OBJECTIVE:To study the effects of podophyllotoxin derivative QW-83 on human cervical cancer HeLa cell apopto-sis and its mechanism. METHODS:After treated with 0(negative control),0.01,0.1,1 and 10 μmol/L QW-83 and positive drug etoposide(VP-16)for 48 h,proliferation inhibition rate and IC50 of HeLa cell were determined by MTT assay. The morphological changes of HeLa cell were observed by Hochest 33342 staining after treated with QW-83 [0(negative control),2.5,5,10μmol/L] for 48 h;flow cytometry was used to detect apoptosis rate;semi quantitative RT-PCR was adopted to detect the expression of apop-tosis related gene P53,Bax,Casepase-3,Casepase-8,Casepase-9 and Bcl-2 mRNA. RESULTS:Compared with negative control, 1,10 μmol/L VP-16 and QW-83 had obvious proliferation inhibition effect on HeLa cells (P<0.05 or P<0.01),and IC50 were (5.11±0.43)μmol/L and(4.96±0.54)μmol/L. Hochest 33342 staining results showed QW-83 could obviously induce cells apopto-sis and nuclear pyknosis. Flow cytometry showed QW-83 could increase apoptosis rate in concentration-dependent manner,being 16.89%-62.56%. RT-PCR showed mRNA expression of P53,Bax,Caspase-3,Casepase-8 and Casepase-9,Bcl-2/Bax increased, while mRNA expression of Bcl-2 decreased after treated with QW-83(P<0.05). CONCLUSIONS:Podophyllotoxin derivative QW-83 can induce HeLa cell apoptosis,and its mechanism may be associated with regulate mRNA expression of apoptosis related gene.

An investigation and analysis on the effect of current bilingual teaching has been done among the 2002rd seven-year program students by using questionnaire.The result can provide basic foundation for improving the quality of bilingual teaching.

Objective To monitor right heart hemodynamic changes of patients with lung cancer during and after the procedures of pneumonectomy and discuss the effect of pneumonectomy on right heart function and risk of postoperation morbidities. Method 16 patients with lung cancer were randomly selected and the clinical database was queried and Swan-Ganz catheter was applied awake from jugular vein to pulmonary artery ,measuring mean arterial pressure(MAP) ,heart rate(HR) ,central venous pressure(CVP) ,mean pulmonary artery pressure (MPAP),mean pulmonary capllary wedge pressure(MPCWP) ,and eardiaoc output(CO) and calculating cardiac index (CI),left ventricular-stroke work index (LVSWI), right ventricular stroke work index (RVSWI) , and stroke volume index(SVI) instantaneously before anaesthesia, after anaesthesia with single lung ventilation, after pulmonary artery and pulmonary vein occlusion and supine chest dosed during the procedure of pneumonectomy. Pre-and post-operative complications were recored. Results Cardiovascular complications occurred in 6 patients(37.5%) postoperatively. There were no operative or perioperative deaths. MPAP increased significantly after the procedure of pneumonectomy compared with that of preoperation, and MPAP > 26 mm Hg was in 4 patients who got cardiovascular complications postoperatively with morbidity of 100% ,which was "significantly higher than the morbidity of 16.7% (2/12) when MPAP < 25 mm Hg. Conclusion Pneumonectomy has significant effects on right heart hemodynemic changes and as MPAP increases postoperatively, the risk of cardiovascular complications becomes higher.

