Alfalfa (Medicago sativa) is an important food and feed crop which rich in mineral sources. The WUSCHEL-related homeobox (WOX) gene family plays important roles in plant development and identification of putative gene families, their structure, and potential functions is a primary step for not only understanding the genetic mechanisms behind various biological process but also for genetic improvement. A variety of computational tools, including MAFFT, HMMER, hidden Markov models, Pfam, SMART, MEGA, ProtTest, BLASTn, and BRAD, among others, were used. We identified 34 MsWOX genes based on a systematic analysis of the alfalfa plant genome spread in eight chromosomes. This is an expansion of the gene family which we attribute to observed chromosomal duplications. Sequence alignment analysis revealed 61 conserved proteins containing a homeodomain. Phylogenetic study sung reveal five evolutionary clades with 15 motif distributions. Gene structure analysis reveals various exon, intron, and untranslated structures which are consistent in genes from similar clades. Functional analysis prediction of promoter regions reveals various transcription binding sites containing key growth, development, and stress-responsive transcription factor families such as MYB, ERF, AP2, and NAC which are spread across the genes. Most of the genes are predicted to be in the nucleus. Also, there are duplication events in some genes which explain the expansion of the family. The present research provides a clue on the potential roles of MsWOX family genes that will be useful for further understanding their functional roles in alfalfa plants.

Objective:To construct a home-based cardiac rehabilitation intervention system for patients after percutaneous coronary intervention, and to provide reference for improving the self-management ability and family support of home-based cardiac rehabilitation of patients after PCI.Methods:Based on the literature study and group discussions, a draft of home-based cardiac rehabilitation intervention system for patients after PCI based on empowerment theory was constructed. From January to April 2021, the Delphi method was used to conduct 2 rounds of expert consultations among 18 experts from 9 hospitals, and the items were modified according to the experts′ advice.Results:The expert positive coefficients of the 2 rounds were 94.44% and 100.00%, the expert authority coefficients was 0.91, and the Kendall coefficients were 0.188 and 0.255. Finally, a home-based cardiac rehabilitation intervention system for patients after PCI was formed, including 5 first-level items, 19 second-level items and 21 third-level items.Conclusion:The home-based cardiac rehabilitation intervention system for patients after PCI is reliable, scientificity and practical, and has guiding significance for promoting the development of home-based cardiac rehabilitation for PCI patients.

Objective To construct the prokaryotic expression vector of human esophageal cancer related gene 4 (ECRG4), to purify the recombinant ECRG4 protein and to verify the biological function of the recombinant ECRG4 protein. Methods DNA recombination technology was utilized to construct the ECRG4 protein prokaryotic expression vector. The recombinant ECRG4 protein was purified with the transformation of escherichia coli. Then the purity of the recombinant ECRG4 protein was examined by using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. Furthermore, esophageal cancer EC-18 cell line was treated by recombinant ECRG4 protein (10 μg/ml) for phosphate buffer (PBS) 48 h respectively, and the tumor cell cycle change was examined by using flow cytometry. Results SDS-PAGE analysis showed that the high purity of recombinant ECRG4 protein was obtained and the ECRG4 protein prokaryotic expression vector was successfully constructed. The cell proportion of G1 phase in PBS control group was lower than that in the recombinant ECRG4 protein group [(60.4 ±2.1) ％ vs. (71.6 ±1.8) ％; t= 25.695, P= 0.002]. The cell proportion of S phase in PBS control group was higher than that in the recombinant ECRG4 protein group [(24.6±1.4) ％ vs. (16.5±1.0) ％; t= 36.905, P= 0.001]. The cell proportion of G2/M phase in PBS control group was higher than that in the recombinant ECRG4 protein group [(15.0 ±1.1) ％ vs. (11.9 ±0.8) ％; t=6.471, P=0.023], which indicated that the recombinant ECRG4 protein could induce the G1 phase arrest of EC-18 cells. Conclusion The ECRG4 protein prokaryotic expression vector is successfully constructed. And the recombinant ECRG4 protein has an active biological function in esophageal carcinoma.

