Based on the teaching concept of constructivism, this study aims to promote independent inquiry-based learning and clinical thinking among students and establish the guiding ideology of "full participation, process control, in-depth discussion, and expansion of thinking". A blending learning model was adopted with offline inquiry-based group learning and in-class defense and comment, as well as online teacher-student interaction and supervision to promote learning. Case-problem-based learning (CPBL) of pathophysiology was carried out among the medical students in the class of 2017, and process management was strengthened to effectively manage the two key links of data retrieval and group discussion. The analysis of 176 teaching evaluations collected at the end of the semester show that in terms of the overall evaluation of CPBL teaching, 162 students (92.05%) had high evaluation on teaching objectives, organization, cases, and personal gains and held a very or relatively favorable attitude. There were more negative feedbacks on "appropriate time allocation"; 21 students (11.93%) held a relatively or very disapproving attitude, and 149 students (84.66%) "felt very tired". In terms of teaching effect evaluation, 150 students (85.23%) strongly or relatively agreed that CPBL teaching may help to understand professional knowledge, stimulate learning enthusiasm and initiative, improve problem solving ability, emphasize clinical practice to cultivate clinical thinking, supervise and promote learning, and enhance team cooperation and teacher-student communication. In terms of the evaluation of teachers, 167 students (94.89%) thought that teachers were rigorous, responsible, and enthusiastic in teaching, attached importance to process management, and did well in effective guidance and thinking inspiration (strongly or relatively agree). The above results suggest that the CPBL teaching reform of pathophysiology based on process management can effectively promote in-depth inquiry-based independent learning and the cultivation of clinical thinking and improve teaching effectiveness, but further improvement is needed for teaching arrangement and time allocation.

Objective:To analyze the clinical characteristics of patients with acute glyphosate herbicide poisoning and the differences in the severity of poisoning.Methods:A retrospective analysis was performed on patients with acute glyphosate herbicide poisoning admitted to the First Affiliated Hospital of Zhengzhou University from January 2014 to December 2020. The general information, exposure time, poisoning dose, poisoning cause, poisoning route, clinical manifestations, laboratory examination results during hospitalization, treatment measures, hospital stays and prognosis of the patients were collected. The patients were graded according to the poisoning severity scoring standard of Chinese Expert Consensus on Diagnosis and Treatment of Acute Poisoning in 2016. The highest severity score during hospitalization was used as the final grade. According to the final grade, asymptomatic and mild patients were included in the mild group, and moderate, severe and death patients were included in the severe group. The independent sample T test or Mann-Whitney U test was used for measurement data, and χ2 test or Fisher's exact test was used for counting data. The differences of general data and clinical data between the two groups were compared. Results:According to the inclusion and exclusion criteria, 83 patients with acute glyphosate herbicide poisoning were selected as the study subjects. All patients survived, mainly mild poisoning (56.6%), with a male to female ratio of 33∶50, and an average age of 39 years. The number of poisoning cases increased yearly (the highest in 2019), and most cases occurred in spring and summer. The main cause of poisoning was suicide (71.1%), direct oral administration (83.1%) was the primary route of poisoning, and the dominating clinical manifestations were digestive symptoms (71.1%). Laboratory tests showed increased white blood cell count (WBC), neutrophil percentage (NEUT %) and D-dimer, and decreased hemoglobin and potassium. Compared with the mild group, patients in the severe group were older [(51±17) years vs. (35±19) years], had a higher proportion of suicide and direct oral administration, a longer hospital stay [8.0 (4.8, 12.0) d vs. 3.0 (2.0, 5.5) d], a higher dose of poisoning [200.0 (50.0, 200.0) mL vs. 30.0 (11.3, 57.5) mL], and higher NEUT % within 24 h of admission [(83.4±10.4) vs. (73.2±12.8)]. The increase of WBC, NEUT %, aspartate aminotransferase, prothrombin time, D-dimer and the decrease of serum potassium were more common in the severe group than the mild group, with statistical significance (all P<0.05). Conclusions:The number of patients with acute glyphosate herbicide poisoning is increasing yearly. Generally, the condition is mild and the prognosis is satisfying. The severity is more serious in the middle-aged and elderly patients andthose with direct oral administration, high toxic dose, and high NEUT % within 24 h of admission. Severe poisoning is more likely to cause changes in laboratory indicators.

