Objective To investigate expressions of microRNA-21 (miR-21),PTEN,NF-κB,and caspase-9 in colon carcinoma and their possible mechanisms in progression of tumor.Methods Expressions of above-mentioned indexes and percentage of apoptotic cells in colon carcinoma,colon adenoma,and normal tissue specimen were detected.Results Expressions of miR-21,PTEN,N F-κB,and caspase9 in normal tissue were 0.39 ±0.02,50％,25％,and 50％.Expressions of them in colon adenoma were 0.62 ±0.06,50％,35％,and 55％,and those in colon carcinoma were 0.88 ±0.04,30％,75％,and 20％.Percentage of apoptotic cells were (24.64 ± 7.66) ％,(15.86 ± 1.44) ％,and (6.20 ± 2.57) ％,respectively.The above P-values were all less than 0.05.Expression of miR-21 was positively correlated with TNM stage,depth of infiltration,differentiated level and expression of NF-κB while inversely correlated with PTEN and caspase-9 expressions and percentage of apoptotic cells.The r-values were 0.647,0.297,0.259,0.519,-0.299,-0.388,and-0.931,respectively.Conclusions Hyperexpression of miR-21 participated in progression of colon carcinoma probably through down-regulating expressions of PTEN and caspase-9 and up-regulating expression of NF-κB and then suppressing cell apoptosis.

Objective To investigate the application opportunity and effect of B-Lynch suture in severe postpartum hemorrhage during cesarean section. Methods One hundred patients with high risk to postpartum hemorrhage were randomly divided into two groups. Group A was given with preventive treatment by B-Lynch suture after the placenta was born. Group B was given with traditional treatment, such as kneading the uterus, pressing, "8" suture, stuffing with gauze first. B-Lynch suture was used after the failure of traditional treatment or the amount of bleeding was over 400 ml.At the same time, another 50 patients without severe postpartum hemorrhage were selected as control (group C). Observed the difference of the amount of bleeding during different time, the operation time and comphcations among the three groups. Results The amount of bleeding during operation and 2 h after childbirth in group A [(266.00±29.90),(301.33±40.30) ml] were obviously less than those in group B[(512.67±202.12), (554.17±201.78)ml](P<0.01). The operation time in group A [(33.73±2.28)min] was shorter than that in group B [(49.33±10.40) min] (P<0.01). The incidence rate of severe postpartum hemorrhage in group A (6%, 3/50) was lower than that in group B (64%, 32/50) (P<0.01). Every parameter in group A was similar with group C. There were no serious complications during and after operation. Conclusions The preventive use of B-Lynch suture in patients with high risk to postpartum hemorrhage attains much better effects, and depresses the incidence rate of severe postpartum hemorrhage, and also improves the living quality. B-Lynch suture should be. used widely in patients with high risk to postpartum hemorrhage preventively.

Objective To determine the amount of serum Vitamin D in premature infants,and to investigate its correlation with bone quantitative ultrasound measurement.Methods The serum of premature infants born between 2013 March and 2014 March in the Maternal and Children Health Hospital of Huadu District in Guangzhou were collected,and serum 25-hydroxyvitamin D [25-(OH) D] level was measured by using chemiluminescence immunoassay,while Omnisense quantitative ultrasound was used to measure bone speed of sound(SOS) in the middle area of the left tibia.According to gestational age,the participants were divided into A,B and C groups(28-32 weeks,32 +1-34 weeks,34 + 1-36 +6 weeks,respectively).The levels of 25-(OH)D and SOS were compared and the correlation between them was analyzed.Results The amount of 25-(OH) D of the 3 groups was (41.65 ± 21.15) nmoL/L,(47.15 ± 19.78) nmol/L,and (49.35 ± 19.93) nmol/L,respectively,and the differences among the 3 groups were statistically significant (F =4.441,P =0.012).The ratio of vitamin D abundant or not (insufficiency including deficiency and lack) in preterm among the 3 groups were compared,and the differences among the 3 groups were statistically significant(x2 =11.38,P =0.023).SOS of the 3 groups were (2 787.85 ± 123.01) m/s,(2 865.12 ± 129.44) m/s and (2 908.59 ± 124.01) m/s,respectively,and the differences among the 3 groups were statistically significant (F =28.716,P =0.000).There was a positive correlation between 25-(OH) D and SOS (r =0.084,P =0.024).Conclusions Level of Vitamin D in premature infants is generally inadequate.The smaller the gestational age,the higher the occurrence rate.Vitamin D levels and SOS are significantly positively correlated,and both of them increase with gestational age.

