Roles of TSA upregulation miR-4298 targeting inhibition of PADI4 expression in inducing U251 cells apoptosis / 国际肿瘤学杂志

Objective:To investigate the mechanism of microRNA-4298 (miR-4298) targeting inhibition of peptidylarginine deiminase 4 (PADI4) expression in U251 cells apoptosis induced by histone deacetylase inhibitor trichostatin A (TSA).Methods:The experiment was divided into TSA group (0.2 μmol/L TSA) and contral group. CCK8 method was used to detect the effect of TSA on the proliferation of U251 cells. Flow cytometry was used to detect the apoptosis level of U251 cells after drug action. Reverse transcription PCR and Western blotting experiments were used to determine the changes of PADI4 gene and protein expression after miR-4298 interference. Luciferase assay was used to determine the effects of miR-4298 on targeted binding and luciferase activity in the 3′UTR region of PADI4. U251 cells were transfected with PADI4 to observe the rescue effect of miR-4298 on apoptosis. Each experiment was divided into 3 groups, NC group, miR-4298 mimic group, TSA+ miR-4298 mimic group and NC group, miR-4298 inhibitor group, TSA+ miR-4298 inhibitor group.Results:U251 cells were treated with 0.2 μmol/L TSA for 4 days, the Absorbancy ( A) values of the control group was 1.168±0.148, which was higher than those of the TSA group (0.737±0.007), with statistically significant difference ( t=4.948, P=0.008). Compared with the control group, after treatment with 0.2 μmol/L TSA for 48 h, U251 cells showed obvious apoptosis (27.62%±3.49% vs. 4.99%±0.13%, t=11.190, P<0.001). Compared with those before the drug treatment, the PADI4 gene expression (0.386±0.020 vs. 0.903±0.021) and protein expression (0.276±0.041 vs. 0.777±0.031) after TSA treatment for 48 h both were decreased, with statistically significant differences ( t=30.400, P<0.001; t=16.770, P<0.001). The expression of miR-4298 during the process of TSA-induced U251 cell apoptosis increased (2.573±0.289 vs. 1.003±0.136; t=8.487, P=0.001). There was a negative correlation between the miR-4298 expression and PADI4 expression ( r=-0.877, P=0.002). Luciferase experiment confirmed that miR-4298 has targeted binding and luciferase inhibitory effects on the 3′UTR region of wild-type PADI4. Compared with NC group (0.920±0.026), the relative expression levels of PADI4 gene of miR-4298 mimic group (0.413±0.049) and TSA+ miR-4298 mimic group (0.213±0.035) were decreased, with statistically significant differences (all P<0.001). The PADI4 protein expression level change was consistent with the gene change trend. Compared with NC group (3.78%±0.68%), the apoptosis levels of miR-4298 mimic group (7.96%±1.10%) and TSA+ miR-4298 mimic group (13.74%±1.26%) were increased, with statistically significant differences ( P=0.005; P<0.001). Compared with NC group (0.183±0.025), the PADI4 gene expression of miR-4298 inhibitor group (0.483±0.032) and TSA+ miR-4298 inhibitor group (0.386±0.025) were increased, with statistically significant differences ( P<0.001; P=0.015). The PADI4 protein expression level change was consistent with the gene change trend. Compared with NC group (4.96%±0.59%), the apoptosis levels of miR-4298 inhibitor group (23.83%±2.20%) and TSA+ miR-4298 inhibitor group (9.55%±1.49%) were increased, with statistically significant differences (all P<0.001). In the rescue experiment, the expression level of PADI4 in miR-4298 mimic group was significantly increased ( P<0.001), while the expression level of PADI4 in miR-4298 mimic+ PADI4 group was relatively reversed ( P=0.002). Conclusion:miR-4298 can participate in the process of U251 cell apoptosis by targeting the expression of PADI4 gene, and miR-4298 may be the target of targeted intervention therapy for glioma.

Study of transabdominal acoustic windows for the midsagittal plane of the fetal brain / 中华医学超声杂志（电子版）