Objective To determine ED50 and ED95 of oxycodone inhibiting responses to endotracheal intubation together with target concentration infusion of propofol.Methods ASA physical status Ⅰ or Ⅱ patients,aged 18-65 years,with body mass index 18.5-24.9 kg/m2,falling into Mallampati Score Ⅰ or Ⅱ,undergoing elective surgery under general anesthesia,were enrolled.Anesthesia was induced with intravenous injection of oxycodone,target concentration infusion of propofol with plasma concentration of 4μg/ml and intravenous injection of rocuronium 0.9 mg/kg when BIS＜60.The patients were endotracheally intubated and mechanically ventilated.The initial dose of oxycodone was 0.2 mg/kg.The dose of oxycodone was increased/decreased by ratio of 1.1 in the next patient by modified Dixon's up-and-down method.The response to endotracheal intubation was defined as an increase in the maximum mean arterial pressure and/or the maxmium heart rate≥20％ of the baseline value with 2 minute after intubatior.Probit analysis was used for calculating ED50,ED95 and 95％ confidence interval (CI) of oxycodone inhibiting responses to endotracheal intubation.Results There were twenty-seven patients who completed the test finally.ED50 and ED95 (95％ CI)of oxycodone inhibiting responses to endotracheal intubation were respectively 0.204 (0.175-0.249)mg/kg and 0.342(0.287-0.409) mg/kg.Conclusion When plasma target concentration of propofol 4 μg/ml is infused,ED50 and ED95 of oxycodone inhibiting responses to endotracheal intubation is 0.204 and 0.342 mg/kg,respectively.

Objective To study the effect of Bairui granule and Deacetylated Ligustrazin injection on plasma BNP and FABP levels in children with heart failure and pneumonia.Methods 60 children of heart failure and pneumonia who received therapy from July 2014 to June 2016 in our hospital were randomly divided into two groups, the observation group and the control group, 30 cases in each group.children in the observation group were treated with sodium phosphate creatine, and the control group was treated with 2-linolenic acid,the clinical outcomes, clinical signs, BNF, FABP, LVEF and LVEDP were compared between the two groups.Results After treatment, the total effective rate of the observation group was significantly higher than that of the control group, the difference was statistically significant (P<0.05); The heart rate, respiratory rate and blood oxygen saturation of the two groups were significantly improved compared with those before treatment ( P<0.05 ) , the heart rate and respiratory rate of the observation group were lower than those of the control group, the oxygen saturation was higher than that of the control group (P<0.05).After treatment, the BNF and FABP indexes of the two groups were lower (P<0.05), and the BNF and FABP indexes in the observation group were lower than those in the control group (P<0.05); The LVEF of the observation group was higher than that of the control group, the LVEDP index was lower than that of the control group, the difference was statistically significant (P<0.05).Conclusion Bairui granule and Deacetylated Ligustrazin injection in the treatment of heart failure with pneumonia has a good effect in the process, which will help reduce the BNP index in children, recovery of children's body function.

PURPOSE: The aim of our study was to explore the relationships between the M2 isoform of pyruvate kinase (PKM2) and the sensitivity of human non-small cell lung cancer (NSCLC) cells to docetaxel in vitro. MATERIALS AND METHODS: With the method of plasmid transfection, we silenced the expression of PKM2 successfully in A549 and H460 cells. Western blotting and real-time PCR were applied to detect PKM2 expression at protein and gene levels. Cell viability was examined by CCK8 assay. Cell cycle distribution and apoptosis were examined by flow cytometry. P21 and Bax were detected. RESULTS: Expression of PKM2 mRNA and protein were significantly decreased by shRNA targeting PKM2. Silencing of PKM2 increased docetaxel sensitivity of human NSCLC A549 and H460 cells in a collaborative manner, resulting in strong suppression of cell viability. The results of flow cytometric assays suggested that knockdown of PKM2 or docetaxel treatment, whether used singly or in combination, blocked the cells in the G2/M phase, which is in consistent with the effect of the two on the expression of p21. Cells with PKM2 silencing were more likely to be induced into apoptosis by docetaxel although knockdown of PKM2 alone can't induce apoptosis significantly, which is in consistent with the effect of the two on Bax expression. CONCLUSION: The results suggest that PKM2 knockdown could serve as a chemosensitizer to docetaxel in non-small lung cancer cells through targeting PKM2, leading to inhibition of cell viability, increase of cell arrest of G2/M phase and apoptosis.
Apoptosis ; Blotting, Western ; Carcinoma, Non-Small-Cell Lung ; Cell Cycle ; Cell Line* ; Cell Survival ; Drug Therapy ; Flow Cytometry ; Humans* ; In Vitro Techniques* ; Lung Neoplasms ; Methods ; Plasmids ; Pyruvate Kinase* ; Pyruvic Acid* ; Real-Time Polymerase Chain Reaction ; RNA, Messenger ; RNA, Small Interfering* ; Transfection

