Objective:To clone the encoding sequence of human EBF3 gene,construct recombinant eukaryotic expression plasmid vector pEGFP/EBF3,and study the effect of EBF3 on HepG2 cell cycling.Methods:Total RNA was isolated from placental tissue.Full-length human EBF3 cDNA was amplified by RT-PCR,cloned into eukaryotic expression plasmid vector pEGFP-N1 and sequenced.The expression and sub-cellular localization of the fusion protein EBF3-EGFP in HepG2 cells were analyzed by Western blot.Cell cycles were analyzed with flow cytometry analysis.Results:Obtained full encoding sequence of early B cell factor 3 was identical with that included in GeneBank,and the eukaryotic expression plasmid vector pEGFP/EBF3 was constructed correctly.24 h after transfected by pEGFP/EBF3,the fusion protein EBF3-EGFP was observed mainly in the cellular nucleus under the inverted fluorescence microscope.Western blot analysis confirmed that the EBF3-EGFP fusion proteins of Mr 87 000 were detected in both cytoplasmic and nuclear protein of the HepG2 transfected by pEGFP/EBF3 for 24 h or 48 h.Flow cytometry analysis revealed that the percentage of cells in the S phase was markedly increased in HepG2 cells transfected by pEGFP/EBF3 as compared with that in pEGFP-N1 transfected cells.These findings suggested that transfection of EBF3 gene into HepG2 induced cell proliferation by increasing the number of cells from G1 phase to G2 phase.Conclusion:The recombinant eukaryotic expression plasmid vector pEGFP/EBF3 is successfully established.The percentage of cells in the S phase is markedly increased in pEGFP/EBF3 transfected cells as opposed to pEGFP-N1 transfected cells.It is likely that EBF3 promotes HepG2 cells proliferation through DNA replication.

A total of 192 poststroke patients with depression and anxiety were assigned randomly into study group (n =96) and control group (n =96).The study group received both capsules Shugan-jieyu and paroxetine while the control group paroxetine alone.Compared with the control group,scores of both Hamilton depression scale (HAMD) and Hamilton anxiety scale (HAMA) were significantly different at the end of week 2 and 6 in the study group (P ＜ 0.05).The HAMD and HAMA deduction rate and the scores of mangled extremity severity,mini-mental state examination and modified Barthel index at the end of week 6 of the study group were better than those of the control group (all P ＜ 0.05).The combined use of capsules Shugan-jieyu and paroxetine could improve symptoms of depression and anxiety,offer a higher safety and accelerate the rehabilitation of extremity function.

Objective To explore the influence of the combination therapy of probucol with atorvastatin on levels of serum high sensitivity C-reactive protein (hs-CRP), oxidized low-density lipoproteins (ox-LDL), and marix metallopro? teinase-9 (MMP-9) and resolution of carotid plaque in patients with acute cerebral infarction (ACI). Methods One hun? dred-six patients with acute cerebral infarction who had carotid artery color Doppler ultrasound-confirmed atherosclerot? ic plaques , were included in the present study. The patients were randomly divided into two groups: conventional treat? ment group ( 40 cases) which received atorvastatin (20mg/d) and co-treatment group (40 cases) which received Atorvas? tatin (20mg/d) and Probucol (500mg/d). Levels of hs-CRP, ox-LDL and MMP-9 were detected in all patients before treat? ment and 1, 6 and 12 months after drug therapy. The Intima-media thickness, area and numbers of carotid plaques were evaluated by using Doppler ultrasonography during a 12 months follow-up period. Results ① Serum hs-CRP and MMP-9 levels were significantly decreased at 1, 6 and 12 months after treatment, (conventional treatment group:t =14.662, 23.586, 28.179 and co-treatment group:t =47.023, 50.239, 50.774,P <0.01). The ox-LDL levels was obviously de? creased in the combined treatment group (t =4.592, 5.011, 5.892,P <0.01) but not in conventional treatment group (P > 0.05) at 1, 6 and 12 months after treatment. Serum hs-CRP, ox-LDL and MMP-9 levels were significantly lower in com? bined treatment group than in the conventional treatment group at all time points after treatment (t =7.655, 5.271, 2.492, t =4.927, 3.772, 4.673 andt =16.862, 4.251, 2.045.P <0.01 orP <0.05). ②There were not statistically differences in the IMT, plaque area and plaque numbers between these two groups before treatment (P >0.05). The IMT, plaque area and plaque numbers were significantly smaller in combined treatment group than in conventional treatment group (t =6.117, 3.290, 2.158,P <0.05). Conclusions The combination therapy of probucol with atorvastatin can greatly reduce levels of serunl hs-CRP,ox-LDL and MMP-9, indicating that the combination therapy has a strong anti-oxidant function, thereby reversing and stabilizing the atherosclerosis plaque.