Objective To investigate the methods and application effectiveness of micro-lecture in the clinical nursing specialty courses.Methods The micro-video was empoldered with camera tool to record,record the screen software production,PPT directly method,etc.The micro-video was applied in teaching practice of clinical nursing specialty courses.Seven class students in 2013 in Nursing of Guangxi Medical University were selected as teaching object randomly and were divided into the experimental group and the control group with 49 cases cach.The experimental group was used micro-lecture aided teaching,the control group was used traditional teaching.After the class,the teaching effect was evaluated by the examination,clinical probation appraisal and evaluation questionnaire of the experimental group students.The data were statistically analyzed by SPSS 11.5.Results The results of theory examination in the experimental group was (87.20 ± 6.50) scores and in the control group was (75.30 ± 7.89) scores,there was significant difference,t=3.20,P ＜ 0.01.The qualified result of clinical probation appraisal in the experimental group was 93.88％ (46/49),and in the control group was 81.63％(40/49),there was significant difference,x2=6.712,P＜ 0.01.The teaching satisfaction of micro-lecture in the experimental group was better.Conclusion The microlecture which based on the micro video as the main carrier used in teaching can effectively improve the effect of teaching,and is a valuable teaching method as a supplementary of the traditional teaching.

Objective To study the correlation between the expressions of Bcl-2 and Caspase-3 in intestinal epithe-lial and inflammatory cells of lamina propria and the staging and pathogenesis of ulcerative colitis (UC). Methods The pathomorphological changes of 25 samples of UC in the active phase and remission phase and 25 samples of control were ob-served under microscope. The expressions of Bcl-2 and Caspase-3 were detected by immunohistochemistry SP method. The correlations between expressions of Bcl-2, Caspase-3 and UC staging were analyzed. Results There were no significant dif-ferences in expressions of Bcl-2 and Caspase-3 in intestinal epithelial cells between three groups (P>0.05). The expression of Bcl-2 in inflammatory cells of lamina propria was significantly higher in active UC group than that of normal control group (P＜0.05), no significant difference between UC remission group and normal control group (P>0.05). There was a higher positive expression of Caspase-3 in inflammatory cells of lamina propria in UC remission group. There was correlation be-tween Bcl-2, Caspase-3 and UC staging (r=0.371 and 0.4453 respectively). Conclusion The expression levels of Bcl-2 and Caspase-3 were related with pathogenesis of UC. The detection of expression levels of Bcl-2 and Caspase-3 has a cer-tain application value in UC staging.

Objective To study the correlation between TCM differentiation of syndromes typing for ulcerative colitis (UC) and expressions of Bax and Caspase-3.Methods 50 patients with UC were divided into 2 groups according to TCM syndrome differentiation including dampness-heat internal accumulation syndrome group (25cases),deficiency of spleen and stomach syndrome group (25cases).Immunohistochemistry was used to study expression of Bax and Caspase-3 in colonic mucosas and compare with the normal group (25cases).Results The expressions of Bax were highest in dampness-heat internal accumulation syndrome group (4.56±1.58).The expressions of Bax of deficiency of spleen and stomach syndrome group were lower than dampness-heat internal accumulation syndrome group(3.28± 1.14)scores.There were significant differences between the three groups (P＜0.05).However,There were no significant difference between dampness-heat internal accumulation syndrome group and deficiency of spleen and stomach syndrome group in terms of the Caspase-3expressions (P＞0.05).In addition,Caspase-3 and Bax were positively related in each group(r=0.23and 0.21).Conclusion The high expressions of Bax,Caspase-3 in ulcerative colitis were closely related to TCM typing according to syndrome differentiation.Caspase-3 and Bax were adjustment in apoptosis.The detection of expression of Bax,Caspase-3 is helpful to the TCM syndrome differentiation and to the selection of treatment plan.At the same time,it can provide more important experiment basses for the pathogenesis of ulcerative colitis.

