BACKGROUND:There are many kinds of cell media with different components,which have a great influence on cell growth.Several studies in and outside China have used serum-free media containing fetal bovine serum for in vitro amplification culture,but the use of media containing human peripheral blood serum to directionally induce human umbilical cord mesenchymal stem cells to neural stem cells and human peripheral blood serum to promote neural stem cells to differentiate into other nerve cells,so far there are few relevant studies. OBJECTIVE:To observe the feasibility of inducing human umbilical cord mesenchymal stem cells cultured with human peripheral blood serum into neural stem cells. METHODS:(1)Human umbilical cord mesenchymal stem cells were cultured in DMEM/F-12 culture medium containing 10%human peripheral blood serum by volume fraction.Flow cytometry analysis was performed at the third passage to identify surface markers and alizarin red staining was used to detect osteogenic differentiation function.(2)The third-generation human umbilical cord mesenchymal stem cells were induced into neural stem cells using DMEM/F-12 medium containing 0.5%N2,1.5%B27,20 ng/mL basic fibroblast growth factor,and 20 ng/mL epidermal growth factor,and their surface markers were identified.(3)Well-growing human umbilical cord mesenchymal stem cell-derived neural stem cells were taken to prepare a single cell suspension.They were evenly inoculated into 96-well plates and incubated with DMEM/F-12 culture medium containing 10%human peripheral blood serum for 8 days.Then,hematoxylin-eosin staining,microtubule-associated protein 2,and glial fibrillary acidic protein immunofluorescence staining were performed to detect the differentiation of neural stem cells into other neural cells. RESULTS AND CONCLUSION:(1)Human umbilical cord mesenchymal stem cells cultured with human peripheral blood serum grew in a spiral-like pattern and were distributed in multiple layers,with directional arrangement.The surface of human umbilical cord mesenchymal stem cells highly expressed CD44,CD105,CD29,and CD73.Cells stained with alizarin red showed a color reaction.(2)Human umbilical cord mesenchymal stem cells cultured with human peripheral blood serum could be induced to differentiate into neural stem cells,and the surface of neural stem cells was highly expressed with CD44,CD105,CD29,CD73,Nestin,NF-L,and GALC.(3)On day 8 of induced differentiation of neural stem cells,after staining with hematoxylin and eosin,it was found that the protruding protrusions were longer,with more branches and adjacent cells connected,presenting typical neural cell morphology.Immunofluorescence staining for microtubule-associated protein 2 and glial fibrillary acidic protein was positive.It is concluded that human umbilical cord mesenchymal stem cells cultured by human peripheral blood serum can be directly induced to differentiate into neural stem cells.Under the action of human peripheral blood serum,neural stem cells can differentiate into other neural cells as the culture time prolongs.

Objective To study the effect of low concentrations of sodium fluoride on the osteogenic/odontogenic differentiation of human dental pulp cells(hDPCs)in vitro.Methods This study was reviewed and approved by the Ethics Committee.hDPCs were cultured using a modified tissue explant technique in vitro.The effects of different con-centrations of sodium fluoride on the proliferation of hDPCs were measured by methylthiazol tetrazolium(MTT)assay.Appropriate concentrations were added to the osteogenic/odontogenic differentiation induction medium,and the cells were induced in vitro.Alizarin red S staining was used to detect the osteoblastic/odontogenic differentiation ability of the cells,and the mRNA expression of the key differentiation factors was detected by RT-qPCR.Moreover,the expres-sion of key molecules of endoplasmic reticulum stress(ERS)was detected by RT-qPCR and Western blot.The data were analyzed with the SPSS 18.0 software package.Results Low concentration of NaF(0.1 mmol/L)could stimulate cell proliferation in vitro,while a high concentration(5-10 mmol/L)could inhibit cell proliferation(P＜0.05).According to the literature and the experimental data,0.1 mmol/L NaF was selected as the following experimental concentration.The levels of alizarin red S staining were increased after NaF induction of mixed osteogenic/odontogenic differentiation in vi-tro.The mRNA expression levels of key molecules for osteogenic/odontogenic differentiation,dentin sialophosphoprotein(DSPP),bone sialoprotein(BSP)and osteocalcin(OCN),were increased(P＜0.05).The mRNA levels of ERS markers(splicing x-box binding protein-1(sXBP1),glucose-regulated protein 78(GRP78)and activating transcription Factor 4(ATF4)were increased in NaF-treated cells.The protein expression levels of key ER stress molecules(phosphorylated RNA-activated protein kinase-like ER-resident kinase(p-PERK),phosphorylated eukaryotic initiation factor-2α(p-eIF2α)and ATF4)were higher in NaF-treated cells.Conclusion A low concentration of NaF promotes the osteogenic/odontogenic differentiation of hDPCs and increases the level of ER stress.