Purpose: This study aimed to investigate the changes in microRNA-130a (miR-130a) and its correlation with cardiotoxicity during epirubicin/cyclophosphamide followed by docetaxel plus trastuzumab (EC-D+T) adjuvant chemotherapy in human epidermal growth factor receptor-2-positive (HER2+) breast cancer patients. Methods: A total of 72 HER2+ breast cancer patients who underwent resection and were scheduled to receive EC-D+T adjuvant therapy were consecutively enrolled. The expression of miR-130a and cardiotoxicity (defined as any of the following situations: 1) absolute decline of left ventricular ejection fraction (LVEF) ≥ 10% and LVEF < 53%; 2) heart failure; 3) acute coronary artery syndromes; and 4) fatal arrhythmia) were assessed every 3 months throughout the 15-month EC-D+T treatment. Results: The accumulating cardiotoxicity rate was 12 (16.7%), of which the incidence of heart failure, acute coronary syndrome, life-threatening arrhythmias, ΔLVEF ≥ 10%, and LVEF < 53% was 0 (0.0%), 1 (1.4%), 0 (0.0%), and 12 (16.7%), respectively. Baseline miR-130a expression was negatively correlated with LVEF (%) and positively correlated with cardiac troponin I. The expression of miR-130a gradually increased in both cardiotoxicity and noncardiotoxicity patients during EC-D+T treatment, while the increment of miR-130a was more obvious in cardiotoxicity patients compared with non-cardiotoxicity patients. Further logistic regression and receiver operating characteristic curve analysis indicated that miR-130a was an independent predictive factor for increased cardiotoxicity risk. Conclusion MiR-130a increases constantly and predicts high cardiotoxicity risk during ECD+T adjuvant chemotherapy in HER2+ breast cancer patients.

Purpose: This study aimed to investigate the changes in microRNA-130a (miR-130a) and its correlation with cardiotoxicity during epirubicin/cyclophosphamide followed by docetaxel plus trastuzumab (EC-D+T) adjuvant chemotherapy in human epidermal growth factor receptor-2-positive (HER2+) breast cancer patients. Methods: A total of 72 HER2+ breast cancer patients who underwent resection and were scheduled to receive EC-D+T adjuvant therapy were consecutively enrolled. The expression of miR-130a and cardiotoxicity (defined as any of the following situations: 1) absolute decline of left ventricular ejection fraction (LVEF) ≥ 10% and LVEF < 53%; 2) heart failure; 3) acute coronary artery syndromes; and 4) fatal arrhythmia) were assessed every 3 months throughout the 15-month EC-D+T treatment. Results: The accumulating cardiotoxicity rate was 12 (16.7%), of which the incidence of heart failure, acute coronary syndrome, life-threatening arrhythmias, ΔLVEF ≥ 10%, and LVEF < 53% was 0 (0.0%), 1 (1.4%), 0 (0.0%), and 12 (16.7%), respectively. Baseline miR-130a expression was negatively correlated with LVEF (%) and positively correlated with cardiac troponin I. The expression of miR-130a gradually increased in both cardiotoxicity and noncardiotoxicity patients during EC-D+T treatment, while the increment of miR-130a was more obvious in cardiotoxicity patients compared with non-cardiotoxicity patients. Further logistic regression and receiver operating characteristic curve analysis indicated that miR-130a was an independent predictive factor for increased cardiotoxicity risk. Conclusion MiR-130a increases constantly and predicts high cardiotoxicity risk during ECD+T adjuvant chemotherapy in HER2+ breast cancer patients.

