Objective To investigate the molecular mechanism of Brusatol(BRU)regulating apoptosis of Hela cells through TLR9-MyD88 signaling pathway.Methods Hela cells preserved in our laboratory were treated with Brucea javanica at different concentrations(0,5.0,10.0,20.0,40.0,80.0 nmol·L-1).Hela cells were divided into Control group(normal cultured Hela cells),BRU-L group(treated with 10.0 nmol·L-1 Brucea javanica)and BRU group-H(treated with 20.0 nmol·L-1 Brucea javanica)Cells were treated with nmol·L-1 of Brucea javanica),BRU+pcDNA-NC group(transfected with pcDNA-NC+20.0 nmol·L-1 of Brucea javanica),BRU-H+pcDNA-TLR9 group(transfected with pcDNA-TLR9+20.0 nmol·L-1 of Brucea javanica).Cell proliferation was detected by CCK-8 and EdU methods.Apoptosis was detected by TUNNEL staining and flow cytometry.The protein expression levels of TLR9,MyD88,Bax,Bcl-2,Cleaved Caspase-3 and Cleaved Caspase-9 were detected by Western blot.Cell apoptosis and mitochondrial membrane potential detection kit(JC-1)were detected by flow cytometry,and the contents of adenosine triphosphate(ATP)and reactive oxygen species(ROS)were detected by ELISA.Results Compared with 0 nmol·L-1 group,the survival rate and IC50 value of 10 nmol·L-1 group were significantly decreased(P<0.05).After stimulation of BRU with different concentrations,the proliferation ability of cells was significantly decreased in a dose-dependent manner(P<0.05).Compared with control group,the 5-ethynyl-2'-deoxyuracil(EDU)positive cell rate,TUNEL positive cell rate,apoptosis rate and Bcl-2 protein of cells in BRU-L,BRU-H and pcDNA-NC groups were significantly decreased.The protein levels of TLR9 and MyD88,Bax,Bax/Bcl-2,Cleaved Caspase-3 and Cleaved Caspase-9 were significantly increased(P<0.05).In control group,BRU-L,BRU-H group/BRU-H+pcDNA-NC,there was a continuous decreasing trend(P<0.05).Compared with the BRU-H+pcDNA-NC group,the EDU positive cell rate,TUNEL positive cell rate,apoptosis rate and Bcl-2 protein in the BRU-H+pcDNA-TLR9 group were significantly increased.The protein levels of TLR9 and MyD88,Bax,Bax/Bcl-2,Cleaved Caspase-3 and Cleaved Caspase-9 were significantly decreased(P<0.05).Compared with Control group,JC-1 level and ATP content in BRU-L,BRU-H and BRU-H+pcDNA-NC groups were significantly decreased,while ROC content and mitotracker positive cell level were significantly increased(P<0.05).Compared with BRU-L group,JC-1 level and ATP content in BRU-H group and BRU-H+pcDNA-NC group were further decreased,while ROC content and mitotracker positive cell level were further increased(P<0.05).Compared with the BRU-H+pcDNA-NC group,the levels of JC-1 and ATP in the BRU-H+pcDNA-TLR9 group were increased,while the levels of ROC,mitotracker staining positive cells were decreased(P<0.05).Conclusion Brucea javanica can produce Hela cell proliferation by regulating TLR9-MyD88 signaling pathway.

Anti-seizure medications (ASMs) are the main therapy for epilepsy.There are many kinds of ASMs with complex mechanism of action, so it is difficult for pharmacists to examine prescriptions.This paper put forward some suggestions on the indications, dosage forms/routes of administration, appropriateness of usage and dosage, combined medication and drug interaction, long-term prescription review, individual differences in pathophysiology of children, and drug selection when complicated with common epilepsy, for the reference of doctors and pharmacists.