9α-hydroxy-4-androstene-3,17-dione (9-OH-AD) is an important intermediate in the steroidal drugs production. 3-ketosteroid-9α-hydroxylase (KSH), a two protein system of KshA and KshB, is a key-enzyme in the microbial steroid ring B-opening pathway. KSH catalyzes the transformation of 4-androstene-3,17-dione (AD) into 9-OH-AD specifically. In the present study, the putative KshA and KshB genes were cloned from Mycobacterium smegmatis mc(2)155 and Gordonia neofelifaecis NRRL B-59395 respectively, and were inserted into the expression vector pNIT, the co-expression plasmids of kshA-kshB were obtained and electroporated into Mycobacterium sp. NRRL B-3805 cells. The recombinants were used to transform steroids, the main product was characterized as 9α-hydroxy-4-androstene-3,17-dione (9-OH-AD), showing that kshA and kshB were expressed successfully. Different from the original strain Mycobacterium sp. NRRL B-3805 that accumulates 4-androstene-3,17-dione, the recombinants accumulates 9α-hydroxy-4-androstene-3,17-dione as the main product. This results indicates that the putative genes kshA, kshB encode active KshA and KshB, respectively. The process of biotransformation was investigated and the results show that phytosterol is the most suitable substrate for biotransformation, kshA and kshB from M. smegmatis mc(2)155 seemed to exhibit high activity, because the resultant recombinant of them catalyzed the biotransformation of phytosterol to 9-OH-AD in a percent conversion of 90%, which was much higher than that of G. neofelifaecis NRRL B-59395. This study on the manipulation of the ksh genes in Mycobacterium sp. NRRL B-3805 provides a new pathway for producing steroid medicines.
Androstenedione ; analogs & derivatives ; biosynthesis ; Bacterial Proteins ; genetics ; metabolism ; Biotransformation ; Ketosteroids ; Mixed Function Oxygenases ; genetics ; metabolism ; Mycobacterium ; metabolism ; Mycobacterium smegmatis ; enzymology ; Plasmids

Objective: Study on the improvement of quality standard for Zicao ointment. Methods: The content of shikonin and β,β’-dimethylacrylshikonin was determined by high performance liquid chromatogram (HPLC). The column was Inertsil ODS-3 C18 column (4.6 mm×250 mm, 5 μm). The mobile phase comprising of acetonitrile-0.05% formic acid solution (70:30). The flow rate was 1.0 ml/min, the detection wavelength was 275 nm, and the column temperature was 25 ℃. Results: The linear range of shikonin and β,β’-dimethylacrylshikonin were 0.04-0.81 μg (r=0.999 5) and 0.41-10.36 μg (r=0.999 2). The average recovery were 103.32% and 101.80%, the RSD were 1.96% and 2.11%. Conclusions The method is simple, accurate and sensitive to control the quality of Zicao ointment.