Objective To explore the protective effects of Caspase inhibitor on in vitro hippocampal neurons of rats with oxygen glucose deprivation (OGD) injury.Methods The primary hippocampal neurons from newborn Sprague-Dawley rats were cultured for 7 days,and then,divided into control group,OGD/re-OG treatment group,Caspase inhibitor (Z-VAD-FMK) treatment group and DMSO treatment group; and OGD/re-OG was performed in the later three groups for OGD duration of 6 h.Through the experimental process,20 μmol/L Z-VAD-FMK was given to the Z-VAD-FMK treatment group,while 1‰ DMSO was added into culture medium in the DMSO group.The morphology of the neurons was observed 24 hours after re-OG under light microscopy and electron microscopy.MTT assay was used to determine the rate of survived cells and lactate dehydrogenase (LDH) content in culture medium.The neurons apoptosis rate was measured by using flow cytometry (Annexin V-FITC/PI).Caspase-3 activity of neurons was analyzed by colorimetry.The protein expression of Caspase-3 wasassessed by Western blotting.Results Cells in the OGD/re-OG treatment group showed swelled body,some having ruptured membrane,cell integrity being damaged,cell nucleus edema,mitochondrial swelling and cristae ordered chaos; cells in the Z-VAD-FMK treatment group showed decreased injury as compared with the OGD/re-OG treatment group,having cell integrity; as compared with the OGD/re-OG treatment group,Z-VAD-FMK treatment group had significantly higher absorbance value (0.204±0.019 vs.0.303±0.018) and cell survivalrate (0.481％±0.045％vs.0.704％±0.040％),obviously decreased lactate dehydrogenase (LDH) content ([406.500±21.286] U/L vs.[326.000±20.278] U/L),apoptosis rate (49.700％±3.100％ vs.29.820％±2.839％),absorbance value of the Caspase-3 active area and Caspase-3 protein expression (1.070±0.077 vs.0.785±0.058),with statistically differences (P＜0.05).Conclusions Caspase inhibitor may protect rat hippocampal neurons from oxygen glucose deprivation injury by inhibiting Caspase-3 activity.

Objective To explore the clinical efficacy of dulaglutide combined with metformin in the treatment of obese patients with type 2 diabetes mellitus(T2DM).Methods A total of 200 obese patients with T2DM who were treated in Shanghai Jiading District Anting Hospital from January 2021 to January 2023 were randomly divided into liraglutide group(n=100)and dulaglutide group(n=100).The liraglutide group was treated with liraglutide combined with metformin,and the dulaglutide group was treated with dulaglutide combined with metformin.Both groups were treated for 3 months.The body metabolic indexes[fasting blood glucose(FBG),2 h postprandial blood glucose(2 h PBG),hemoglobin(HbA1 c),total cholesterol(TC),triglyceride(TG)],body fat composition[body fat rate,body mass index,subcutaneous fat rate of limbs,visceral fat index]and serum adipokines(adiponectin,neuropeptide Q(NPQ),asprosin,irisin)levels were compared before treatment and 3 months after treatment.The clinical efficacy and adverse reactions of the two groups were observed.Results After 3 months of treatment,FBG,2 h PBG,HbAlc,TC,TG,body fat rate,body mass index,subcutaneous fat rate of limbs,visceral fat index and asprosin in the two groups were lower than those before treatment,and those in the dulaglutide group were lower than those in the liraglutide group(P＜0.05).After 3 months of treatment,the levels of serum adiponectin,NPQ and irisin in the two groups were higher than those before treatment,and the increase in the dulaglutide group was greater than that in the liraglutide group(P＜0.05).The effective rate of dulaglutide group(98.00%)was higher than that of liraglutide group(91.00%)(P＜0.05).There was no significant difference in the incidence of adverse reactions between the two groups(11.00%,14.00%)(P＞0.05).Conclusion Dulaglutide combined with metformin could improve the metabolic status of obese T2 DM patients,regulate body fat composition and serum adipokines,with significant clinical efficacy and safety.

