Objective HPLC method was used to determine strychnine and brucine in the decoction of Semen Strychni and its processed products of Jiangxi methods and innovated methods.Methods SiO2 was used as the stationary phase,n-hexane∶dichloromethane∶methanol∶ammonia(47.5∶47.5∶5∶0.35) as the mobile phase,with detection wavelength at 254 nm.Result The contents of strychnine and brucine in the processed products of Jiangxi methods and innovated methods were low.The content of that in acetic acid-soaked product scalded with hot sand was the lowest.Conclusion Semen Strychni and its processed products of Jiangxi methods and innovated methods can lower the toxicity and improve the safety of clinical use.

Objective To observe the rat cardiac size and cardiac function changes before and after trimetazidine administration plus bone-marrow stem cells transplanting through echocardiography.Methods Forty wistar rats were divided into the following 4 groups randomly:control group (T),myocardial infarction group (Ⅱ),bone marrow stem calls transplantation group (Ⅲ),and bone marrow stem cells transplantation plus trimetazidine administration group(Ⅳ).The rats' left anterior coronary artery in group Ⅱ,Ⅲ and Ⅳwas ligated to produce myocardial infarction model,then bone-marrow stem cells were injected around the infarcted area into the later two groups.Furthermore,rats in group Ⅳ were administrated with trimetazidine.The size and systolic function of the hearts were measured 4 weeks after transplantation.The left ventricular systolic pressure(LVSP) and the end-diastolic pressure(LVEDP) were also measured at the end of experiment.Results The left ventricular diameter of rats in group Ⅲ and Ⅳ was smaller than that in group Ⅱ,and the ventricular systolic function increased,LVSP increased and LVEDP decreased statistically in group Ⅲ and Ⅳ.the amelioration of cardiac size and function was significantly notable in group Ⅳ than that in group Ⅲ.Conclusions Bone marrow stem cells transplantation can release the enlargement of left ventricle and improve cardiac function after myocardial infarction.The therapeutic efficacy can be further elevated if administrated with trimetazidine simultaneously.

Objective To discuss the clinical application of homemade airway exchange catheter on the extubation of patients with difficult tracheal intubation in intensive care unit(ICU). Methods Sixty-two patients with difficult tracheal intubation who failed their initial extubation trial were randomly divided into conventional group (31 cases)and observation group(31 cases). The patients in the conventional group received routine extubation process,while the patients in the observation group were extubated under the guidance over a homemade airway exchange catheter. The changes in heart rate(HR),blood pressure,respiratory rate(RR)and pulse blood oxygen saturation(SpO2)were compared at 12 hours after extubation,so as the re-intubation rate,intubation success rate at first attempt and re-intubation time in two groups,and the tolerance and complications after extubation were observed. Results After extubation,the HR,blood pressure and RR were increased significantly(all P<0.05), and the SpO2 was much lower in conventional group(P<0.05),while those parameters were changed little and basically in the normal ranges in the observation group. At 12 hours after extubation,the re-intubation rate was much lower(6.45%vs. 25.81%,P<0.05)in the observation group,with shorter re-intubation time(seconds:27±14 vs. 49±28,P<0.01),higher intubation success rate at first attempt(90.32%vs. 54.84%,P<0.01)and better tolerance (77.4% vs. 61.3%,P<0.05)compared with those in the conventional group. There was no severe complication in the observation group,and there were 1 cases of glottic edema with cricothyroid membrane puncture,2 cases of broncheal mucous membrane bleeding and 2 cases of bucking in the conventional group. Conclusion Compared with conventional extubation process,the extubation over homemade airway exchange catheter can increase the rate of extubation,reduce re-intubation rate and the re-intubation time,with favorable tolerance and no occurrence of serious complications,and is one of the safe and effective extubation strategies in patients with difficult tracheal intubation in ICU.

Twenty-one rabbits were randomly divided into three groups: experimental group-filling vein with two neural segments being 0.3cm long to bridge 4cm defect of common peroneal nerve, control group-bridging vein directly to the 4cm defect of common peroneal nerve, auto-nerve-grafting group-cutting off a segment (4cm long) from common peroneal nerve and grafting it inversely.After 25 weeks, morphological, electrophysiological and histological examinations were undertaken, which revealed that the experimental group was most similar to the auto-nerve-grafting group in recovery of motion of the limbs, action potential of muscle, nerve conductive velocity, and regenerating density of nerve fibers and axons. It was a failure in control group. Satisfactory results were also achieved in 2 patients with defect of ulnar nerve treated by filling vein with two neural segments. It suggests that our method is feasible.

