OBJECTIVETo investigate the apoptosis and proliferation effect of matrine on human medulloblastoma cell line D341 in vitro and the effect of the expression of the related caspase 3 and caspase 9 proteins.

METHODSThe D341 cells were cultivated successfully in vitro. Then the cells were divided into 5 groups according to the concentration of matrine (0.5 mg/mI group, 1.0 mg/ml group, 1.5 mg/ml group, 2.0 mg/ml group and the control group was 0 mg/ml). All the experiments were repeated three times. The cell morphologic and structure change was observed with the optical microscope and the transmission electron microscope. The proliferation of D341 cell was analyzed using Cell Counting Kit-8 assay. Apoptosis was detected by Annexin V-FITC/PI double staining. The expression of Caspase3 and Caspase9 was detected by Western blot.

RESULTSWith the effect of matrine, the proliferation inhibition rate gradually increased with drug concentrations increasing, and there was a significant difference (P < 0.01). The inhibitory effect of matrine on cell proliferation was different with the different treatment time, there was a significant difference between the 24 h to 72 h groups (P < 0.01). The apoptotic rate increased with matrine concentrations increasing. There were significant differences between the group of 0.5 mg/mI or 1.0 mg/mI to the group of 1.5 mg/mI or 2.0 mg/mI (P < 0.05). The apoptotic rate increased with the prolonged treatment time. There were significant differences between the group of 24 h or 48 h to the group of 72 h ( P < 0.05). With the increase of matrine concentration, the expression of Caspase 3 and Caspase 9 increased (P < 0.01).

CONCLUSIONMatrine induces the apoptosis, and inhibits the proliferation of human medulloblastoma D341 cells in vitro by up-regulation of the expression level of Caspase3, Caspase9.

Alkaloids ; pharmacology ; Apoptosis ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Cerebellar Neoplasms ; metabolism ; pathology ; Humans ; Medulloblastoma ; metabolism ; pathology ; Quinolizines ; pharmacology ; Up-Regulation

Objective To assess the value of multi-slice spiral CT angiography (MSCTA) in diagnosis of posterior nutcracker syndrome (NCS). Methods MSCTA of 15 patients clinically diagnosed as posterior NCS (patient group) and 50 subjects with normal renal vessels (control group) were retrospectively analyzed. The anatomy, course and relationship to the adjacent structure of left renal vein (LRV) and its branches were observed. The anteroposterior diameter (d1) and cross areas (s1) of the angle of control group and posterior abdominal artery (AA), the anteroposterior diameter (d2) and cross areas (s2) at the largest lumen near the renal hilar were measured and the ratio of s2/s1 and d2/ d1 were calculated.Results All LRVs posterior AA of patient group were compressed and narrowing with dilating apparently near the renal hilar. The difference of d2 was not significant (P＞0.05), but of other values were significant (P＜0.05).Conclusion MSCTA can show anatomy and three-dimentional trend clearly of LRV for posterior NCS, and evaluate the extent of narrow and dilating lumen accurately with cross areas measuring.

160 s and core temperature was 36-37℃.Blood ga- ses,platelets,plasma free hemoglobin and coagulation factors were measured during ECMO support. The blood supply was monitored in the limbs cannulated with the femoral artery cannula.Results Two patients were successfully treated with ECMO without major complications.The circulatory and respiratory function of the patients was stable.The chest X-ray showed a no clouding of lungs and he- patic function was greatly improved in case 2 who underwent a successful heart transplantation follo- wing mechanical cardiocirculatory support as a bridge,Two patients received ECMO support for 5 and 3 days respectively.Both patients recovered well.No any severe acute rejection occurred and heart func- tion was NYHAⅠ.Conclusion When candidates suffered acute cardiogenic shock,ECMO can provide safe and effective mechanical circulatory support as a bridge to heart transplantation.

