Objective In this study,we analyzed the transcriptome sequences of Kupffer cells exposed to simulated microgravity for 3 d and conducted biological experiments to determine how microgravity initiates apoptosis in Kupffer cells. Methods Rotary cell culture system was used to construct a simulated microgravity model.GO and KEGG analyses were conducted using the DAVID database.GSEA was performed using the R language.The STRING database was used to conduct PPI analysis.qPCR was used to measure the IL1B,TNFA,CASP3,CASP9,and BCL2L11 mRNA expressions.Western Blotting was performed to detect the level of proteins CASP3 and CASP 9.Flow cytometry was used to detect apoptosis and mitochondrial membrane cells.Transmission electron microscopy was used to detect changes in the ultrastructure of Kupffer cells. Results Transcriptome Sequencing indicated that simulated microgravity affected apoptosis and the inflammatory state of Kupffer cells.Simulated microgravity improved the CASP3,CASP9,and BCL2L11 expressions in Kupffer cells.Annexin-V/PI and JC-1 assays showed that simulated microgravity promoted apoptosis in Kupffer cells.Simulated microgravity causes M1 polarization in Kupffer cells. Conclusion Our study found that simulated microgravity facilitated the apoptosis of Kupffer cells through the mitochondrial pathway and activated Kupffer cells into M1 polarization,which can secrete TNFA to promote apoptosis.

Objective:To investigate the association between tumor necrosis factor-β (TNF-β) gene polymorphisms and genetic predisposition to gastric cancer, and to analyze the relationship between specific genotype of TNF-β and serum levels of TNF-β.Methods:Using case control study, we selected 153 patients with gastric cancer in Fudan University Shanghai Cancer Center between September 2021 and December 2022 as the gastric cancer group, and 150 healthy individuals were chosen as the healthy control group. In the previous study, 30 peripheral blood DNA samples of gastric cancer patients and healthy controls respectively were amplified by conventional PCR, which were sequenced to identify the genotype frequencies of TNF-β polymorphic loci (rs1041981, rs2229092, rs2229094 and rs78613290); consequently, Allele-Specific Quantitative PCR was used to further detect and analyze the genotype and genotype frequencies of TNF-β polymorphic loci; serum TNF-β levels were measured by Enzyme-Linked Immunosorbent Assay (ELISA), and the relationship with specific genotypes of TNF-β was analyzed. Chi-square test and Fisher test were used to analyze the genotype distribution frequency of TNF-β polymorphic loci, and non-parametric statistics was used to analyze the differences in serum TNF-β expression levels.Results:The sequencing results showed that the genotype distribution of rs1041981 in gastric cancer group was CC 16.67% (5/30), CA 40.00% (12/30) and AA 43.33% (13/30). The genotype distribution in control group was CC 40.0% (12/30), CA 43.33% (13/30), AA 16.7% (5/30). The difference of genotype frequency between the two groups was statistically significant (χ 2=6.478, P=0.039). The genotypes of the polymorphic loci rs2229092 in both groups were AA, AG, and GG, with no statistically significant difference between the two groups (χ 2=1.888, P=0.612). The distribution frequencies of the genotypes of the polymorphic loci rs2229094 (TT and TC) and rs78613290 (GG and AG) showed no statistically significant differences between the two groups (both P>0.05). Further validation with an expanded clinical samples (153 cases in the gastric cancer group and 150 cases in the control group) found that the difference of rs1041981 genotype distribution between the gastric cancer group [CC 15.69%(24/153), CA 54.9%(84/153), AA 29.4%(45/153)] and the control group [CC 27.3%(41/150), CA 58.0%(87/150), AA 14.7%(22/150)] was significantly different (χ 2=12.366, P=0.002). Analysis of the influence of different allele frequencies on the risk of gastric cancer revealed that the odds ratio ( OR) of the A allele of rs1041981 for the risk of gastric cancer compared to the C allele was 1.701 (95% CI 1.235?2.355). Gene phenotype analysis combining the clinicopathological characteristics of gastric cancer patients found that the distribution frequency of the rs1041981 genotype was significantly different among groups of different genders, tumor invasion depths, and the lymph node metastasis, with statistically significant differences (All P>0.05). Additionally, gastric cancer patients with rs1041981 AA genotypes had higher serum TNF-β expression levels than those with CA and CC genotypes, (both P<0.05). Conclusions:The gene type frequency of the TNF-β gene polymorphic loci (rs1041981, C>A) exhibited significant differences between the gastric cancer group and the healthy control group. The presence of the A allele in rs1041981 site increased the susceptibility to gastric cancer, and patients with different gene types displayed vaning levels of serum TNF-β, among which AA genotype ranks the highest level.