Objective:To analyze the efficacy of nutrition intervention combined with rehabilitation exercise in elderly patients with chronic heart failure (CHF) complicated by sarcopenia.Methods:In this randomized controlled trial, seventy CHF patients with sarcopenia, aged 80 years or older, who had received standard CHF medication treatment and had stable conditions with preserved ejection fraction (HFpEF) from June 2021 to October 2022 at the China-Japan Friendship Hospital outpatient clinic were continuously selected. The patients were randomly divided into a test group ( n=35) and a control group ( n=35) using a random number table. The test group continued the original heart failure medication treatment, and on the basis of daily meal, supplemented with whey protein powder [37 g per serving (including 16.2 g of whey protein, 145 kcal), twice a day]; they were given resistance exercise primarily focused on resistance training (5 sets of exercises, progressively increasing to 3 days per week, twice a day, 5 sets per session, 20 repetitions per set). The control group maintained the original heart failure medication treatment plan and lifestyle. The body mass index, appendicular skeletal muscle mass index (ASMI), grip strength, Five-Times-Sit-to-Stand Test, 6-meter walking speed, 6-minute walk test, brain natriuretic peptide (BNP), left ventricular ejection fraction, ratio of early mitral flow peak velocity to early mitral annulus diastolic peak velocity (E/e′), serum albumin, hemoglobin, and serum creatinine were compared between the two groups before and after 3 months of intervention using t-tests and chi-square tests to analyze the efficacy of nutrition intervention combined with rehabilitation exercise in elderly patients with CHF complicated by sarcopenia. Results:Among the 70 patients, there were 42 males and 28 females. There was no significant difference in baseline data between the test group and the control group (all P>0.05). After 3 months of intervention, ASMI, grip strength and 6 min walking distance in the test group were all significantly higher than those in the control group [(6.43±1.07) vs (6.09±0.86) kg/m 2, (27.75±2.13) vs (23.45±0.47) kg, (361.51±58.71) vs (273.50±69.85) m], and significantly higher than those before intervention [(5.99±1.45) kg/m 2, (23.55±9.64) kg, (273.50±69.86) m]. The time of Five-Times-Sit-to-Stand Test, BNP and E/e′ in the test group were both significantly lower than those in the control group [(11.20±2.09) vs (15.36±0.46) s, (278.80±58.69) vs (400.80±87.86) ng/L, (11.10±0.81) vs (14.66±1.90)], and significantly lower than those before intervention [(18.51±2.90) s, (407.50±122.74) ng/L and (14.00±1.15)]. There was no significant difference in remaining indicators between the two groups (all P>0.05). Conclusion:Nutritional intervention combined with rehabilitation exercise primarily focused on whey protein supplementation and resistance training can significantly improve limb muscle mass, muscle strength, muscle function, and cardiac function in elderly patients with CHF complicated by sarcopenia on the basis of heart failure medication treatment.

Objective:To analyze the risk factors of bronchopulmonary dysplasia(BPD)in very preterm infants(VPI), and to provide scientific basis for the prevention and treatment of BPD in VPI.Methods:A prospective multicenter study was designed to collect the clinical data of VPI in department of neonatology of 28 hospitals in 7 regions from September 2019 to December 2020.According to the continuous oxygen dependence at 28 days after birth, VPI were divided into non BPD group and BPD group, and the risk factors of BPD in VPI were analyzed.Results:A total of 2 514 cases of VPI including 1 364 cases without BPD and 1 150 cases with BPD were enrolled.The incidence of BPD was 45.7%.The smaller the gestational age and weight, the higher the incidence of BPD( P<0.001). Compared with non BPD group, the average birth age, weight and cesarean section rate in BPD group were lower, and the incidence of male infants, small for gestational age and 5-minute apgar score≤7 were higher( P<0.01). In BPD group, the incidences of neonatal respiratory distress syndrome(NRDS), hemodynamically significant patent ductus arteriosus, retinopathy of prematurity, feeding intolerance, extrauterine growth restriction, grade Ⅲ~Ⅳ intracranial hemorrhage, anemia, early-onset and late-onset sepsis, nosocomial infection, parenteral nutrition-associated cholestasis were higher( P<0.05), the use of pulmonary surfactant(PS), postnatal hormone exposure, anemia and blood transfusion were also higher, and the time of invasive and non-invasive mechanical ventilation, oxygen use and total hospital stay were longer( P<0.001). The time of starting enteral nutrition, cumulative fasting days, days of reaching total enteral nutrition, days of continuous parenteral nutrition, days of reaching 110 kcal/(kg·d) total calorie, days of reaching 110 kcal/(kg·d) oral calorie were longer and the breastfeeding rate was lower in BPD group than those in non BPD group( P<0.001). The cumulative doses of amino acid and fat emulsion during the first week of hospitalization were higher in BPD group( P<0.001). Multivariate Logistic regression analysis showed that NRDS, invasive mechanical ventilation, age of reaching total enteral nutrition, anemia and blood transfusion were the independent risk factors for BPD in VPI, and older gestational age was the protective factor for BPD. Conclusion:Strengthening perinatal management, avoiding premature delivery and severe NRDS, shortening the time of invasive mechanical ventilation, paying attention to enteral nutrition management, reaching whole intestinal feeding as soon as possible, and strictly mastering the indications of blood transfusion are very important to reduce the incidence of BPD in VPI.