Objective:To investigate the short-term outcomes of laparoscopic simultaneous resection of primary colorectal cancer and liver metastases in patients with resectable synchronous colorectal liver metastases (SCRLM).Methods:A descriptive case series study was performed. Clinicopathological data of patients with SCRLM who underwent laparoscopic simultaneous resection of colorectal cancer and liver metastases in Zhongshan Hospital between December 2015 and September 2018 were retrieved from a prospective colorectal cancer database. Perioperative presentations and short-term outcomes were analyzed.Results:A total of 53 patients were enrolled with average age of(61.7±11.3) years. Among them, 32 were male (60.4%) and 21 were female (39.6%). Twenty-five patients (47.2%) were American Society of Anesthesiologists (ASA) grade I and 28 (52.8%) were grade II. All the patients completed laparoscopic simultaneous resection without conversion. The average operation time was (320.2±114.5) min. The estimated blood loss was 150.0 (45.0-2000.0) ml, and only 2 cases (3.8%) received intraoperative transfusion. Postoperative pathologic results revealed that the average primary tumor size was (5.4±1.9) cm; 4 cases (7.5%) were T1-2 stage and 48 cases (90.6%) were T3-4 stage; 40 patients (75.5%) had lymph node metastasis; 19 (35.8%) had vascular involvement; 24 (45.3%) had neural invasion. The median number of liver metastases was 1.0 (1-8), and the average size of largest liver metastases was (3.0±1.9) cm. The median margin of liver metastases was 1.0 (0.1-3.5) cm, and only 1 case was R1 resection. The average time to the first postoperative flatus was (67.9±28.9) h, and the average time to the liquid diet was (107.0±33.8) h. The average postoperative indwelling catheterization time was (85.6±56.4) h. The average postoperative hospital stay was (9.2±4.4) d, and the average cost was (82±26) thousand RMB. No death within postoperative 30-day was found. The morbidity of postoperative complication was 32.1% (17/53) and 3 patients developed grade III to IV complications which were improved by conservative treatment. The median follow-up period was 23.2 months. During follow-up, 19 patients (35.8%) developed recurrence or metastasis, and 4 (7.5%) died. The 1- and 2-year disease-free survival (DFS) rates were 68% and 47% respectively, and the 1- and 2-year overall survival rates were 95% and 86% respectively.Conclusions:Laparoscopic simultaneous resection of primary colorectal cancer and liver metastases is safe and feasible in selected patients with SCRLM. Postoperative intestinal function recovery is enhanced, and morbidity and oncological outcomes are acceptable.

Objective:To investigate the short-term outcomes of laparoscopic simultaneous resection of primary colorectal cancer and liver metastases in patients with resectable synchronous colorectal liver metastases (SCRLM).Methods:A descriptive case series study was performed. Clinicopathological data of patients with SCRLM who underwent laparoscopic simultaneous resection of colorectal cancer and liver metastases in Zhongshan Hospital between December 2015 and September 2018 were retrieved from a prospective colorectal cancer database. Perioperative presentations and short-term outcomes were analyzed.Results:A total of 53 patients were enrolled with average age of(61.7±11.3) years. Among them, 32 were male (60.4%) and 21 were female (39.6%). Twenty-five patients (47.2%) were American Society of Anesthesiologists (ASA) grade I and 28 (52.8%) were grade II. All the patients completed laparoscopic simultaneous resection without conversion. The average operation time was (320.2±114.5) min. The estimated blood loss was 150.0 (45.0-2000.0) ml, and only 2 cases (3.8%) received intraoperative transfusion. Postoperative pathologic results revealed that the average primary tumor size was (5.4±1.9) cm; 4 cases (7.5%) were T1-2 stage and 48 cases (90.6%) were T3-4 stage; 40 patients (75.5%) had lymph node metastasis; 19 (35.8%) had vascular involvement; 24 (45.3%) had neural invasion. The median number of liver metastases was 1.0 (1-8), and the average size of largest liver metastases was (3.0±1.9) cm. The median margin of liver metastases was 1.0 (0.1-3.5) cm, and only 1 case was R1 resection. The average time to the first postoperative flatus was (67.9±28.9) h, and the average time to the liquid diet was (107.0±33.8) h. The average postoperative indwelling catheterization time was (85.6±56.4) h. The average postoperative hospital stay was (9.2±4.4) d, and the average cost was (82±26) thousand RMB. No death within postoperative 30-day was found. The morbidity of postoperative complication was 32.1% (17/53) and 3 patients developed grade III to IV complications which were improved by conservative treatment. The median follow-up period was 23.2 months. During follow-up, 19 patients (35.8%) developed recurrence or metastasis, and 4 (7.5%) died. The 1- and 2-year disease-free survival (DFS) rates were 68% and 47% respectively, and the 1- and 2-year overall survival rates were 95% and 86% respectively.Conclusions:Laparoscopic simultaneous resection of primary colorectal cancer and liver metastases is safe and feasible in selected patients with SCRLM. Postoperative intestinal function recovery is enhanced, and morbidity and oncological outcomes are acceptable.