Objective To investigate the effect of different Chinese herbs on cell proliferation and cartilage oligomeric matrix protein(COMP)expression in chondrocyte culture.Methods Chondrocytes isolated from rabbit knee cartilage were cultured for 3 generations with the density of 2?10~4/cm~2 and were verified by collagenⅡimmunohistochemical staining.Rabbit sera containing herbs were obtained after animals orally ad- ministrated herbs at the dosage equivalent to human.At 5% and 10% serum density,cells were cultured in the medium that contained liver-softening herbal compound sera.Subgroups setting at 1,3 and 5 hours after herb intervention were observed.Rabbit and bovine sera were control groups.Seven days after intervention,chon- drocytes proliferation was observed using the MTT assay kit.For the study of COMP expression,chondrocytes were isolated from human knee cartilage supematant.Superuatant COMP level was tested by enzyme-linked immunoabsorbent assays(ELISA)after directly adding compound and extract from liver-softening herbs to the culture at the final concentration of 10 mg/ml for 3 days.Results Liver-softening herbal compound group had significant effect on cell proliferation compared to control,of which,3-hour subgroup was more significant than 1-and 5-hour subgroups(P

In order to investigate the mechanism, the correlation between the odor change in Crataegi Fructus stir-fried process and 5-HMF were studied. Required samples were retrieved from Crataegi Fructus stir-fried process. Statistical quality control (SQC) was used to analyze the response values acquired by the electronic nose. At the same time, the content of 5-HMF was detected by high performance liquid chromatography (HPLC). Correlation analysis was used to analyze the relationship between the above two. Experimental results showed that SQC model established by response values of all samples could show the change law of odor in Crataegi Fructus stir-fried process and changes of 5-HMF content was dropped after the first increase. Correlation analysis showed that the odor change in Crataegi Fructus stir-fried process and 5-HMF were significantly correlated (P < 0.05). Sugar degradation reaction and the Maillard reaction may be one of the mechanisms of the odor change in Crataegi Fructus stir-fried process.
Chromatography, High Pressure Liquid ; Crataegus ; chemistry ; Furaldehyde ; analogs & derivatives ; analysis ; Hot Temperature ; Odorants ; analysis ; Plant Extracts ; analysis ; Technology, Pharmaceutical ; methods

Objective:To investigate the immune response after injecting polysaccharide nucleic acid of Bacillus Calmette-Guerin(BCG-PSN)under the mucous membrane of bladder in rabbits and to search for the most suitable dose of BCG-PSN.Methods:The rabbits were randomly divided into 5 groups:high dose(0.35 mg/ml)BCG-PSN group,mid- dle dose(0.175 mg/ml)BCG-PSN group,low dose(0.0875 mg/ml)BCG-PSN group,BCG(0.35 mg/ml)control group and BCG-PSN(0.35 mg/ml)intra-bladder perfusion control group.T lymphocyte subsets(CD4~+,CD8~+)in the peripheral blood were determined by flow cytometry before and 1 week,2 weeks,1 month and 3 months after the treatment; the levels of IL-2,TNF-?,and IFN-?of peripheral blood were determined by enzyme-linked immunosorbent assay;H-E and Masson staining was used for the pathological examination of the bladder.Results:(1)BCG-PSN injection increased the number of CD4~+ and CD8~+ T lymphocytes in a time- and dose-dependent manner,with the peak numbers appeared 2 weeks after BCG-PSN injection;the numbers restored to the normal levels 3 months after BCG-PSN injection.BCG control group had a similar changing pattern to the BCG-PSN injection group.The number of CD4~+ T cells BCG-PSN perfusion group was significantly lower than that in the high-dose BCG-PSN injection group(P