BACKGROUND:Brahma-related gene 1 (Brg1), a catalytic subunit of an important chromatin remodeling complex, has been considered as a key nuclear transcriptional factor, and tends to be decreased in diabetic cardiomyopathy.
OBJECTIVE:To construct an adenovirus vector carrying Brg1, and observe its protective role in oxidative stress induced-cardiomyocyte apoptosis.
METHODS:The recombinant adenovirus plasmid was linearized and transfected into HEK293 cel s using Fugene HD for packaging and amplification. The adenovirus particles were further purified, quantified, and sequential y transfected to cardiomyocytes of neonatal Sprague-Dawley rats. The Adeno-EGFP transfected and non-transfected cardiomyocytes were used as control group. 24 hours later, the transfection efficiency was observed by fluorescent microscope, and expressions of Brg1 mRNA and protein were detected by quantified PCR and western blotting. After treatment with 100 μmol/L H2O2 for 12 hours, the expressions of Brg1 protein and cleaved-Caspase 3 were measured by western blotting, and cel apoptosis was analyzed by flow cytometry.
RESULTS AND CONCLUSION:(1) The recombinant adenovirus vector of Brg1 had been successful y transfected into cardiomyocytes with higher expressions of Brg1 mRNA and protein, and the transfection efficiency reached more than 90%. (2) After H2O2 treatment, the Brg1 was significantly down-regulated in contrast to the up-regulation of cleaved-Caspase 3;the flow cytometry data showed that the apoptotic cel s were increased. But in Adeno-Brg1 transfected cardiomyocytes, the H2O2 induced cel apoptosis was significantly decreased compared with non-transfected cel s and empty vector transfected cel s. (3) These results suggest that oxidative stress can directly inhibit the Brg1 expression, and overexpression of Brg1 can protect the cardiomyocytes from cel apoptosis induced by oxidative stress.

AIM:To probe whether CpG oligodeoxyribonucleotides 7909 (CpG ODN7909) combined with Toll like receptor (TLR)9 affected the chemotherapeutic sensitivity of doctaxel (DOC) in human lung cancer A549 and H520 cell lines.METHODS:Sequences of TLR9 siRNAs were designed.A549 and H520 cells were transfected with TLR9 siR-NA by lipofectamine.The expression of TLR9 was detected by Western blot .The cell activity was measured by CCK-8 as-say.The experiments were divided into blank control group , control siRNA group and TLR9 siRNA interference group.The cell cycle distribution and cell apoptosis were analyzed by flow cytometry .The expression of P38 and Bax was determined by Western blot .The cells in each group were exposed to CpG ODN 7909 and/or DOC.RESULTS: In A549 cells and H520 cells, CpG ODN7909 alone had no obvious effect on the cell activity , G2/M phase arrest and apoptosis , but in-creased the protein expression of P 38 and Bax ( P<0.01) .In addition, there was no significant changes of the above inde-xes in CpG ODN7909 treated-TLR9 siRNA group was observed .DOC alone significantly inhibited the cell activity , higher the G2/M phase fractions, apoptotic rates and Bax expression (P<0.01), but didn’t affect the expression of P38 in all 3 groups.Compared with the cells treated with DOC alone , the cells treated with CpG ODN7909 combined with DOC exhibi-ted lower cell activity, higher G2/M phase fractions, apoptosis rates and more Bax expression (P<0.01), showed no sig-nificant change of P38 expression.In addition, there was no significant change of the above indexes in CpG ODN 7909 com-bined with DOC treated-TLR9 siRNA group was observed .CONCLUSION:CpG ODN7909 may enhance the chemothera-peutic sensitivity of DOC in human lung cancer cells by combining with TLR 9.The mechanism might be related to enhan-cing the inhibitory effect and apoptosis of DOC on the cell activity in vitro, arresting the cells at G 2/M phase of the cells .