Objective To evaluate the safety and effectiveness of a curved vertebroplasty (CVP) compared with traditional unipedicular approach vertebroplasty (UVP) in treating osteoporotic vertebral compression fractures (OVCF).Methods This was a retrospective case control study on the clinical data of 77 OVCF patients (12 males,65 females;aged 55-86 years,mean 70.8 years) admitted between July 2013 and December 2016.There were 6 injured vertebrae at T1 10,73 at T11 L2,and 12 at L3 5.The patients were divided into CVP group (36 patients,44 vertebrae) and UVP group (41 patients,47 vertebrae) with no significant difference in baseline clinical variables.Intraoperative and postoperative complications including neurovascular injury were recorded.Operation duration,fluoroscopy frequency,volume of cement per level,cement leakage rate per level treated,cement distribution,and refracture rate were compared between the two groups.Preoperative and postoperative visual analog scale (VAS) and Oswestry disability index (ODI) were compared both within the group and between the groups.Results No severe complications related to puncture were observed.No significant difference was observed for operation duration,fluoroscopy frequency,and cement leakage rate per level treated between the two groups (P ＞ 0.05).Compared with UVP group,CVP group had larger volume of cement per level [(5.0 ± 1.4) ml vs.(4.3 ± 1.6) ml],more uniform cement distribution (none vs.10 cases),and lower refracture rate (0 vs.10％) (P ＜ 0.05).The two groups were followed up for 6-49 months (mean,25.9 months).Significant improvements on the VAS and ODI were noted within each group (P ＜0.01),but there was no significant difference between the two groups (P ＞ 0.05).Conclusions Both CVP and UVP are safe and effective treatments for OVCF.Compared with UVP,CVP entails more uniform cement distribution and lower refracture rate.

Objective:To investigate the relationship between YES-related protein 1(YAP1)and prostate-specific antigen(PSA)in human castration-resistant prostate cancer(CRPC), and explore the regulation mechanism of YAP1 on PSA.Methods:The luciferase reporter gene was used to detect the activity change of the PSA gene promoter region after the over expression of YAP1 in LNCaP and C4-2 cells.The effect of over expression of YAP1 gene on PSA protein in different prostate cancer cell lines was detected by Western blot(WB)method, and the effect of YAP1 silencing on PSA protein in C4-2 cells was observed.The Q-PCR method was used to further verify the expression change of PSA mRNA affected by YAP1 gene over expressed in C4-2 cells.Meanwhile, WB was used to explore the effect of YAP1 on androgen receptor(AR)in C4-2 cells.Results:After over expression YAP1 in CRPC, the luciferase experiment showed that the average C4-2 cell ratio of experimental group to control group was 3.17815892(>2 times, P<0.001). After Q-PCR detection of all over-expressed YAP1 gene fragments, the measured PSA mRNA values in the experimental groups were 2.306667, 1.553333333, 2.613333333, and 2.673333333, respectively, which were higher than those in the control group(1 time, P<0.001), indicating that the PSA expression was significantly increased.WB analysis showed that after C4-2 cells over expressed YAP1, the AR band was significantly enhanced in the experimental group compared with the control group, suggesting that the AR protein expression in the nucleus was significantly increased in the YAP1 over expression group. Conclusions:YAP1 might positively regulate the PSA expression in CRPC and have an ability to promote AR translocation into the nucleus.

Objective: To investigate the change of autophagy level of bone marrow nucleated red blood cell (RBC) in patients with myelodysplastic syndromes (MDS) . Methods: Fifty-four MDS patients and thirty-three controls were enrolled in this study. The mitophagy were observed by transmission electron microscopy (TEM) . The level of autophagy-associated protein LC3B in GlycoA⁺ nucleated RBC was measured by flow cytometry. The expressions of ULK1 and mTOR mRNA in GlycoA⁺ nucleated RBC were measured by real-time PCR. The expression of the mitochondrial outer membrane protein TOM20 in GlycoA⁺ nucleated RBC was detected by Western blot. Results: Autophagosomes or autolysosomes were scarcely observed by TEM in MDS patients. The expression of LC3B in GlycoA⁺ nucleated RBC in high-risk MDS patients (0.22±0.12) was significantly lower than that in normal controls (0.43±0.22, P<0.001) , and lower than that in low-risk MDS patients (0.40±0.16, P=0.001) . The expression of AMPK [0.26 (0.60) ] in GlycoA⁺ nucleated RBC in high-risk MDS patients was significantly lower than that in controls [1.00 (2.07) , P<0.017) . The expression of ULK1 mRNA in GlycoA⁺ nucleated RBC in high-risk MDS patients [0.27 (3.31) ] was significantly lower than that in controls [1.07 (4.41) , P<0.017]. The level of mTOR mRNA in GlycoA⁺ nucleated RBC in high-risk MDS patients [1.82 (3.74) ] was significantly higher than that in controls [1.26 (1.38) , P<0.017]. The level of LC3B in GlycoA⁺ nucleated RBC was negatively correlated with the HGB (r=0.529, P=0.009) in high-risk MDS patients. The expression of mitochondrial outer membrane protein TOM20 in high-risk MDS patients was 9.42±4.42. Conclusion Autophagy is impaired in nucleated RBC of MDS patients.