Purpose: Patients with advanced biliary tract cancer (BTC) have a poor survival. We aim to evaluate the efficacy and safety of nab-paclitaxel plus gemcitabine and cisplatin regimen in Chinese advanced BTC patients. Materials and Methods: Eligible patients with locally advanced or metastatic BTC administrated intravenous 100 mg/m2 nab-paclitaxel, 800 mg/m2 gemcitabine, and 25 mg/m2 cisplatin every 3 weeks. The primary endpoint was progression-free survival (PFS). The secondary endpoints included overall survival (OS) and adverse events, while exploratory endpoint was the association of biomarkers with efficacy. Results: After the median follow-up of 25.0 months, the median PFS and OS of 34 enrolled patients were 7.1 months (95% confidence interval [CI], 5.4 to 13.7) and 16.4 months (95% CI, 10.9 to 23.6), respectively. The most common treatment-related adverse events at ≥ 3 grade were neutropenia (26.5%) and leukopenia (26.5%). Survival analyses demonstrated that carcinoembryonic antigen (CEA) levels could monitor patients’ survival outcomes. A significant increase in the number of infiltrating CD4+ cells (p=0.008) and a decrease in programmed death-1–positive (PD-1+) cells (p=0.032) were observed in the response patients. Conclusion In advanced BTC patients, nab-paclitaxel plus gemcitabine and cisplatin regimen showed therapeutic potential. Potential prognostic factors of CEA levels, number of CD4+ cells and PD-1+ cells may help us maximize the efficacy benefit.

Objective:The investigation and research on the application status of Hepatic Venous Pressure Gradient (HVPG) is very important to understand the real situation and future development of this technology in China.Methods:This study comprehensively investigated the basic situation of HVPG technology in China, including hospital distribution, hospital level, annual number of cases, catheters used, average cost, indications and existing problems.Results:According to the survey, there were 70 hospitals in China carrying out HVPG technology in 2021, distributed in 28 provinces (autonomous regions and municipalities directly under the central Government). A total of 4 398 cases of HVPG were performed in all the surveyed hospitals in 2021, of which 2 291 cases (52.1%) were tested by HVPG alone. The average cost of HVPG detection was (5 617.2±2 079.4) yuan. 96.3% of the teams completed HVPG detection with balloon method, and most of the teams used thrombectomy balloon catheter (80.3%).Conclusion:Through this investigation, the status of domestic clinical application of HVPG has been clarified, and it has been confirmed that many domestic medical institutions have mastered this technology, but it still needs to continue to promote and popularize HVPG technology in the future.

Objective:To investigate the role and regulatory mechanism of UGT2A3 differential expression in colorectal carcinogenesis.Methods:Nine CRC datasets were downloaded from GEO database and TCGA. R language was used to analyze the differential expression of UGT2A3 in cancer and normal tissues. According to the expression level of UGT2A3 in TCGA, the top 20 samples with the highest expression and the lowest expression were selected from normal tissues and CRC tissues, respectively. The abundance of immune cells and immune enrichment score were compared, the differentially expressed genes and differentially expressed miRNAs were screened, and the pathway enrichment analysis of differentially expressed genes was performed.Results:UGT2A3 was down regulated in all 9 CRC datasets. In all sample types, compared with the UGT2A3 high expression group, the UGT2A3 low expression group had significantly higher ImmuneScore, EstimateScoreandStromalScore, and had higher abundance of immune cells (except memory B cells). In normal tissues, the differential expression of UGT2A3 mainly affects cancer-related pathways, while in tumor tissues, it mainly affects metabolic pathways. miR-194-2, miR-224 and miR-551b were differentially expressed in all groups, which were considered as potential UGT2A3 upstream regulatory genes in CRC.Conclusions:UGT2A3, miR-194-2, miR-224 and miR-551b can be used as potential biomarkers for the diagnosis of CRC.