Objective To observe the effect of Oxycontin combined with Gabapentin for treatment of malignant neuropathic pain.Methods Sixty-three cases of malignant neuropathic pain in Jinshan Hospital Affiliated to Fudan University were randomly divided into group A, B and C.Patients of which were given Oxycontin, Gabapentin, Oxycontin combined with Gabapentin respectively for pain treatment.The analgesic effects, toxic reaction side effects, quality of life, and immune function were all compared in three groups.Results Compared with pretherapy, the cancer pain score (NRS), quality of life (QOL) and karnofsky performance status(KPS) scores in all groups were changed significantly after drugs therapy(F=375.852,154.612, 151.838,P＜0.05).The levels of CD3,CD4, CD4/CD8 and NK cells in all groups were higher than before therapy(F=158.935,108.145,366.973,92.090,P＜0.05).After treatment,the NRS, QOL and KPS scores in group C were 2.00± 0.86,44.80± 6.07, 84.50± 6.05, in group A were 3.35 ± 0.67,37.35 ± 5.71,74.50 ±10.99,and in group B were 4.05±0.94,35.85±5.90,72.00±8.34, and the different were significant (F =3.250,10.499,3.465,P＜0.05).The levels of CD3, CD4, CD4/CD8and NK cells in group C were (72.94 ±5.63)％,(41.52±4.19)％, 1.86±0.30, (27.57±6.86)％,in group A were (62.84±5.27)％, (33.84 ±5.40)％,1.35±0.37, (20.49±6.67) ％,and in group B were (62.22±8.10)％, (33.19±6.90)％, 1.32 ± ±0.41, (20.32±5.63) ％, and the different were significant (F =3.377,3.344,3.352,3.386, P＜ 0.05).The patient in group C had less adverse effects than those in group A and B.Conclusion Oxycontin and Gabapentin in treatment of malignant neuropathic pain is effective.

Objective To explore the role of pyruvate kinase M2 (PKM2) siRNA in the radiosensitivity of human lung cancer A549 cells.Methods PKM2 siRNA was synthesized according to the coding sequence of PKM2 mRNA and then was transferred into A549 cells with lipofectamine.The expressions of PKM2 gene and protein was detected by RT-PCR and Western blot,respectively.The experiments were divided into PKM2 siRNA interference group,siRNA negative control group,and blank control group.The cells of each group were exposure to 6 MV X-rays in different dose.Radiosensitivity was evaluated by colony formation assay.Flow cytometry was applied to analyze cell cycle distribution and apoptosis.Data are representative of three independent experiments.Results Ccompared with blank control cells,the expressions of PKM2 gene and protein in the PKM2 siRNA transferred A549 cell was efficiently diminished (t =20.91,47.00,P ＜0.01) with inhibition rates of (70.27 ± 1.38)％ and (70.42 ± 1.18) ％,respectively.Compared with control,PKM2 siRNA transfection significantly decreased the D0,Dq,N and SF2 values (t =43.82,28.44,15.60,29.63,P ＜ 0.01) and hence yield a sensitization enhancement ratio (SER) of 1.27.In addition,the percentage of G2/M phase cells in the siRNA group and irradiated group were both significantly higher than that of the blank control group (t =8.35,27.87,P ＜ 0.01).The combined treatments of PKM2 siRNA interference and irradiation arrested more cells in the G2/M phase compared to either treatment alone.The apoptosis rate of siRNA group was not dramatically different from that of blank control group.The apoptosis rate of irradiation group was higher than that of blank control group (t =23.99,P ＜ 0.01),and the combined treatments of siRNA and irradiation enhanced the apoptotic rate compared to either treatment alone (t=9.42,65.21,P ＜ 0.01).Conclusions Specific blockage of PKM2 expression by gene silencing could enhance the sensitivity of human lung cancer A549 cells to radiotherapy in vitro,which may due to the cell cycle arrest and apoptosis induction after irradiation.

Objective To investigate the relationship between the effect of unmethylated cytosine phosphate-guanine oligodeoxynucleotide ( CpG ODN ) 7909 on X-ray-induced G2/M phase arrest and apoptosis and the phosphorylation of ATM kinase.Methods Human lung adenocarcinoma A549 cells were randomly classified into five groups,control,CpG group,irradiation group,CpG + irradiation group and ATM siRNA + CpG + irradiation group.Cell survival fraction was evaluated by clonogenic assay.Cell cycle and apoptosis were analyzed by flow cytometry.The expressions of ATM,checkpoint kinase-2(Chk2) and p53 were detected with Western blot.Results Compared to the situation under X-ray irradiation alone,decreased cell clonogenic survival,prolonged G2/M arrest,and increased cell apoptosis were observed after the combination treatment with CpG ODN7909 and X-rays ( t =13.41,17.32 and 7.71,P ＜ 0.05).Moreover,the phosphorylations on ATM,Chk2,and p53 were increased in the irradiated A549 cells that had been pre-treated with CpG ODN7909.After ATM siRNA interfering,abrogation of G2/M arrest,reduction of apoptosis,and decrease of Chk2 and p53 phosphorylation were found in A549 cells treated with CpG ODN7909 and X-rays ( t =26.84,2.98,47.24 and 67.47,P ＜0.05).Conclusions CpG ODN7909 can enhance the X-ray-induced phosphorylation of ATM kinase in human lung adenocarcinoma cells in vitro,which might be involved in regulating G2/M phase arrest and apoptosis.