Objective: To explore characteristic and function of peripheral blood mononuclear cells (PBMNC) -induced macrophages in patients with myelodysplastic syndrome (MDS) to couple with its progression. Methods: A total of 24 MDS patients (11 low-risk patients and 13 high-risk group patients) referred to Department of Hematology of Tianjin Medical University General Hospital and normal controls were enrolled from September 2014 to December 2015. PBMNC was stimulated with GM-CSF to transform to macrophages. The morphology of macrophages was observed by microscope. The quantity of macrophages, CD206 and SIRPα on surface of macrophages were detected by flow cytometry. The phagocytic function of macrophages was analyzed by fluorescence microscopy and flow cytometry. Results: The morphology of macrophages from MDS patients was abnormal. The percentage of transformed macrophages was (5.17±3.47) % in patients with MDS, which was lower than that in controls significantly[ (66.18±13.43) %, t=3.529, P=0.001]. The expression of CD206 on macrophages from MDS patients was significantly lower than that of controls[ (9.73±2.59) % vs (51.15±10.82) %, t=4.551, P<0.001]. The SIRPα level of macrophages from MDS patients was significantly lower than that of controls [ (0.51±0.09) % vs (0.77±0.06) %, t=2.102, P=0.043]. The phagocytic index and the percentage of phagocytic of macrophages from MDS patients were significantly lower than those of macrophages from normal controls[0.45±0.08 vs 0.92±0.07, t=-6.253, P=0.008; (23.69±3.22) % vs (42.75±2.13) %, t=-6.982, P=0.006 respectively]by flow cytometry. The phagocytic index of MDS patients was significantly lower than that of controls (0.24±0.04 vs 0.48±0.96, t=3.464, P=0.001) by fluorescence microscopy. Conclusion The quantity, recognization receptors and phagocytosis of PBMNC-induced macrophages decreased in MDS patients.

OBJECTIVETo investigate the change of autophagy level of bone marrow mononuclear cells（BMMNCs）in patients with myelodysplastic syndromes（MDS）.

METHODSThirty- eight patients with MDS and 26 megaloblastic anemia patients were enrolled in this study. The autophagic vacuoles were observed by transmission electron microscopy （TEM） and the quantity of autophagic vacuoles was detected by monodansylcadaverine （MDC） staining. The LC3 protein positive cells were counted by immunofluorescence assays. The expression of Beclin 1, LC3A, mTOR mRNA were measured by real time PCR. The expression of Beclin 1 proteins were detected by Western blotting.

RESULTSThe autophgic vacuoles of double membrane that surrounds lysosomes appeared in MDS patients. The percentage of MDC positive cells was significantly higher in MDS patients［（9.75±2.63）%］than that of controls［（2.90± 0.89）%, P＜0.05）. The percentage of LC3 protein cells was also increased in MDS patients（6.13±1.03）% vs（1.5±0.58）%, P＜0.05）. The expression of Beclin 1 and LC3A mRNA in low-risk and intermediate-1 MDS were higher compared with controls （3.61 ± 3.02 vs 1.55 ± 1.03 and 6.56 ± 3.97 vs 1.21 ± 0.95 respectively, both P＜0.05）. The expression of mTOR mRNA was down- regulated in low- risk and intermediate-1 MDS compared with controls（0.39±0.37 vs 1.50±1.03, P＜0.05）. There were no significant difference in expression of Beclin 1, LC3 and mTOR mRNA among intermediate-2 and high-risk MDS and controls. Beclin 1 protein expression was higher in low- risk and intermediate- 1 MDS patients（1.257 ± 0.197）than that of controls（0.528±0.086）and inermediate-2 and high-risk MDS patients（0.622±0.118）.

CONCLUSIONThe autophagy levels were increased in low- risk and intermediate- 1 MDS, while not enhanced in intermediate-2 MDS. Autophagy might be considered as a cell protective mechanism in MDS. The relatively defective autophagy in intermediate- 2 and high- risk MDS might contribute to disease's progression.

Apoptosis Regulatory Proteins ; metabolism ; Autophagy ; Beclin-1 ; Bone Marrow Cells ; cytology ; Humans ; Membrane Proteins ; metabolism ; Microscopy, Electron, Transmission ; Microtubule-Associated Proteins ; metabolism ; Myelodysplastic Syndromes ; pathology ; TOR Serine-Threonine Kinases ; metabolism ; Vacuoles ; ultrastructure