Objective To investigate the effects of 5-Aza-2′-deoxycytidine (5-Aza-Cde)on DNA methylation and expression of hMLH1 and MGMT gene in the human lung cancer cell line A549/DDP.Methods A549/DDP cells were cultured with RPMI 1 640 medium and were treated with 5 μmol/L DNA methyhransferase inhibitor 5-Aza-Cde.Methylation-specific pol-ymerase chain reaetioll (MSP)was used to detect the promoter methylation state of the hMLH1 and MGMT gene.RT-PCR was used to detect the mRNA expression of hMLH1 and MGMT before and after treatment with 5-Aza-Cde,respectively. Results Before treatment with 5-Aza-Cde,hMLH1 and MGMT expressions were absent,and promoter hypermethylation of the hMLH1 and MGMT gene were detected in A549 cells.After treatment with 5-Aza-Cde,the promoter region of the hM-LH1 and MGMT gene exhibited a demethylation state,and their mRNA expressions were increased.Conclusion Promoter hypermethyhtion is amajor mechanism of hMLH1 and MGMT gene silencing in human lung cancer cells,and can be reversed by the demethylating agent 5-Aza-Cde,which can regulate the expressions of the hMLH1 and MGMT gene.

Animals ; Hindlimb ; blood supply ; Intestine, Small ; metabolism ; Ischemia ; metabolism ; prevention & control ; Ischemic Preconditioning ; Lung Injury ; metabolism ; Male ; Muscle, Skeletal ; metabolism ; Rats ; Rats, Wistar ; Reperfusion Injury ; metabolism

Objective To investigate the effects of ischemic pre- and postconditioning on cerebral glycogen synthase kinase-3 beta (GSK-3β) activity in a rat model of global cerebral ischemia-reperfusion (I/R).Methods Forty male Wistar rats weighing 200-230 g were randomly allocated into 4 groups (n =10 each) : Ⅰ group sham operation (group S); Ⅱ group I/R; Ⅲ group ischemic preconditioning (group IPR) and Ⅳ group ischemic postconditioning (group IPO). The animals were anesthetized with intraperitoneal 10% chloral hydrate 0.4 ml/100 g. Global cerebral ischemia was induced by four-vessel-occlusion in group Ⅱ , Ⅲ and Ⅳ. Bilateral vertebral arteries were cauterized and bilateral carotid arteries were occluded for 10 min. In group IPR cerebral ischemia was preceded by 3 cycles of 10 s ischemia followed by 30 s reperfusion. The group IPO received 3 cycles of 30 s reperfusion followed by 10 s ischemia at the end of 10 min cerebral ischemia. The animals were killed 2 days later. The brains were immediately removed for determination of neuronal apoptosis in the cortex (by TUNEL), the infarct size (by TTC), p-GSK-3β activity (by spectrum assay) and the expression of Bcl-2, Bax and Caspase-3 (by SP). Linear correlation of p-GSK-3β activity with the number of apoptotic neurons in the cortex and cerebral infarct size was analyzed. Results Cerebral I/R significantly increased the number of apoptotic neurons in the cortex and infarct size, decreased p-GSK-3β activity, down-regulated Bcl-2 expression and up-regulated Bax and Caspase-3 expression in group I/R as compared with group S. Ischemic pre- and postconditioning significantly attenuated these cerebral I/R-induced changes. The p-GSK-3β activity was negatively correlated with the number of apoptotic neurons in the cortex and cerebral infarct size. Conclusion Ischemic pre- and postconditioning reduces cerebral I/R injury through inhibiting the activity of GSK-3β.

Current clinical evaluation of literature quality has various ways. Most of them lay special emphasis on the evaluation of the design quality, but the evaluation of the implementation process quality is not perfect. Especially data management is not fully emphasized during the enforcement of clinical trials. Data from clinical research were bases for evaluating clinical findings. Although strict specifications and requirements for data management might be strictly written clearly in research protocols, they were not embodied in current clinical research evidence evaluation system. Data management is an important part of implementing the whole clinical trial process, which is a comprehensive reflection of data collecting, logging, sorting, and managing. Its objective is to obtain high quality research data for statistical analysis, thereby coming to a true and reliable conclusion. In order to overall evaluating clinical design and implement, we suggest that present quality evaluation indicators of clinical trails should be completed, and add data management quality evaluation during the whole implement process. Data management plans, standards and requirements for data checking, and management regulations for disobeying data and exception data should be added in quality evaluation indicators for clinical research evidence. The effect of data management quality on clinical research evidence evaluation should be emphasized.
Biomedical Research ; Humans ; Research Design

OBJECTIVETo observe the clinical effect of Guoshu acupoint pressure therapy on acute mastitis during lactation.