Objective:To investigate the risk factors that were associated with the death of elderly patients who were admitted to the intensive care unit (ICU) after elective abdominal surgery, and to find reliable and sensitive predictive indicators for early interventions and reducing the mortality.Methods:A retrospective case-control study was conducted. The clinical data of elderly (age≥65 years old) patients after elective abdominal surgery admitted to the ICU of the Affiliated Hospital of Guizhou Medical University from January 1st 2016 to December 31st 2020 were collected, including the patient's gender, age, body mass index (BMI), medical history, American Society of Anesthesiologists (ASA) grades, surgical classification, intraoperative blood loss, duration of operation, interval time between end of operation and admission to the ICU, acute physiology and chronic health evaluationⅡ(APACHEⅡ) score and the worst laboratory examination results within 24 hours of ICU admission, the first blood gas analysis in ICU, the duration of invasive mechanical ventilation, and the length of ICU stay. Postoperative abdominal infection was evaluated by the pathogenic culture of peritoneal drainage fluid and clinical symptoms and signs. The patients were divided into death group and survival group based on clinical outcomes, and clinical data were compared between the two groups. Binary multivariate Logistic regression analysis was used to screen the risk factors of death, and the receiver operator characteristic curve (ROC curve) was plotted to analyze the predictive values of these risk factors.Results:A total of 226 elderly patients with elective abdominal surgery were admitted to the ICU of our hospital during the past 5 years, of whom, two patients who did not undergo laboratory examinations within 24 hours of admission to the ICU were excluded. Finally, 224 patients met the criteria, with 158 survivors and 66 deaths. Univariate analysis showed that: compared with survival group, APACHEⅡscore, blood lactate acid (Lac) and the proportion of postoperative abdominal infection were higher in death group [APACHEⅡ score: 27.5 (25.0, 31.3) vs. 23.0 (18.0, 27.0), Lac (mmol/L): 2.9 (1.8, 6.6) vs. 1.8 (1.1, 2.8), the proportion of postoperative abdominal infection: 65.2% (43/66) vs. 35.4% (56/158), all P < 0.01], prothrombin time (PT), activated partial thromboplastin time (APTT) and interval time between end of surgery and admission to ICU were longer [PT (s): 17.20 (14.50, 18.63) vs. 14.65 (13.90, 16.23), APTT (s): 45.15 (38.68, 55.15) vs. 39.45 (36.40, 45.70), interval time between end of surgery and admission to ICU (hours): 39.2 (0.7, 128.9) vs. 0.7 (0.3, 2.0), all P <0.01], postoperative hemoglobin (Hb), platelet count (PLT), prealbumin (PA), mean arterial pressure (MAP) and oxygenation index (PaO 2/FiO 2) were lower in death group [Hb (g/L): 95.79±23.64 vs. 105.58±19.82, PLT (×10 9/L): 138.5 (101.0, 177.5) vs. 160.5 (118.5, 232.3), PA (g/L): 80.88±43.63 vs. 116.54±50.80, MAP (mmHg, 1 mmHg = 0.133 kPa): 76.8±19.1 vs. 91.6±19.8, PaO 2/FiO 2 (mmHg): 180.0 (123.5, 242.5) vs. 223.5 (174.8, 310.0), all P < 0.05]. Binary multivariate Logistic regression analysis showed that APACHEⅡscore [odds ratio ( OR) = 1.187, 95% confidence interval (95% CI) =1.008-1.294, P < 0.001], interval time between end of operation and admission to ICU ( OR = 1.005, 95% CI = 1.001-1.009, P = 0.016) and postoperative abdominal infection ( OR = 2.630, 95% CI = 1.148-6.024, P = 0.022) were independent risk factors for prognosis in these patients. MAP ( OR = 0.978, 95% CI = 0.957-0.999, P = 0.041) and PaO 2/FiO 2 ( OR = 0.994, 95% CI = 0.990-0.998, P = 0.003) were protective factors for the patients' prognosis. Lac, Hb, PLT, PA, PT and APTT had no predictive value for the prognosis of elderly patients admitted to ICU after elective abdominal surgery [ OR value and 95% CI were 1.075 (0.945-1.223), 1.011 (0.99-1.032), 1.000 (0.995-1.005), 0.998 (0.989-1.007), 1.051 (0.927-1.192) and 1.003 (0.991-1.016), respectively, all P > 0.05. ROC curve analysis showed that APACHEⅡscore, interval time between end of operation and admission to the ICU and the postoperative abdominal infection had certain predictive values for the prognosis of elderly patients, the area under ROC curve (AUC) were 0.755, 0.732 and 0.649 respectively, all P < 0.001; When the cut-off of APACHEⅡscore and interval time between end of operation and admission to the ICU were 24.5 scores and 2.15 hours, the sensitivity were 78.8% and 66.7%, respectively, and the specificity were 62.0% and 76.6%, respectively. The combined predictive value of the three variables was the highest, which AUC was 0.846, the joint prediction probability was 0.27, the sensitivity was 83.3%, and the specificity was 75.3%. Conclusion:APACHEⅡscore, interval time between end of surgery and admission to ICU, and postoperative abdominal infection may be independent risk factors for the death of elderly patients who were admitted to the ICU after elective abdominal surgery, there would be far greater predictive values when the three variables were combined.