ObjectiveTo explore the possible mechanism of Pingwei Capsules （平胃胶囊） for chronic atrophic gastritis from rapidly accelerated fibrosarcoma / mitogen-activated protein kinase /extracellular-signal-regulated kinase （Raf/MEK/ERK） pathway that influences the activation of fibrosarcoma protein/mitogen. MethodsFifteen SD rats were randomly divided into 5 rats in the blank group and 10 rats in Pingwei Capsules group. The rats in the blank group were given 1 ml/100 g of saline by gavage， and the rats in Pingwei Capsules group were given 0.63 g/（kg·d） of Pingwei Capsule suspension by gavage， and serum was collected for 3 consecutive days. N-methyl-N'-nitro-N-nitrosoguanidine （MNNG） was used to induce human gastric mucosal epithelial cells GES-1 to establish a precancerous lesion cell model. The successful cells were divided into control group （10% fetal bovine serum）， blank serum group （10% fetal bovine serum plus 10% blank serum）， and medication-containing serum group （serum with medication of Pingwei Capsule）， and the volume fraction and time of intervention of Pingwei Capsule-containing serum were screened by CCK-8 assay. Human gastric mucosal epithelial cells GES-1 were divided into normal group， model group， blank serum group， medication-containing serum group， U0126 group， and combined group， with 6 replicate wells in each group. After successful modelling of the cells in all groups except the blank group， an equal volume of fetal bovine serum was added to the normal and model groups， an equal volume of blank serum was added to the blank serum group， a screening volume fraction of Pingwei Capsule-containing serum was added to Pingwei Capsule group， a 10 μmol/L mitogen-activated extracellular signal regulated kinase 1 （MEK1） inhibitor U0126 was administered in the U0126 group， an equal dose of Pingwei Capsule-containing serum plus 10 μmol/L of U0126 was administered to the combined group. After the selected incubation time， the level of interleukin 6 （IL-6） was detected in the cells by ELISA， the expression of IL-6 and MEK1 was detected by immunofluorescence， and the expression of IL-6， Raf， MEK1， and ERK mRNA was detected by RT-qPCR， and the expression of IL-6， Raf， MEK1， and ERK mRNA in the cells was detected by Western blot. ResultsThe 5.35% volume fraction， 48 h intervention of Pingwei Capsule-containing serum was selected for subsequent experiments. Compared with the normal group， the IL-6 content in cell supernatants and the expression of IL-6， Raf， MEK1， ERK mRNA and ERK1/2 proteins in cells increased in the model group and blank serum group （P＜0.01）. Compared with the model group， all of the above indexes were improved in medication-containing serum group， U0126 group， and combined group （P＜0.05 or P＜0.01）. Compared with medication-containing serum group， the expression of IL-6， MEK1 expression， the expression of IL-6， Raf， MEK1 and ERK mRNA， and the expression of IL-6， Raf， MEK1 and ERK1/2 proteins reduced in the cells of combined group （P＜0.05 or P＜0.01）. Compared with the U0126 group， IL-6 expression reduced and IL-6， MEK1 and ERK1/2 protein expression reduced in cells of combined group （P＜0.05 or P＜0.01）. ConclusionThe Pingwei Capsule-containing serum may play a role in the treatment of chronic atrophic gastritis by improving the inflammation-cancer transformation of GES-1 cells through inhibiting the Raf/MEK/ERK pathway.