METHODSFifteen cases suffered from acute lactation mastitis were treated with Guoshu acupoint pressure therapy, that is, firstly with lifting and flicking reduction at "Taiji" and "Xuepen" point, whose intensity was varied from patient's physical fitness. Subsequently, the patients were treated with flame therapy induced by distillate spirit, once each day.

RESULTSAfter the treatment, all the patients were cured completely in from 1 to 5 days, with an average of 2.5 days.

CONCLUSIONGuoshu acupoint pressure therapy is effective on acute mastitis during lactation.

Aim To study the mechanism of action of the new derivative of podophyllotoxin(LN-13)in indu-cing the apoptosis of K562/A02 cells.Methods The MTT method was taken to detect the inhibition of LN-13 and VP-16 on K562/A02 proliferation and inhibi-tion rate and IC50 values were obtained 48 hours later. The K562/A02 cell morphological change induced by LN-13 were observed through Hochest33342 and PI staining after 48 hours later.Flow cytometry was taken to detect the apoptosis of K562/A02 cells induced by LN-13.The reverse transcription-polymerase chain re-action was taken to detect the Bcl-2,Bax,caspase-3 and mdr-1 mRNA expression.The expression of P-gp was detected by Western blot.Results The growth of K562 /A02 cells was obviously inhibited by LN-13 when IC50 value was 3.32 μmol · L-1 .LN-13 could obviously induced cell apoptosis observed by Ho-chest33342 and PI staining.Flow cytometry detection showed that LN-13(2,4,8 μmol·L-1 )could induce cell apoptosis and apoptosis ratio reached 15.0%, 48.0%,68.96%,respectively.The reverse transcrip-tion-polymerase chain reaction showed that LN-13 in-creased the Bax and Caspase-3 mRNA expression,and meanwhile the expression of mdr-1 mRNA decreased. Western blot showed that P-gp expression was de-creased as the LN-13 dose increased.The data were significantly different from those of control group.Con-clusion Podophyllotoxin derivative LN-13 can induce the apoptosis of K562 /A02 cells,which may be close-ly-related to regulating P-gp expression and apoptosis related gene mRNA expression.

Objective To evaluate the application value of bedside lung ultrasound in diagnosis of neonatal severe pneumonia.Methods Bedside lung ultrasound and chest X-ray were performed in 59 the newborn patients admitted to the newborn department for suspected severe pneumonia,from October 2015 to March 2016.According to the characteristics of the chest X-ray,patients were divided into consolidation group (widely spot shadow in double lung and airbronchograms) and the unconsolidation group (enlargement,fuzzy texture in double lung or small patch sample changes) Differences in ultrasonic characteristics were compared between the two groups.Results Ultrasonic image characteristics were pulmonary consolidation with air bronchograms,A-line disappearance,pleural line abnormalities.Among 23 patients with chest X-ray,21 patients were found pulmonary consolidation by lung ultrasound,and all 36 patients in unconsolidation group were not found pulmonary consolidation by lung ultrasound,and there was no significant difference in the detection rate by lung ultrasound between the two groups (P＞0.05) The pulmonary ultrasound results of 20 cases in the consolidation group showed A-line disappearance,37 cases in the unconsolidation group showed A-line disappearance.There was no statistical significant differences between the two groups (P＞0.05).The pulmonary ultrasound results showed 11 cases in the consolidation group and 15 cases in the unconsolidation group with pleural line abnormalities,there was no statistically significant differences between the two groups (P＞O.05).Conclusions Ultrasound image characteristics of neonatal severe pneumonia include pulmonary consolidation and air bronchograms,A-line disappearance,pleural line abnormalities.Compared lung ultrasound and chest X-ray result,there was no difference between consolidation group and unconsolidation group.Both of the two methods can be used for evaluating the diagnosis of pneumonia.