtered, 150 mL once orally taken, twice a day, 21 days constituting one therapeutic course, and the clinical efficacy was evaluated after 1 course of therapy. The changes of TCM syndrome score, simple health situation scale (SF-36) score, inflammatory indexes, immune function and nutritional status of the two groups were observed before and after treatment. Results After treatment, the levels of TCM syndrome score, C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and CD8+ in the two groups were significantly lower than those before treatment, while the levels of SF-36 score, CD4+, CD4+/CD8+, transferrin (TRF) and albumin (Alb) were significantly higher than those before treatment (all P < 0.05), and the degree of changes in the combined group were more significant than those in control group [TCM syndrome score: 3.79±2.22 vs. 6.86±2.02, SF-36 score:75.18±4.13 vs. 64.17±4.01, CRP (mg/L): 4.54±1.31 vs. 5.71±2.21, IL-6 (μg/L): 20.21±2.21 vs. 26.12±2.13, TNF-α (μg/L): 33.04±4.56 vs. 36.41±3.23, CD4+: 0.37±0.03 vs. 0.35±0.03, CD8+: 0.25±0.03 vs. 0.28±0.03, CD4+/CD8+: 1.51±0.39 vs. 1.19±0.37, TRF (g/L): 3.05±0.41 vs. 2.28±0.42, Alb (g/L): 43.88±2.28 vs. 39.86±2.03, all P < 0.05]. Conclusion Yiqi Jianpi decoction in the treatment of patients after surgery of laparoscopic rectal cancer can effectively promote the recovery of gastrointestinal function, reduce the inflammatory reaction, elevate the immune function and improve clinical prognosis.

To construct a eukaryotic expression plasmid containing the luciferase reporter gene (Fluc) to quickly detect apoptosis. Four amino acids, Asp-Glu-Val-Asp (DEVD), the recognize motif of Caspase-3, were introduced into the middle of the Fluc-C and N fragment. Meanwhile, four amino acids, Asp-Glu-Val-Gly (DEVG), were selected as a negative control. Subsequently, the recombinant gene was cloned into the N and C terminal end of the split intein, and named as pFluc-DEVD and pFluc-DEVG. Then the plasmids were transfected into cells and renilla luciferase was co-transfected in each sample as an internal control for transfection efficiency. Then the apoptosis level was detected by the double luciferase reporter gene and the Western blotting analysis. The results showed that when apoptosis occurred, the content of firefly luciferase expressed in the pFluc-DEVD plasmid transfected group was about 3 times higher than pFluc-DEVG plasmid transfected group. Furthermore, Western blotting detection indicated that the Fluc level was significantly increased in pFluc-DEVD transfected group when pre-treated by apoptosis stimulants. The activation degree of Caspase-3 was closely related to the expression of Fluc, and had a significant statistical difference. These results confirmed that firefly luciferase protein expressed by pFluc-DEVD plasmid can be cleaved by the intracellular Caspase-3 enzyme, and this plasmid can accurately reflect the cell apoptosis level, which provides a useful method for quantitative detection of apoptosis.
Apoptosis ; Genes, Reporter ; Luciferases, Firefly ; Transfection

Objective To investigate he intervention effect of knockdown of activating transcription factor 4(ATF-4)on fructose-induced lipid accumulation in liver cells. Methods HepG2 cells were divided into the control group(C),high fructose group(F),high-fructose+negative control group(F+NC)and high-fructose+ATF-4 siRNA group(F+ATF-4-). The mRNA level of gens of the upstream transcriptional factors and ERS markers was detected. The protein level of ACC,FAS and SCD-1 was also detected. Results Compared with group C,the mRNA expression of SREBP-1c,ChREBP,GRP78 and CHOP was increased(P < 0.01),while ATF-4 knock-down decreased the expression of the above genes(P < 0.01 ,respectively). Compared with group C ,the protein expression of ACC,FAS and SCD-1 was increased in group F(P<0.01,respectively). While ATF-4 knockdown decreased the protein expression of ACC ,FAS and SCD-1. Conclusions ATF-4 knockdown can improve the lipid steatosis induced by fructose through down-regulating lipid lipogenesis ,indicating ATF-4 possesses a regulatory effect on lipogenesis.

Objective To observe the effects of transcranial direct current stimulation (tDCS) on depression, anxiety and cognitive function in patients with post-stroke depression (PSD). Methods From February to August, 2017, 26 patients with PSD were randomly divided into control group (n=13) and treatment group (n=13). All the patients accepted sertraline, while the treatment group accepted tDCS, and the control group accepted sham stimulation. They were evaluated with Hamilton Depression Scale (HAMD), Hamilton Anxiety Scale (HAMA) and Mini-Mental State Examination (MMSE) before and after treatment. Results Twelve patients in the treatment group and ten patients in the control group completed the trial. The scores of HAMD, HAMA and MMSE improved in both groups (t>2.331, P<0.05), and there was no significant difference between two groups (t<1.642, P>0.05). The scores of anxiety/somatization factor, cognitive impairment factor, retardation factor and despair factor of HAMD improved in the treatment group after treatment (t>2.493, P<0.05), while only despair factor improved in the control group (t=2.862, P<0.05). The score of retardation factor was significantly different between two groups (t=2.354, P<0.05).Conclusion tDCS may work for the depression in PSD patients.