Objective To use bioinformatics analysis to identify the key genes and signaling pathways involved in cardiac dysfunction after acute ischemic stroke.Methods The GSE102558 dataset was downloaded from the Gene Expression Omnibus(GEO)database,and genes with values of P<0.05 and|log2FC|>0.6 were identified as being differentially expressed.The MCODE plugin in Cytoscape software performed a functional module analysis of the protein-protein interaction(PPI)network,while the CytoHubba plugin screened for core genes.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were also performed.A middle cerebral artery occlusion model was constructed to verify core gene expression using real-time PCR.Results Among the screened differential genes,385 were upregulated and 354 were downregulated.The top ten core genes were Col1a1,Col1a2,Col3a1,Fbn1,Postn,Col5a1,Mmp3,Eln,Acta2,and Timp3.The GO enrichment mainly involved the extracellular matrix,the collagen fiber tissue,vascular development,and protease binding.KEGG was mainly enriched in protein digestion and absorption,the relaxin pathway,advanced gly-cation end products-receptor for advanced glycation end products,and platelet activation.Real-time PCR verified that Col3a1and Postn expressions decreased in the heart tissue after acute ischemic stroke.Conclusion Col3a1and Postnexpressions may be closely associ-ated with cardiac dysfunction occurrence and development after acute ischemic stroke.

Objective:To examine the potential of combining the blood urea nitrogen(BUN)/albumin(ALB)ratio(BAR)with soluble growth stimulation expressed gene 2(sST2)as an early warning indicator for myocardial injury in elderly patients with sepsis in the emergency intensive care unit(EICU).Methods:The clinical data of elderly patients with sepsis admitted to the EICU of the Emergency Medicine Department at Harrison International Peace Hospital of Hebei Medical University from August 2018 to August 2022 were prospectively analyzed.The patients were divided into two groups based on the presence or absence of myocardial injury: the myocardial injury group and the non-myocardial injury group.The general clinical data and laboratory indexes of the two groups were compared, and the BAR was calculated.The correlation between BAR and myocardial injury in elderly patients with sepsis was analyzed.Binary Logistic regression analysis was used to identify the risk factors for myocardial injury in elderly patients with sepsis in the EICU.MedCalc software was employed to determine the early warning value of the combined sST2 and BAR for myocardial injury in elderly patients with sepsis in the EICU.Results:A total of 165 cases were analyzed, with 106 cases(64.24%)showing myocardial injury.It was found that elderly sepsis patients with lung and abdominal infection were more likely to experience myocardial injury( P<0.05 for all).In comparison to the group without myocardial injury, the levels of inflammatory markers such as white blood cell count(WBC), neutrophil to lymphocyte ratio(NLR), lactic acid, and procalcitonin(PCT), as well as combined markers BAR and sST2, were higher in elderly sepsis patients with myocardial injury upon admission.Correlation analysis results revealed significant positive correlations between BAR and lactic acid, PCT, and C-reactive protein(CRP)within 24 hours of admission to EICU in elderly sepsis patients with myocardial injury.Among these correlations, the strongest was observed between BAR and PCT( r=0.417, P<0.001).Additionally, BAR exhibited a positive correlation with acute physiology and chronic health evaluation scoring system(APACHEⅡ)scores( r=0.241, P=0.002).Furthermore, BAR showed positive correlations with myocardial injury markers sST2 and cTnI( r=0.327, 0.307, P<0.05 for all).Logistic regression analysis revealed that septic shock( OR=2.406, P=0.008), decreased left ventricular ejection fraction(EF)( OR=0.939, P=0.015), BAR( OR=2.205, P=0.044), lactic acid( OR=1.137, P=0.014), and sST2 elevation( OR=1.016, P=0.020)were identified as independent risk factors for predicting myocardial injury in elderly patients with sepsis.The results of ROC analysis indicated that BAR had a high early warning value for the occurrence of myocardial injury in elderly patients with EICU sepsis[area under curve(AUC)0.651, P<0.05], with an optimal cut-off value of 0.32(sensitivity 77.4%, specificity 60.2%).Furthermore, the combined detection of BAR and sST2 demonstrated a higher early warning value for the occurrence of myocardial injury in elderly patients with sepsis(AUC 0.697, P<0.05).The mortality rate of patients with myocardial injury below a cut-off value of 0.32 was 36.00%(9/25), while the mortality rate of patients with myocardial injury equal to or above 0.32 was 66.67%(54/81).The difference between the two groups was statistically significant( χ2=8.624, P=0.003). Conclusions:Both BAR and sST2 are considered independent risk factors for myocardial injury in elderly patients with sepsis.The combined detection of BAR and sST2 provides a more accurate prediction for the occurrence of myocardial injury in these patients.