Objective:To investigate the expression and correlation of H19, matrix metalloproteinase (MMP)-2 and MMP-9 in patients with recurrent spontaneous abortion (RSA).Methods:Human extravillous trophoblast cell line HTR-8 was cultured in vitro. Lentivirus was used to infect the HTR-8 cell line to over-express or knockdown the expression of H19. The concentrations of MMP-2 and MMP-9 protein in cell culture supernatant were detected by enzyme linked immunosorbent assay (ELISA). The expression levels of H19, MMP-2 and MMP-9 mRNA in villi of patients with RSA were detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). Spearman correlation analysis was used to understand the correlation between H19 and the expression levels of MMP-2 and MMP-9. Results:After overexpression of H19, the expression levels of MMP-2 and MMP-9 mRNA and protein concentration in Lv-ph19 group were significantly higher than those in Lv-vector group ( P<0.05); After interfering with the expression of H19, the expression levels of MMP-2 and MMP-9 mRNA and protein concentration in Lv-shH19 group were significantly lower than those in Lv-shcon control group ( P<0.05). The number of spontaneous abortions in patients with recurrent spontaneous abortion was significantly higher than that in the control group ( P<0.05). qRT-PCR showed that the expression levels of H19, MMP-2 and MMP-9 mRNA in villi of patients with RSA were significantly lower than those in the control group ( P<0.05). There was a positive correlation between H19 and the expression levels of MMP-2 and MMP-9 ( P<0.05). Conclusions:H19 regulates the expression of MMP-2 and MMP-9 of trophoblast during early pregnancy, and the abnormal expression of H19, MMP-2, and MMP-9 in human first-trimester villous tissues was related with the incidence of early miscarriage.

Objective To investigate the effects of β-Elemene on the apoptosis of human pancreatic cancer Panc-1 cells; To discuss its mechanism of action. Methods β-Elemene (10, 20, 40, 80, 160 μg/mL) were incubated to the Panc-1 cells in vitro cultured for 24 h, 48 h and 72h, and trypan blue refusal method was used to detect cell inhibition rate;Apoptosis rate was measured by TUNEL; Hoechst33258 fluorescent staining was used to observe the changes of the nucleus. The activity of Caspase-3, 8 and 9 were detected by ELISA. Western blot was used to detect the expressions of Fas, FasL and Cyt c and AIF. Results The activity of Panc-1 cells was obviously inhibited time/concentration dependent inhibition (P<0.05, P<0.01), and the apoptosis rate increased after incubated with β-Elemene (P<0.01, P<0.001) after incubated with β-Elemene for 24 h, 48 h and 72 h; After giving β-Elemene 72 h, Panc-1 cells nucleus were broken obviously, and chromatin condensed and showed strong blue fluorescence, along with of apoptotic bodies; After incubated with β-Elemene for 48 h, Caspase-3, 8 and 9 activity significantly increased (P<0.05, P<0.01); protein expressions of Fas, FasL, Cyt c and AIF were significantly enhanced (P<0.05, P<0.01, P<0.001). Conclusion β-Elemene can inhibit Panc-1 cell proliferation, induce apoptosis, and the mechanism may be related to activating cell death receptor pathway and mitochondrial apoptosis pathway to play anti-tumor effects.

OBJECTIVETo evaluate and compare the value of stair climbing tests and conventional pulmonary function tests in the prediction of postoperative cardiopulmonary complications in non-small cell lung cancer patients underwent surgery.

METHODSFrom April 1, 2010 to Jan. 30, 2012, a total of 162 patients with thoracic carcinoma underwent stair climbing test (SCT) and conventional pulmonary function tests (PFT) preoperatively. The correlation of postoperative cardiopulmonary complications with the SCT and PFT parameters were analyzed retrospectively using chi-square test, independent sample t test and binary logistic regression analysis.

RESULTSOf the 162 patients, 19 without operation were excluded, due to an advanced stage (9 cases), poor cardiopulmonary function (5 cases), rejecting operation (4 cases) and exploration alone (1 case). 143 cases were eligible and evaluated eventually. Forty-one of the 143 patients (28.7%) had postoperative cardiopulmonary complications, but no death occurred. The patients were stratified into groups based on the time of stair climbing 5 stories (18.36 m, t, <92 s, ≥ 92 s). Exercise oxygen desaturation (EOD) during the stair climbing test (<5%, ≥ 5%) and the difference between the pulse at resting state and the pulse at end of stair climbing test (ΔP, <55 beats/min, ≥ 55 beats/min), respectively. The rate of postoperative cardiopulmonary complications was significantly higher in the group with t ≥ 92 s, EOD ≥ 5% and ΔP < 55 beats/min (38.5%, 42.0% and 35.1%, respectively) than that in the group with t<92 s, EOD<5% and ΔP ≥ 55 beats/min (16.9%, 21.5% and 18.2%, respectively). Binary logistic regression analysis showed that postoperative cardiopulmonary complications were independently correlated with EOD and lung function which did not meet the requirement of the lung resection operation mode.