Objective:To explore the clinical prognosis of early endarteritis (within 2 weeks) and late endarteritis (after 2 weeks) after renal transplantation.Methods:A total of 81 cases with higher creatinineand receiving renal biopsy after renal transplantation were recruited from September 2001 to December 2014. They were divided into early endarteritis group (n=43) and late endarteritis group (n=38). Baseline profiles, serum creatine, glomerular filtration rate (GFR) before and after treatment, steroid resistance, reversal rate, graft loss and survival rate were analyzed for two groups.Results:Early endarteritis group showed worse serum creatine and GFR than late endarteritis group before rejection. Early endarteritis group had a higher rate of treatment with steroid plus antibody (86 %) than that of late endarteritis group (86 %vs.18.6 %, P<0.05). No significant inter-group difference existed in graft loss (23.3 % vs.10.5 %, P=0.131). The survival curve of transplanted kidney showed no significant inter-group difference insurvival time. Conclusions:The status of patients with early simple endothelitis is significantly worse than that of those with late simple endothelitis. However, after active treatments, the prognosis of patients with early simple endothelitis is not inferior to that of those with late simple endothelitis.

Objective:To investigate the thermal injury effects on human HaCaT cells under simulated microgravity environment.Methods:The human HaCaT cells were collected and divided into simulated microgravity thermal injury (SMGTI) group, normal gravity thermal injury (NGTI) group, and normal gravity false injury (NGFI) group according to the random number table. Cells in NGTI and NGFI groups were cultured routinely in culture bottle, and cells in SMGTI group were cultured in the rotary cell culture system to simulate microgravity environment. Cells in SMGTI and NGTI groups were bathed in hot water of 45 ℃ for 10 minutes to make thermal injury model, and cells in NGFI group were bathed in warm water of 37 ℃ for 10 minutes to simulate thermal injury. At post injury hour (PIH) 12, cell morphology of 3 groups was observed under inverted phase contrast electron microscope. At PIH 2, 6, and 12, single cell suspension in the 3 groups was collected to detect the cell cycle by flow cytometer and the mRNA expressions of heat shock protein 70 (HSP70), matrix metalloproteinase 9 (MMP-9), and cysteine-aspartic protease 3 (caspase-3) by real time fluorescence quantitative reverse transcription polymerase chain reaction, and the experiments were repeated for 3 times. At PIH 2, 6, and 12, cell culture supernatant in the 3 groups was collected to detect the concentration of heparin-binding epidermal growth factor (HB-EGF) by enzyme linked immunosorbent assay method, the experiment was repeated for 3 times. The sample in each group and each time point was 3. Data were statistically analyzed with analysis of variance for factorial design, one-way analysis of variance, least significant difference test, Kruskal-Wallis H test, and Mann-Whitney U test. Results:(1) At PIH 12, cells in NGFI group showed regular shape and regular arrangement, with no cell debris. The cell shape in NGTI group was generally regular, with fewer cell debris and closer arrangement than that in NGFI group. The cells in SMGTI group showed more irregular shapes, different sizes, and dead cell debris. (2) The percentage of G1 phase cells in NGTI group was significantly higher than that in NGFI group and SMGTI group at PIH 2, respectively ( P<0.05), and the percentage of G1 phase cells in NGTI group was significantly lower than that in NGFI group and SMGTI group at PIH 6 and 12, respectively ( P<0.05). The percentage of G2/M phase cells in NGTI group was significantly lower than that in SMGTI group at PIH 2 ( P<0.05), and the percentage of G2/M phase cells in NGTI group was significantly higher than that in NGFI group and SMGTI group at PIH 6 and 12, respectively ( P<0.05). The percentage of S phase cells in NGTI group at PIH 2, 6, and 12 was significantly higher than that in SMGTI group ( P<0.05), and the percentage of S phase cells in NGTI group at PIH 2 and 6 was significantly lower than that in NGFI group ( P<0.05). (3) The HSP70 mRNA expressions of cells in NGTI group were 2.50±0.30 and 3.99±0.35 at PIH 2 and 6, which were significantly higher than 1.14±0.15 and 0.82±0.27 in NGFI group ( P<0.05), and 1.17±0.53 and 1.65±0.59 in SMGTI group ( P<0.05). The MMP-9 mRNA expression of cells in SMGTI group was significantly higher than that in NGTI group at PIH 2, 6, and 12, respectively ( Z=-2.319, -2.882, -2.908, P<0.05). At each time point after injury, the mRNA expression of caspase-3 of cells in NGTI group was similar to that in NGFI group and SMGTI group, respectively ( P>0.05). (4) The concentration of HB-EGF in cell culture supernatant of NGTI group was significantly lower than that in NGFI group at PIH 2, 6 and 12 ( P<0.05), and the concentration of HB-EGF in cell culture supernatant of SMGTI group was significantly higher than that in NGTI group at PIH 2 and 6 ( P<0.05). Conclusions:The proliferation and secretion functions and expression of wound repair related protein of human HaCaT cells inflicted with thermal injury in simulated microgravity environment showed complex and diversified changes, which provide theoretical basis for further research on damage repair under weightlessness.