Objective: To investigate the risk factors for yersiniosis, so as to provide insights into prevention of yersiniosis. Methods: The patients with yersiniosis admitted to the clinics in the surveillance site of Chengbei Township of Jin'an District and Chengnan Township of Yu'an District in Lu'an City from 2013 to 2021 were included as the case group, and the healthy family members matched to cases were selected as the family control group, while normal residents with a 1︰2 match in the same village, gender, and age difference within 5 years were included in the community control group. Participants' demographics, hand-washing and eating habits, living environment hygiene, poultry and livestock feeding were collected using questionnaire surveys, and factors affecting yersiniosis were identified using a multivariable conditional logistic regression model. Results: There were 43 cases in the case group, with a median (interquartile range) age of 45 (34) years, 91 cases in the family control group, with a median (interquartile range) age of 36 (36) years and 86 cases in the community control group, with a median (interquartile range) age of 46 (34) years. Multivariable conditional logistic regression analysis showed that compared with the family control group, the habit of drinking unboiled water (OR=6.721, 95%CI: 1.765-25.588), and direct consumption of food stored in the refrigerator (OR=7.089, 95%CI: 1.873-26.829) were risk factors for yersiniosis in the case group; and compared with the community control group, not washing hands after contacting with poultry and livestock (OR=50.592, 95%CI: 2.758-927.997), habit of eating raw vegetables and fruits (OR=5.340, 95%CI: 1.022-27.887), direct consumption of food stored in the refrigerator (OR=19.973, 95%CI: 2.118-188.336), and unclean refrigerator (OR=12.692, 95%CI: 1.992-80.869) were risk factors for yersiniosis in the case group. Compared with the family and community control groups, not washing hands after contacting with poultry and livestock (OR=4.075, 95%CI: 1.427-11.637), habit of drinking unboiled water (OR=4.153, 95%CI: 1.331-12.957), habit of eating raw vegetables and fruits (OR=4.744, 95%CI: 1.609-13.993), and direct consumption of food stored in the refrigerator (OR=5.051, 95%CI: 1.773-14.395) were risk factors for yersiniosis in the control group. Conclusion Unhealthy habits such as eating raw vegetables and fruits, drinking unboiled water, direct consumption of food stored in the refrigerator, unclean refrigerator, and not washing hands after contacting poultry and livestock may increase the risk of yersiniosis.

BACKGROUND: LY01005 (Goserelin acetate sustained-release microsphere injection) is a modified gonadotropin-releasing hormone (GnRH) agonist injected monthly. This phase III trial study aimed to evaluated the efficacy and safety of LY01005 in Chinese patients with prostate cancer. METHODS: We conducted a randomized controlled, open-label, non-inferiority trial across 49 sites in China. This study included 290 patients with prostate cancer who received either LY01005 or goserelin implants every 28 days for three injections. The primary efficacy endpoints were the percentage of patients with testosterone suppression ≤50 ng/dL at day 29 and the cumulative probability of testosterone ≤50 ng/dL from day 29 to 85. Non-inferiority was prespecified at a margin of -10%. Secondary endpoints included significant castration (≤20 ng/dL), testosterone surge within 72 h following repeated dosing, and changes in luteinizing hormone, follicle-stimulating hormone, and prostate specific antigen levels. RESULTS: On day 29, in the LY01005 and goserelin implant groups, testosterone concentrations fell below medical-castration levels in 99.3% (142/143) and 100% (140/140) of patients, respectively, with a difference of -0.7% (95% confidence interval [CI], -3.9% to 2.0%) between the two groups. The cumulative probabilities of maintaining castration from days 29 to 85 were 99.3% and 97.8%, respectively, with a between-group difference of 1.5% (95% CI, -1.3% to 4.4%). Both results met the criterion for non-inferiority. Secondary endpoints were similar between groups. Both treatments were well-tolerated. LY01005 was associated with fewer injection-site reactions than the goserelin implant (0% vs . 1.4% [2/145]). CONCLUSION: LY01005 is as effective as goserelin implants in reducing testosterone to castration levels, with a similar safety profile. TRIAL REGISTRATION ClinicalTrials.gov, NCT04563936.
Humans ; Male ; Antineoplastic Agents, Hormonal/therapeutic use* ; East Asian People ; Gonadotropin-Releasing Hormone/agonists* ; Goserelin/therapeutic use* ; Prostate-Specific Antigen ; Prostatic Neoplasms/drug therapy* ; Testosterone