Objective To investigate the expressions of microRNA-145 (miR-145) and Six1 in gastric cancer tissues and gastric cancer cell lines,and to investigate the effect of miR-145 targeting Six1 on invasion of human gastric cancer cells and its molecular mechanism.Methods Sixty patients with advanced gastric cancer confirmed by gastroscopy and pathology in the Fourth Hospital of Changsha were selected,from January to November,2017.The expressions of miR-145 mRNA and Six1 in 60 cases of gastric cancer and their matched adjacent tissues were detected by real-time fluorescence quantitative PCR (qRT-PCR) and their correlations were analyzed.miR-145 mimics were transfected into MKN-45,BGC-823 and SGC-7901 gastric cancer cell lines.The relative levels of miR-145 mRNA in three gastric cancer cell lines were detected by qRT-PCR.The SGC-7901 gastric cancer cell lines with the highest expression level of miR-145 mRNA were selected for subsequent experiments.The mimic group (transfected with miR-145 mimic),NC group (transfected with miR-145 negative control) and empty carrier group (no addition of any oligonucleotides) were set up.The invasive ability of gastric cancer cells was detected by Transwell test.The ability of angiogenesis was verified by microtubule formation experiment.The expression levels of downstream proteins of the gastric cancer cell lines SGC-7901 such as Six1,E-cadherin and vimentin were detected by Western blotting method.The MirTrap system and the luciferase report test verified whether miR-145 targeted the Six1 gene.Results The expressions of miR-145 mRNA in gastric cancer tissues and in para cancerous tissues were 0.579 ± 0.086 and 1.009 ±0.121,respectively.The difference was statistically significant (t =-22.498,P ＜0.001).The expressions of Six1 were 2.516 ± 0.208 and 1.041 ± 0.227,respectively.The difference was statistically significant (t =37.119,P ＜ 0.001).The expressions of miR-145 mRNA and Six1 were negatively correlated (r =-0.728,P ＜0.001).After overexpression of miR-145,in the mimic,NC and empty carrier groups,the numbers of cells passing through the membrane were 48.550 ± 3.716,82.800 ± 3.797,87.467 ± 8.023,respectively.The difference was statistically significant (F =87.789,P ＜ 0.001).There were significant differences between the mimic and NC groups and between the mimic and empty carrier groups (both P ＜0.001).In the mimic,NC and empty carrier groups,the numbers of tube formation of human umbilical vein endothelial cells were 24.333 ± 1.211,34.167 ±2.041,36.500 ±3.209,respectively.The difference was statistically significant (F =47.103,P ＜ 0.001).There were significant differences between the mimic and NC groups and between the mimic and empty carrier groups (both P ＜ 0.001).In the mimic,NC and empty carrier groups,the expression levels of Six1 in gastric cancer SGC-7901 cells were 1.392 ±0.072,2.426 ±0.099,2.371 ±0.079,respectively.The difference was statistically significant (F =289.517,P ＜ 0.001).There were significant differences between the mimic and NC groups and between the mimic and empty carrier groups (both P ＜0.001).In the mimic,NC and empty carrier groups,the expression levels of E-cadherin were 4.001 ±0.132,2.714 ±0.181,2.653 ±0.218,respectively.The difference was statistically significant (F =106.572,P ＜0.001).There were significant differences between the mimic and NC groups and between the mimic and empty carrier groups (both P ＜ 0.001).In the mimic,NC and empty carrier groups,the expression levels of vimentin were 1.634 ± 0.132,3.349 ± 0.102,3.501 ± 0.185,respectively.The difference was statistically significant (F =389.032,P ＜0.001).There were significant differences between the mimic and NC groups and between the mimic and empty carrier groups (both P ＜ 0.001).MirTrap system verified that the target Six1 mRNA levels in the mimic,NC and empty carrier groups were 1.101 ± 0.097,0.582 ± 0.037,0.573 ± 0.032,respectively.The difference was statistically significant (F =138.922,P ＜ 0.001).There were significant differences between the mimic and NC groups and between the mimic and empty carrier groups (both P ＜0.001).Luciferase reporter assay showed that the double fluorescence ratios of Six1 wild type recombinant vector group were 7.324 ± 0.415,10.755 ± 0.481,10.430 ± 0.309 in the mimic,NC and empty carrier groups respectively.The difference was statistically significant (F =129.345,P ＜ 0.001).There were significant differences between the mimic and NC groups and between the mimic and empty carrier groups (both P ＜0.001).The double fluorescence ratios of Six1 mutant recombinant vector group were 10.938 ± 0.091,11.077 ±0.126,11.028 ±0.205 in the mimic,NC and empty carrier groups respectively,and the difference was not statistically significant (F =1.318,P =0.297).The MirTrap system and the luciferase report test showed that miR-145 was targeted to the Six1 gene.Conclusion The expressions of miR-145 mRNA and Six1 in gastric cancer and para cancerous tissues were different and negatively correlated.miR-145 inhibited the invasion,angiogenesis and epithelial mesenchymal transition of gastric cancer cells by targeting Six1 gene.