Objective To investigate the effects of simulated microgravity by RCCS on proliferation and cell cytoskeleton of human HaCaT keratinocyte. Methods The rotary cell culture system (RCCS) was used to simulate the microgravity environment, and human HaCaT keratinocytes were divided randomly(random number) into the simulated microgravity group (SMG) and normal gravity group (NG). HaCaT cells in the two groups were harvested respectively after 32, 36 and 42 h culture. The HaCaT cells proliferation and cycles were detected by flow cytometry, the concentration of hb-EGF in supernatant was detected by ELISA, and the cell cytoskeleton was observed after 42 hours' culture under laser confocal microscope with FITC-labeled technique. SPSS 23.0 statistical software was used for statistical analysis, and P <0.05 was considered statistically significant. Results The flow cytometry showed that the proportions of human HaCaT keratinocytes in G1 and G2/M phases were increased while the proportion of HaCaT cells in S stage was decreased significantly after 32, 36 and 42 h RCCSculture compared with those in the normal gravity group. The HaCaT cells in G1 stage were declined along with incubation time. ELISA results showed that the hb-EGF concentration in HaCaT supernatant under simulated microgravity culture for 24 and 36 h was lower than that in the normal control group (P<0.01). The laser confocal microscope revealed that the HaCaT fluorescence intensity was decreased,and there were disordered microfilaments, structural ambiguity, pseudopodia reduction and irregularshape among FITC-labeled HaCaT cells cultured 42 h in RSSC compared with the normal gravity group.Conclusions RCCS simulated microgravity environment could inhibit the cell cycle transformation and proliferation of human HaCaT keratinocyte, affect the keratinocyte-secreting function, and induce alterations of the cell cytoskeleton.

The rotary cell culture system(RCCS)was used to simulate the microgravity environment, and FRTL-5 cells were divided into simulated microgravity group(SMG)and normal gravity group(NG). FRTL-5 cells were harvested after treatment for 6, 12, 24, and 36 h, the cell viability was measured by MTT assay, and the cells cycles were detected by flow cytometry. The ultrastructure of FRTL-5 cells was observed under laser confocal microscope with FITC-labeled technique. The MTT assay showed that the proliferation of FRTL-5 cells was significantly inhibited after RCCS treatment for 6, 12, 24, and 36h compared with NG(P<0.05), in which the most obvious effect was observed at 24h. The flow cytometry showed that the proportion of FRTL-5 cells at G1 stage in RCCS group was increased significantly after 6, 12, 24, and 36h compared with NG(P<0.05), while the proportion of FRTL-5 cells at S stage was decreased significantly(P<0.05)except that cultured with RCCS for 6 h. The proportion of FRTL-5 cells at G2/M stage was decreased in early phase(6-12 hours)of RCCS culture, with the lowest at 12h and transient increase at 24h of RCCS culture. The laser confocal microscope revealed that there were local microfilament depolymerization, tension fibers decrease, structure disorder, cellular pseudopodia reduction, and irregular shape among FITC-labeled FRTL-5 cells cultured with RCCS for 36h. (Chin J Endocrinol Metab, 2018, 34: 598-601)

Objective To deepen and promote the development of nursing higher education internationalization through the analyzation of the status,existing problems and future trends of research about nursing higher education.Methods Searching literatures in China National Knowledge Infrastructure (CNKI),VIP and WANFANG databases from 1984 until 2014.Results 79 references were selected from 44 journals and the numbers increased each year.69.62％ (55/79) of them were accepted by statistical source core journals and 54.43％ (43/79) of them were funded.The authors came from 19 provinces.There were only 4 articles reported international education about graduates and most of them were about undergraduate students.The main reformation measures included curriculum and english teaching reform et al.Conclusions The educator paid more attention to nursing higher education internationalization than before,also the policy's support and the quality of literatures were optimistic.However,the breadth,depth and integrity of research should be strengthened in future.