Objective This study intends to evaluate different acoustic windows that can be used to obtain the midsagittal plane of fetal brain during abdominal ultrasound scanning and the application of acoustic windows in practice.Methods The midsagittal planes were obtained respectively through anterior fontanel,posterior fontanel,sagittal suture and forehead seam of 12 aborted fetuses,and feasible acoustic windows were used as standard acoustic windows to obtain midsagittal planes of second trimester screening fetuses,which were divided into two groups according to the position of fetal brain,cephalic presentation group (367 cases) and non-cephalic presentation group (384 cases).The success rates were compared between groups.Results Of the second trimester fetuses,497 midsagittal planes of 751 cases were successfully obtained.The success rate of the cephalic presentation fetuses (32.7％,120/367) was statistically lower than that of the non-cephalic presentation fetuses (98.2％,377/384,P ＜ 0.01).Of the cephalic presentation fetuses,the combined acoustic windows acquisition success rate of the forehead,anterior fontanelle,sagittal suture and posterior fontanel acoustic window was 32.7％ (32.7％,120/367),significantly higher than the single acquisition success rate of the forehead,anterior fontanelle,sagittal suture and posterior fontanel acoustic window (17.2％,25.9％,1.6％ and 7.6％,all P ＜ 0.01).Of the non-cephalic presentation fetuses,the combined acoustic windows acquisition success rate of the forehead,sagittal suture and posterior fontanel acoustic window was 98.2％ (98.2％,377/384),significantly higher than the single acquisition success rate of the forehead,anterior fontanelle,sagittal suture and posterior fontanel acoustic window (31.8％,61.2％,64.8％ and 62.0％,all P ＜ 0.01).Conclusions The midsagittal plane of fetal brain can be obtained through forehead seam,anterior fontanel,posterior fontanel and sagittal suture acoustic windows in transabdominal screening.The combination of four acoustic windows can increase the success rate of obtaining midsagittal planes.

Effect of byproducts in lignocellulose hydrolysates on ethanol fermentation by Issatchenkia orientalis / 生物工程学报

Byproducts in lignocellulose hydrolysates, namely sodium formate (1 to 5 g/L), sodium acetic (2.5 to 8.0 g/L), furfural (0.2-2 g/L), 5-hydroxymethylfurfural (5-HMF, 1 to 1.0 g/L) or vanillin (0.5 to 2 g/L) were used to evaluate their effects on ethanol fermentation by Issatchenkia orientalis HN-1 using single factor test and the response surface central composite experiment. Results showed that most of the byproducts had no obvious inhibition on the production of ethanol, except for the addition of 2 g/L vanillin or 1 g/L of 5-HMF, which reduced the ethanol production by 20.38% and 11.2%, respectively. However, high concentration of some byproducts in lignocellulose hydrolysates, such as sodium formate (1 to 5 g/L), sodium acetic (2.5 to 8.0 g/L), furfural (0.2 to 2 g/L) and vanillin (0.5 to 2 g/L) inhibited the growth of I. orientalis HN-1 significantly. Compared with the control, the dry cell weight of I. orientalis HN-1 decreased by 25.04% to 37.02%, 28.83% to 43.82%, 20.06% to 37.60% and 26.39% to 52.64%, respectively, when the above components were added into the fermentation broth and the fermentation lasted for 36 h. No significant interaction effect of the various inhibitors (sodium formate, sodium acetic, furfural and vanillin) except for vanillin single factor on the ethanol production was observed based on the central composite experiments. The concentrations of byproducts in most lignocellulose hydrolysates were below the initial inhibition concentration on ethanol production by Issatchenkia orientalis HN-1, which indicated that Issatchenkia orientalis HN-1 can be used for ethanol production from lignocellulose hydrolysates.

Feasibility of combining anterior and posterior fontanelle acoustic windows for median sagittal plane in fetal cranial ultrasound scanning / 中华围产医学杂志

Objective To explore the feasibility of combining anterior and posterior fontanelle acoustic windows for fetal median sagittal plane cranial scanning in prenatal ultrasound.Methods From July 5 to August 25 in 2012,200 pregnant women in Huai'an First Hospital at 22 to 24 gestational weeks accepted transabdominal uhrasonography,with anterior fontanelle,longitudinal suture and posterior fontanelle as acoustic windows to obtain the median sagittal plane.In the mean time,ten aborted or induced fetuses (with malformations or maternal complications) underwent cranial ultrasound with the same acoustic windows.The successful rates of obtaining median sagittal plane were calculated.Data were analyzed by Chi-square test.Results (1) Among the ten aborted or induced fetuses,all median sagittal planes were successfully obtained through anterior,posterior fontanelle,or longitudinal suture alone.(2) Among the 200 cases of prenatal uhrasonography,the successful rate of obtaining median sagittal plane through posterior fontanelle alone was 76.0％(152/200),which was higher than that through anterior fontanelle alone [59.5％ (119/200),x2=12.5,P＜0.01].The successful rate was 95.5％ (191/200) when combining the anterior and posterior fontanelle,which was higher than that through anterior fontanelle or posterior fontanelle alone (x2 =74.3 and 31.1,both P＜0.01).The successful rate of obtaining median sagittal plane through posterior fontanelle was 67.3％ (65/102) in cephalic presentation,which was also higher than that through anterior fontanelle [37.3％ (38/102),x2 =14.3,P＜0.01].The successful rate was 88.8％ (87/98) through posterior fontanelle in non-cephalic presentation,which was similar to that through anterior fontanelle [82.7％(81/98),x2 =1.5,P＞0.05].When both anterior and posterior fontanelle applied,the successful rate in cephalic presentation was 9.8％ (10/102),lower than that in non-cephalic presentation fetuses [71.4％ (70/98),x2 =79.1,P＜0.01].Conclusions The successful rate in obtaining median sagittal plane through posterior fontanelle is higher than that through anterior fontanelle,and might be increased when both anterior and posterior fontanelle were used.