OBJECTIVETo evaluate the prognosis and predictive values of preoperative Glasgow prognostic score (GPS) for adenocarcinoma of esophagogastric junction(AEG) patients.

METHODSA retrospective study of 322 AEG patients who received operation between January 2007 and March 2010 in Henan Provincial People's Hospital was performed. Clinical data, pathological characteristics, laboratory parameters and survival data were collected. The GPS was calculated based on C-reactive protein(CRP) and serum albumin(ALB) levels. Univariate and multivariate analysis were used to evaluate the prognostic value of GPS.

RESULTSAmong 322 patients, 0, 1, 2 of GPS were 192, 104 and 26 patients respectively. The median follow-up was 37 (4-73) months. In Kaplan-Meier analysis, median diseases-free survival (DFS) of GPS 0, 1, 2 was 47.0 (95% CI: 31.6-62.4), 15.0 (95% CI: 11.8-8.2) and 4.7 (95% CI: 3.8-5.6) months (P<0.01), and median overall survival (OS) was out of reach, 20.6 (95% CI: 15.8-25.4) and 7.0 (95% CI: 5.8-8.2) months (P<0.01). Univariate and multivariate analysis revealed that GPS was an independent predictor of DFS (P<0.01) and OS (P<0.01) of AEG.

CONCLUSIONGPS is an effective predictor of survival in AEG.

Adenocarcinoma ; C-Reactive Protein ; Disease-Free Survival ; Esophageal Neoplasms ; Esophagogastric Junction ; Humans ; Kaplan-Meier Estimate ; Prognosis ; Retrospective Studies ; Stomach Neoplasms

Objective: To study the predictive and prognostic significance of high-sensitivity modified Glasgow Prognostic Score (HS-mGPS) on the effect of neoadjuvant chemotherapy for advanced gastric cancer. Methods: 117 patients with advanced gastric cancer received neoadjuvant chemotherapy with SOX (oxaliplatin+ S1) or mFOLFOX 6(oxaliplatin+ CF+ 5-FU) regimen. HS-mGPS was calculated according to blood C-reactive protein (CRP) concentration and serum albumin (ALB) level. The correlation between HS-mGPS and clinicopathological characteristics was determined and the predictors of survival were analyzed. Results: 117 patients with stage ⅡB (43 cases), stage Ⅲ (60), and stage Ⅳ (14) received preoperative neoadjuvant chemotherapy. The overall response rate of neoadjuvant chemotherapy was 61.5%(72/117), and the tumor control rate was 88.0% (103/117), with a pathological response rate of 91.5% (107/117). The R0 resection rate was 81.2% (95/117). The median disease-free survival (DFS) was 21.0 (95% CI 6.4-35.6) months. The median overall survival (OS) was 39.0 (95% CI 21.4-56.6) months. Higher HS-mGPS was associated with higher T stage, local lymph-node metastasis, distant metastasis, lower chemotherapy overall response rate and lower pathological response rate (all P<0.05). The univariate analysis and multivariate analysis showed that higher HS-mGPS, presence of local lymph-node metastasis and non R0 resection were associated with poorer DFS and OS (P<0.05). Conclusion HS-mGPS can be used to predict the benefits of neoadjuvant chemotherapy and as an independent prognostic factor for survival in patients with advanced gastric cancer.