Objective To explore the effect of transplantation of mesenchymal stem cells (MSCs) transfected with the human receptor activity modifying protein 1 (hRAMP1) gene on proliferation and apoptosis of vascular smooth muscle cells (VSMCs) after carotid balloon angioplasty was performed in rabbits with carotid atherosclerosis.Methods Density gradient centrifugation and adherent culture were carried out to obtain MSCs,which were then transinfected with an adenovirus vector carrying the hRAMP1 gene or an empty adenovirus vector.A rabbit model of atherosclerotic stenosis and balloon angioplasty was successfully established.Results were randomly divided into three groups:the hRAMP1-MSCs group,theadipose-derived MSCs (Ad-MSCs) group and the control group.MSCs were transinfected with Ad-EGFP-hRAMP1,Ad-EGFP or PBS by transplantation into the injured carotid arteries.Homing and differentiation were assessed with MSCs harvested at 7 d.With MSCs collected at 28 d,Western blotting was used to measure the expression of the hRAMP1 target gene in the carotid artery; the neointima and media area in the injured carotid arteries were estimated; carotid artery morphology was examined with H&E staining; and the proliferation and apoptosis of VSMCs were determined by immunohistochemistry and TUNEL.Results The expression of CD31 and EGFP was found in proliferating neointima lesions at 7d in the hRAMP1-MSCs group and the Ad-MSCs group.At 28d of MSC transplantation,the level of RAMP1 significantly increased in the hRAMP1-MSCs group,compared with the Ad-MSCs and control groups [(63.0±4.9) vs.(28.3±2.5) and (27.2±7.2),all P＜0.05],but there was no differencein the RAMP1 level between the Ad MSCs group and the control group (P＞0.05).Positive expression of the α-smooth muscle antibody (α-SMA) was found in all three groups at 28 d of MSC transplantation.The thickness of the hyperplastic neointima significantly decreased in the hRAMP1-MSCs group,compared with the other two groups (P＜0.05),and was lower in the Ad-MSCs group than in the control group (P＜0.05).The expression of proliferating cell nuclear antigen (PCNA) was lower in the hRAMP1-MSCs group than in the Ad-MSCs and control groups at 28d of MSC transplantation (P ＜0.05),while the PCNA level was lower in the Ad-MSCs group than in the control group (P＜ 0.05).The VSMC apoptosis rate significantly increased in the hRAMP1-MSCs group,compared with the Ad MSCs and control groups (P＜0.05),and was the lowest in the control group (P＜0.05).Conclusions Gene-modified stem cell therapy can effectively inhibit vascular intimal hyperplasia,thereby reducing restenosis after angioplasty.

Objective To screen and analyze nucleotide variants in 5'-untranslated region(5'-UTR)in voltage-gated sodium channel α1-subunit gene(SCN1A)in patients with Dravet syndrome and to evaluate the association of the variants with disease.Methods Peripheral blood of 24 patients with Dravet syndrome and 100 unrelated normal persons were collected and genomic DNA was extracted.PCR-sequencing of SCN1 A 5'-UTR in these DNA was performed.To evaluate the possibility of mutation inducing disease,bioinformatics analysis was applied to analyze the conservation of the sequences around the mutation site and predict the potential transcription elements.Results The nucleotide variant of 166.642.520G→A in exon 2 was identified in two patients,but not in normal controls.The mutation was a de novo mutation in a patient with early-onset.In the second proband,the mutation was also carried by his clinically asymptomatic mother.The nucleotide site 166.642.520 was moderately conserved in mammals(62.5%).The average nucleotide identity rate between human and other mammals species in the region adjacent to 166.642.520 was 88.5%.Two potential transcription regulatory elements were predicted on the sequence with the mutation of 166.642.520G>A,and only one on the sequence with wild-type.Conclusions The mutation 166.642.520G>A may be associated with Dravet syndrome and further studied should be performed to verify it and demonstrate its pathogenic mechanisms.

Objective To investigate the relationship between uncoupling protein 2 (UCP-2) gene promoter -866 G/A polymorphism and Chinese diabetic nephropathy (DN). Methods A total of 182 patients with type 2 diabetic mellitus were selected and divided into DN group (90 cases with DN ) and NCD group (92 cases without DN ). Genomic DNA was detected, and UCP-2 genotype and allele gene frequency was confirmed by polymerase chain reaction-restrictive fragment length polymorphism, then compared.Results The genotype frequency of UCP-2 gene promoter-866 G/A polymorphism was distributed in DN group[GG 14.44%( 13/90),GA 31.11%(28/90),AA 54.44%(49/90)],and distributed in NCD group[GG 29.35% ( 27/92 ), GA 32.61% ( 30/92 ), AA 38.04% ( 35/92 )], and there was significant difference between two groups ( χ2 = 7.28 , P ＜ 0.05 ). There was also significant difference in allele gene frequency between DN group and NCD group[G 45.65% (84/184) vs. 30.00% (54/180),A 54.35% (100/184) vs. 70.00% (126/180)]( χ2 = 9.47, P ＜ 0.05 ). Conclusion There is correlation between the UCP-2 gene promoter-866 G/A polymorphism and Chinese DN, and the incidence of DN with A/A genotype is increased.