Butanol production from corn stover hydrolysate with in-situ liquid-liquid extraction / 生物工程学报

Butanol production from corn stover hydrolysates (CSH) with in-situ liquid-liquid extraction was studied to enhance the production and reduce the fermentation cost. Oleyl alcohol was selected as the suitable solvent and added at the initial fermentation time with the ratio of 1:1 (oleyl alcohol: fermentation broth, V/V). Under this condition, butanol and ABE from CSH with 32.1 g/L total sugars were 3.28 and 4.72 g/L, which were 958.1% and 742.9% higher than those of the controls, respectively. Butanol and ABE production from CSH of 49.7 g/L total sugars after detoxification by ion exchange resin D301 coupled with extraction fermentation were 10.34 g/L and 14.72 g/L with an ABE yield of 0.31 g/g (g ABE/g utilized sugar), which were equal to those of glucose and xylose mixture fermentation. The detoxification and extraction fermentation technology of cellulosic butanol production would provide a crucial technical support to the industrialized production of cellulosic butanol.

Comparison of microorganisms fermenting syngas into ethanol / 生物工程学报

To evaluate the ability of microbial mix-culture fermenting syngas into ethanol, we studied the microbial mix-cultures A-fm 4, G-fm 4, Lp-fm 4 and B-fm 4 obtained by enrichment and compared with Clostridium autoethanogenum DSM10061 with 10% and 25% inoculation size. The results show that, with 10% inoculation size, the ethanol production of A-fm 4, G-fm 4, Lp-fm 4, B-fm 4 and C. autoethanogenum were 349.15, 232.16, 104.25, 79.90 and 26.99 mg/L respectively. With 25% inoculation size, the ethanol production were 485.81, 472.73, 348.58, 272.52 and 242.15 mg/L respectively. Higher inoculation size will increase the production of ethanol. The tested mix-culture exhibited a significant yield advantage compared with the maximum production of C. autoethanogenum reported in the literature (259.64 mg/L). This research provided a practical method to improve ethanol production from syngas.

Optimization of corn stover hydrolysis by fed-batch process / 生物工程学报

High-concentration sugars production from stover is an important perspective technology for the cellulosic ethanol industrialization. Fed-batch process is an effective way to achieve this goal in the fermentation industry. In this study, based on fed-batch process, high-concentration sugars were produced from pretreated corn stover by enzymatic hydrolysis. After being pretreated by the dilute sulphuric acid, the impacts of the ratio of solid raw material to liquid culture, the content of supplementary materials and the refilling time on the saccharification rate were investigated. Results showed that the initial ratio of solid raw material to liquid culture was 20% (W/V) and the initial concentrations of enzymes for xylanase, cellulose and pectinase were 220 U, 6 FPU, and 50 U per gram of substrates, respectively. After 24 hours and 48 hours, 8% pretreated corn stovers were added respectively together with the additions of xylanase (20 U) and cellulose (2 FPU) per gram of substrates. After 72 hours, the final concentration of reducing sugar was increased to 138.5 g/L from 48.5 g/L of the non fed-batch process. The rate of enzyme hydrolysis of the raw material was 62.5% of the thoretical value in the fed-batch process. This study demonstrated that the fed-batch process could significantly improve the concentration of reducing sugar.

Screening of high lipid production Trichosporon fermentans mutants by transposon tagging mTn-lacZ/leu2 insertion / 生物工程学报

To improve microbial lipid production, we inserted mTn-lacZ/leu2 into Trichosporon fermentans 2.1368-Leu(-) to obtain high lipid production mutants. By observing the LacZ chromogenic change, the positive reaction between Cerulenin (inhibitor of fatty acid synthase) and phosphate vanillin, a higher lipid-producing mutant 2.1368-Leu(-)-7 grown on corn-stalk hydrolysate was obtained. The lipid content of this mutant reached 38.30% (8.97% higher than that of the control) and the lipid production rate was 8.35% (20.63% higher than that of the control). The rate of sugar utilization was 77%, meaning that 100 g corn-stalk could be converted to 8.32 g lipid. The study provided an effective method for microbial lipid production by using cheap raw materials for biodiesel.