CONCLUSIONSA symptom-limited stair climbing test is a safe, simple and low-cost method to evaluate the cardiopulmonary function preoperatively. It can predict the occurrence of postoperative cardiopulmonary complications in non-small cell lung cancer patients. Conventional pulmonary function tests and stair-climbing test can be recommended to be routinely performed in all patients with non-small cell lung cancer before thoracic surgery.

Carcinoma, Non-Small-Cell Lung ; surgery ; Exercise Test ; Humans ; Lung Neoplasms ; surgery ; Postoperative Complications ; diagnosis ; Respiratory Function Tests ; Retrospective Studies

BACKGROUND:In vitro studies have demonstrated that basic fibroblast growth factor (bFGF) promote the differentiation of bone marrow mesenchymal stem cells (BMSCs) into cardiomyocyte-like cells. However, it is unclear whether coronary venous retroperfusion of bFGF stimulates BMSCs differentiation in vivo. OBJECTIVE:To evaluate the effects of coronary venous retroperfusion of bFGF on BMSCs differentiation in vivo. METHODS:BMSCs from 12 dogs were isolated by density gradient centrifugation and expanded in vitro. These cells were transfected by enhanced green fluorescence protein (EGFP) lentiviral vector and the transfection efficiency was analyzed. Acute myocardial infarction was induced by ligation of left anterior descending coronary artery. After 1 week, 10 survival animals were randomized to BMSCs group (n=5) and bFGF+BMSCs group (n=5). bFGF-and EGFP-positive BMSCs were reversely infused via coronary vein using over-the-wire bal oon catheter. One week after infusion, the number of EGFP-positive cells co-staining factor VIII and troponin I was compared between the two groups by immunofluorescence method. RESULTS AND CONCLUSION:BMSCs were successful y transfected by EGFP and the transfection efficiency was 85%. Immunofluorescence showed that EGFP-positive BMSCs were observed in 23.5%of slides. There were more EGFP-positive cells co-staining VIII and troponin I in the bFGF+BMSCs group than in the BMSCs group (P<0.05). Thus, the coronary venous retroperfusion of bFGF enhances the differentiation of BMSCs into vascular endothelial cells and cardiomyocytes. Combined delivery of bFGF and BMSCs can exert a synergistic effect to promote cardiac repair.

10.3969/j.issn.2095-4344.2013.24.015

ObjectiveTo study the impact of primary PCI in culprit artery on epicardial blood flow of nonculprit artery in patients with STEMI. MethodsEnrolled 117 patients with anterior wall STEMI were treated with primary PCI in the culprit artery, left anterior descending artery (LAD, as study group.Another 100 patients with normal coronary artery evidenced by angiography were enrolled as control group.The differences in CTFC (corrected TIMI frame count measured by using digital subtraction arteriography,TIMI =thrombolysis in myocardial infarction) and MBG (myocardium blood flow perfusion grading)between pre and post primary PCI in both culprit artery and nonculprit artery ( left circumflex artery, LCX),and CTFC and MBG were also detected in the subjects of control group. Blood samples were collected and the levels of CRP (C-reactive protein) were assayed. Clinical and angiographic features were analyzed.ResultsThe CTFC of nonculprit artery (LCX) and the level of MBG in patients with anterior wall STEMI were different from the level of MBG and CTFC in control group ( P＜0. 05) before primary PCI. The level of MBG and CTFC in nonculprit artery (LCX) were improved (P ＜ 0. 05 ) after primary PCI, but they did not resume to normal level. Patients without reflow in culprit artery had higher incidence of no reflow in nonculprit artery than patients with re-flow (78％ vs. 8％, P ＜ 0. 01 ), and the level of CRP in patients without reflow in nonculprit artery were higher than those in patients with re-flow ( P ＜ 0. 05). Conclusions The perfusion of nonculprit artery may be impaired in patients with STEMI. Although the perfusion of nonculprit artery may be improved after primary PCI in culprit artery, but it was still lower than those in the control group, and inflammation mechanism might contribute to it.