Objective : To investigate the effect of Arginine Vasopressin (AVP) on the median preoptic glutamatergic (MnPOVglut2 ) neurons and its mechanism． Methods : Brain slices were prepared from male Vglut2-tdTomato mice．MnPOVglut2 neurons expressing red fluorescent protein were located by using fluorescence microscope．Wholecell patch clamp technique was used to observe the effect of AVP on the firing frequency of MnPOVglut2 neurons，the effect of synaptic transmission blockers ( STBs) on the AVP-induced change in the firing frequency of MnPOVglut2 neurons，and the effect of AVP V1a receptor antagonist on the AVP-induced change in the firing frequency of MnPOVglut2 neurons. Results: The mean firing frequency of MnPOVglut2 neurons increased during perfusion with artificial cerebrospinal fluid (ACSF) and AVP compared with that during perfusion with ACSF (P＜0. 01) ，indicating that AVP excited the MnPOVglut2 neurons．The mean firing frequency of MnPOVglut2 neurons still increased during perfusion with ACSF，STBs，and AVP compared with that during perfusion with ACSF and STBs (P＜0. 001) ; moreover，the magnitude of AVP-induced increase in firing frequency didn't change significantly during perfusion with ACSF，STBs，and AVP compared with that during perfusion with ACSF and AVP (P ＞0. 05 ) ，suggesting that AVP excited the MnPOVglut2 neurons directly in a postsynaptic manner．The magnitude of AVP-induced increase in the firing frequency of MnPOVglut2 neurons declined during perfusion with ACSF，STBs，AVP，and V1areceptor antagonist compared with that during perfusion with ACSF，STBs，and AVP (P＜0. 01) ，suggesting that AVP excited MnPOVglut2 neurons directly via V1a receptor． Conclusion AVP can excite MnPOVglut2 neurons via V1areceptor directly in a postsynaptic manner．This study reveals the molecular marker of MnPO neurons which AVP act on.

BACKGROUND: Colorectal carcinogenesis and progression are related to the gut microbiota and the tumor immune microenvironment. Our previous clinical trial demonstrated that berberine (BBR) hydrochloride might reduce the recurrence and canceration of colorectal adenoma (CRA). The present study aimed to further explore the mechanism of BBR in preventing colorectal cancer (CRC). METHODS: We performed metagenomics sequencing on fecal specimens obtained from the BBR intervention trial, and the differential bacteria before and after medication were validated using quantitative polymerase chain reaction. We further performed ApcMin/+ animal intervention tests, RNA sequencing, flow cytometry, immunohistochemistry, and enzyme-linked immunosorbent assays. RESULTS: The abundance of fecal Veillonella parvula ( V . parvula ) decreased significantly after BBR administration ( P = 0.0016) and increased through the development from CRA to CRC. Patients with CRC with a higher V. parvula abundance had worse tumor staging and a higher lymph node metastasis rate. The intestinal immune pathway of Immunoglobulin A production was activated, and the expression of TNFSF13B (Tumor necrosis factor superfamily 13b, encoding B lymphocyte stimulator [BLyS]), the representative gene of this pathway, and the genes encoding its receptors (interleukin-10 and transforming growth factor beta) were significantly upregulated. Animal experiments revealed that V. parvula promoted colorectal carcinogenesis and increased BLyS levels, while BBR reversed this effect. CONCLUSION: BBR might inhibit V. parvula and further weaken the immunomodulatory effect of B cells induced by V. parvula , thereby blocking the development of colorectal tumors. TRIAL REGISTRAION ClinicalTrials.gov, No. NCT02226185.
Animals ; Humans ; Berberine/therapeutic use* ; Carcinogenesis ; Veillonella ; Colorectal Neoplasms/genetics* ; Tumor Microenvironment