Effect of Electro-acupuncture on Expression of Transforming Growth Factor-β1 in the Cartilage of Experimental Rats with Knee Osteoarthritis / 中国康复理论与实践

@#Objective To investigate the effect of electro-acupuncture on cartilage renovation of knee osteoarthritis.Methods 40 female SD rats were randomly divided into the normal group,model group,electro-acupuncture treatment group and control group.The left knees of the treatment group and right ones of the control group were treated by electro-acupuncture for two weeks from the fifth week since operation.Collected left knees' cartilage of all rats at the seventh week since the beginning of experiment.HE and immunohistochemical staining were used to observe the expression characteristic of transforming growth factor-β1(TGF-β1)in cartilage.Results The expression of TGF-β1 in the treatment group was very significantly stronger than all other groups(P<0.01).Conclusion Electro-acupuncture can obviously up-regulate the level of TGF-β1 in cartilage of experimental rats with knee osteoarthritis,and promote the repair of cartilage.

Effect of Acupuncture on Different Traditional Chinese Medicine Syndrome Types in Patients with Apoplectic Hemiplegia / 中国康复理论与实践

@#Objective To observe effects of acupuncture on different traditional Chinese medicine(TCM) syndrome types in patients with apoplectic hemiplegia.Methods 57 cases of apoplectic hemiplegia were divided into the Qi deficiency with blood stagnation group(35 cases) and blockage of the meridians and collaterals by phlegm and blood group(22 cases).All patients were treated by scalp acupuncture combined with body acupuncture.The clinical therapeutic effect was evaluated according to the scores of manner,speech,motor function of limbs,and so on.Results In the Qi deficiency and blood stagnation group,2 cases were basically cured,17 were remarkably effective,12 were effective,with a total effective rate of 88.6%;while,in the blockage of the meridians and collaterals by phlegm and blood group,3 cases were basically cured,14 were remarkably effective,3 were effective,with a total effective rate of 90.9%.There was a significant difference between the two groups in clinical remarkably effective rate(P<0.01),but no difference in total effective rate( P>0.05).Conclusion Acupuncture can obviously improve the consciousness,language and limb function of patients with apoplectic hemiplegia.The effective rate of type of blockage of the meridians and collaterals by phlegm and blood is superior to type of Qi deficiency with blood stagnation.

Effect of Scalp Acupuncture Combined with Traditional Body Acupuncture on Pseudobulbar Palsy Induced by Stroke / 中国康复理论与实践

@#ObjectiveTo observe clinical therapeutic effect of scalp acupuncture combined with traditional body acupuncture on pseudobulbar palsy induced by ischemic or hemorrhagic apoplexy.Methods59 inpatients with pseudobulbar palsy induced by stroke were divided into the scalp acupuncture combined with body acupuncture group (n=40) and body acupuncture group (n=19). Cases of the scalp acupuncture combined with body acupuncture group were treated by scalp acupuncture combined with traditional body acupuncture. Cases of the body acupuncture group were treated only by traditional body acupuncture. Therapeutic effect of two groups was assessed according to patients' consciousness, language and limb functions.ResultsIn the scalp acupuncture combined with body acupuncture group, 30 cases were cured and 9 were remarkably effective, with a total effective rate of 97.5%. While in the body acupuncture group, 9 cases were cured, and 7 were remarkably effective, with a total effective rate of 84.2%. There was a significant difference between two groups in clinical cure rate (P<0.05), but no difference in total effective rate ( P>0.05).ConclusionScalp acupuncture combined with traditional body acupuncture can obviously improve the consciousness, language and limb functions and pseudobulbar palsy of stroke patients and obtain a better effect compared with traditional body acupuncture.

脊髓损伤患者焦虑和抑郁倾向调查及护理对策 / 中国康复理论与实践

@#目的分析脊髓损伤患者的心理特点,指导临床心理护理。方法采用焦虑自评量表(SAS)及抑郁自评量表(SDS)对86例脊髓损伤住院患者进行评定,根据得分高低分别归纳出焦虑和抑郁的主要表现,并制定有针对性的心理护理计划。结果与结论74.4%的患者有焦虑情绪,77.9%有抑郁情绪。焦虑的主要表现为夜间睡眠不好、常常要小便、手脚常常湿冷、不容易安静坐着、觉得会发生什么不幸等,抑郁的主要表现为性功能障碍、做事情感到困难、不容易下决定、生活无意义、吃饭比平时减少等。脊髓损伤患者普遍存在抑郁及焦虑情绪 ,应有针对性地进行心理康复护理。