Object:To explore surgical treatments for duodenal fistula with intra-abdominal infection.Methods:The data of 19 patients with duodenal fistula treated at the Affiliated Tumor Hospital of Zhenzhou University between Jan 2015 and Dec 2021 were analyzed retrospectively. Surgery is performed with duodenostomy or modified duodenal shunt procedures.Result:All patients were accompanied by intra-abdominal infection, including 9 duodenal stump fistulas. All patients successfully completed the operation,11cases underwent duodenostomy, 8 case underwent modified duodenal shunt procedures. operating time was 110(60-140)min, postoperative hospitalization time was 29(9-103)d. Two patients died postoperatively. Fistula heals in other patients.Conclusion:Surgical intervention for duodenal fistula should focus on controlling the source of infection, strengthening intestinal and abdominal drainage, and reducing postoperative complications.

Objective:To investigate the role of modification level of lysine trimethylation at position 27 of histone 3 (H3K27me3) on expression of anti-apoptotic protein B lymphocyte tumor-2 gene (BCL2) during arsenic-induced hepatocyte apoptosis.Methods:Rat liver BRL-3A cells were cultured in vitro. According to the arsenic treatment factor, the experiment was divided into two parts, in the first part arsenic was not added, the experiment was divided into normal, transfection reagent, negative transfection, H3K27me3 specific demethylase (JMJD3) small interfering RNA (siRNA) transfection and H3K27me3 methyltransferase (EZH2) siRNA transfection groups. In the second part arsenic was added, the experiment was divided into control, arsenic treatment, arsenic + negative transfection, arsenic + JMJD3siRNA transfection and arsenic + EZH2siRNA transfection groups. When arsenic was not added, the corresponding siRNA and transfection reagent was used to transfect cells at a ratio of 100 pmol : 7.5 μl for 6 h [the normal group was treated with phosphate buffer solution (PBS) of the same volume as transfection reagent], then the medium was changed and the cells were incubated for a total of 48 h. After 24 h of treatment with the above transfection and culture method in arsenic added group, a final concentration of 30 μmol/L sodium arsenite (NaAsO 2) was added and the cells were incubated for 24 h (the control group was treated with PBS with the same volume of NaAsO 2 for 24 h). Real-time cell analysis (RTCA) was used to measure the proliferation of BRL-3A cells in arsenic added group. Apoptosis of BRL-3A cells was analyzed by flow cytometry in arsenic added group. Western blotting was used to detect JMJD3, EZH2, H3K27me3 and BCL2 in no-arsenic and arsenic-added BRL-3A cells. The modification levels of H3K27me3 in BCL2 gene promoter regions were detected by chromatin immunoprecipitation of the cells exposed to arsenic. Results:There were statistically significant differences of the proliferation rates [control, arsenic treatment, arsenic + negative transfection, arsenic + JMJD3siRNA transfection and arsenic + EZH2siRNA transfection groups: (100.00 ± 10.43)%, (12.19 ± 3.37)%, (31.86 ± 1.95)%, (24.58 ± 3.64)%, (11.53 ± 1.11)%] and the apoptosis rates [(1.15 ± 0.04)%, (13.06 ± 1.33)%, (17.39 ± 0.22)%, (23.90 ± 1.66)%, (15.07 ± 0.88)%] between groups ( F = 146.50, 194.30, P < 0.001), correspondingly. The protein expression level of H3K27me3 in JMJD3siRNA transfection group was higher than that of normal, transfection reagent and negative transfection groups, while EZH2siRNA transfection group had an opposite result ( P < 0.05). The protein expression level of BCL2 in JMJD3siRNA transfection group was lower than that of normal, transfection reagent and negative transfection groups, while EZH2siRNA transfection group had an opposite result ( P < 0.05). The protein expression levels of H3K27me3 and BCL2 were not statistically significant differences between normal, transfection reagent and negative transfection groups ( P > 0.05). The protein expression levels of JMJD3, EZH2, H3K27me3 and BCL2 among control, arsenic treatment, arsenic + negative transfection, arsenic + JMJD3siRNA transfection and arsenic + EZH2siRNA transfection groups were compared, and the differences were statistically significant ( F = 26.56, 7.82, 9.81, 31.19, P < 0.05). Compared with control group, the protein expression levels of JMJD3 and EZH2 in arsenic treatment group were significantly reduced ( P < 0.05), and the protein expression level of H3K27me3 was higher ( P < 0.05), meanwhile the protein expression level of BCL2 was lower ( P < 0.05). Compared with arsenic + negative transfection group, the protein expression level of JMJD3 was significantly reduced in arsenic + JMJD3siRNA group, and the protein expression level of EZH2 was significantly reduced in arsenic + EZH2siRNA group ( P < 0.05). In addition, arsenic + JMJD3siRNA increased the level of H3K27me3 modification while reducing the protein expression of BCL2, while arsenic + EZH2siRNA had an opposite result ( P < 0.05). Compared with control group, the enrichment levels of H3K27me3 in BCL2 gene promoter regions (CHIP1 and CHIP2) in arsenic treatment group were significantly higher ( P < 0.05). Conclusion:Arsenic may inhibit the expression of BCL2 by increasing the enrichment level of H3K27me3 in the promoter regions of BCL2 gene, and promoting hepatocyte apoptosis.