Objective To explore the clinical efficacy of the gap above the splenic pedicle in laparoscopic splenectomy (LS).Methods The retrospective cohort study was conducted.The clinical data of 189 patients who underwent LS in the Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology between August 2012 and March 2015 were collected.Among 189 patients receiving splenic pedicle division,42 without the application of the gap above the splenic pedicle were allocated into the group A and 147 with the application of the gap above the splenic pedicle were allocated into the group B.Observation indicators included:(1) operation situations:combined operation,operation time (excluding combined operation time),volume of intraoperative blood loss (excluding blood volume of spleen),cases with conversion to open surgery;(2) postoperative situations:time to initial anal exsufflation,time for fluid diet intake,occurrence of postoperative complications and duration of hospital stay;(3) follow-up.Patients were followed up by telephone interview and outpatient examination up to August 2016.Follow-up included routine blood test,coagulation function,liver function,with or without long-term complications.Measurement data with normal distribution were represented as x±s and comparison between groups was analyzed using the t test.Comparisons of count data were analyzed by the chi-square test.Results (1) Operation situations:of 189 patients undergoing LS,136 combined with laparoscopic pericardial devascularization,13 combined with laparoscopic cholecys-tectomy,9 combined with laparoscopic radio frequency ablation (RFA) of liver tumors and 26 combined with pathological examination using laparoscopic liver tissues sampling.Operation time,volume of intraoperative blood loss and cases with conversion to open surgery were (118±31) minutes,(80±38) mL,2 in the group A and (70± 22) minutes,(50± 28) mL,1 in the group B,respectively,with statistically significant differences between the 2 groups (t =12.579,-8.516,x2=4.912,P＜0.05).(2) Postoperative situations:time to initial anal exsufflation,time for fluid diet intake,number of patients with postoperative complications and duration of hospital stay were (22± 10)hours,(3.1 ± 1.3) days,8,(9±3)days in the group A and (23±11)hours,(3.8±1.8)days,13,(8±3)days in the group B,respectively.Pancreatic fistula,intra-abdominal hemorrhage,asymptomatic portal vein thrombosis,pulmonary infection and intraperitoneal infection were respectively detected in 2,2,2,1,1 patients in the group A and 1,2,5,2,3 patients in the group B.There was no significant difference in time to initial anal exsufflation,time for fluid diet intake and duration of hospital stay between the 2 groups (t =1.102,0.745,0.583,P＞0.05),and a statistically significant difference in number of patients with postoperative complications between the 2 groups (x2 =7.259,P＜ 0.05).There were statistically significant differences in cases with pancreatic fistula and intra abdominal hemorrhage (x2=16.021,5.812,P＜0.05) and no significant difference in cases with asymptomatic portal vein thrombosis,pulmonary infection and intraperitoneal infection (x2 =1.391,0.396,0.865,P＞0.05).Patients with postoperative complications were cured by symptomatic treatment.(3) Follow-up:156 of 189 patients (33 in the group A and 123 in the group B) were followed up for 1-18 months,with an average time of 12 months.During the follow-up,13 patients had recurrent hematemesis and melena,including 3 in the group A and 10 in the group B.Eight patients stopped bleeding after conservative treatment,3 stopped bleeding after proxial gastrectomy and 2 died of excessive bleeding and organ failure.Conclusion Splenic pedicle division using Endo-GIA through the gap above the splenic pedicle in LS can reduce operation time,volume of intraoperative blood loss,rate of conversion to open surgery and postoperative complications.