Objective:To evaluate a nano-carbon lymphatic tracing method for patients with rectal cancer after neoadjuvant radiotherapy and chemotherapy .Method:Retrospective analysis was made on 88 patients of rectal cancer undergoing neoadjuvant chemoradiation at the Department of General Surgery, He′nan Cancer Hospital from Jan 2016 to May 2020.According to whether nano-carbon lymph node was used or not, patients were divided into nanocarbon tracer group (study group) and non-nanocarbon tracer group (control group).Results:There was statistically significant in the number of havested lymph nodes between the two groups [15(11-19) vs.9(5-12), Z=5.227, P<0.001], There was no statistically significant in the number of positive lymph nodes between the two groups [0(0-0.25) vs.0(0-1), Z=1.199, P=0.231]. There were significant differences in the ratio of patients with less than 7 lymph nodes(0/34 vs.18/54, χ 2=14.248, P<0.001) and patients with less than 10 lymph nodes (4/34 vs.29/54, χ 2=15.657, P<0.001). Conclusions:The injection of nanocarbon after neoadjuvant chemoradiotherapy can increase the number of harvested postoperative lymph nodes and the ratio of patients with lymph nodes ≥7 and ≥10, which is more beneficial for prediction of the prognosis of patients.

Objective:To investigate the effects of preoperative endoscopic mucosal injection of carbon nanoparticle tracer and intraoperative serosa injection of carbon nanoparticle tracer on the acquisition of lymph nodes in total gastrectomy for gastric cancer.Methods:The retrospective cohort study was conducted. The clinical data of 118 patients with gastric cancer who underwent total gastrectomy in the Affiliated Tumor Hospital of Zhengzhou University between May 2017 and April 2018 were collected. There were 79 males and 39 females, aged from 26 to 81 years, with an average age of 59 years. Of 118 patients, 56 patients undergoing preoperative endoscopic mucosal injection of carbon nanoparticle tracer were divided into observation group and 62 patients undergoing intraoperative serosa injection of carbon nanoparticle tracer were divided into control group. Observation indicators: the total number of lymph node dissected, the number of positive lymph node dissected, the number of lymph node dissected at the first station and the number of lymph node dissected at the second station. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed using the independent-sample t test. Measurement data with skewed distribution were represented as M (range), and comparison between groups was analyzed using the Mann-Whitney U test. Count data were described as absolute numbers, and comparison between groups was analyzed using the chi-square test or Fisher exact probability. Comparison of ordinal data was analyzed using the rank sum test. Results:The total number of lymph node dissected, the number of positive lymph node dissected, the number of lymph node dissected at the first station, the number of lymph node dissected at the second station of the observation group were 48±16, 3(range, 0-25), 26±9, 23±7, respectively. The above indicators of the control group were 41±13, 4(range, 0-28), 25±8, 16±5, respectively. There were significant differences in the total number of lymph node dissected and the number of lymph node dissected at the second station between the two groups ( t=2.494, 6.588, P<0.05), and there was no significant difference in the number of positive lymph node dissected and the number of lymph node dissected at the first station between the two groups ( Z=0.747, t=1.689, P>0.05). Conclusions:Carbon nanoparticle labeled lymph node staining using preoperative endoscopic mucosal injection of carbon nanoparticle tracer or intraoperative serosa injection of carbon nanoparticle tracer is safe and effective in total gastrectomy for gastric cancer. Compared with intraoperative serosa injection of carbon nanoparticle tracer, preoperative endoscopic mucosal injection of carbon nanoparticle tracer can increase the total number of lymph node dissected, especially the number of lymph node dissected at the second station of gastric cancer.