Optimization of the conditions for the synthetic preparation of iron sucrose complex by response surface methodology / 国际药学研究杂志

Objective To optimize the conditions for the synthetic process of iron sucrose complex (ISC), via the investigation of the effects of reaction temperature (X1), reaction time (X2), amount of alkali (X3), and amount of sucrose (X4) on the relative molecular mass of the ISC product. Methods According to the experimental results for the single factor, the conditions dealing with the X1, X2, X3, and X4 parameters for the preparation of ISC were optimized by the Box-Behnker design combined with the response surface methodology using the weight average relative molecular mass of ISC as an indicator, and analyzed with gel permeation chromatography. Results The reaction temperature and the amount of alkali had a significant effect on the weight average relative molecular mass of ISC. The influence of the four factors in the descending order was as follows:X3＞X1＞X2＞X4. In the designed experimental conditions, theresponsevaluedecreasedwiththeincreaseofbothreactiontemperaturesandalkaliamounts. Conclusion Theresponse surface methodology could provide the relationship between the response values and variables via the minimum number experiments to obtain the optimized conditions for the preparation of ISCs.

Risk Factors for Mesh Exposure after Transvaginal Mesh Surgery / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Mesh exposure after surgery continues to be a clinical challenge for urogynecological surgeons. The purpose of this study was to explore the risk factors for polypropylene (PP) mesh exposure after transvaginal mesh (TVM) surgery.</p><p><b>METHODS</b>This study included 195 patients with advanced pelvic organ prolapse (POP), who underwent TVM from January 2004 to December 2012 at the First Affiliated Hospital of Chinese PLA General Hospital. Clinical data were evaluated including patient's demography, TVM type, concomitant procedures, operation time, blood loss, postoperative morbidity, and mesh exposure. Mesh exposure was identified through postoperative vaginal examination. Statistical analysis was performed to identify risk factors for mesh exposure.</p><p><b>RESULTS</b>Two-hundred and nine transvaginal PP meshes were placed, including 194 in the anterior wall and 15 in the posterior wall. Concomitant tension-free vaginal tape was performed in 61 cases. The mean follow-up time was 35.1 ± 23.6 months. PP mesh exposure was identified in 32 cases (16.4%), with 31 in the anterior wall and 1 in the posterior wall. Significant difference was found in operating time and concomitant procedures between exposed and nonexposed groups (F = 7.443, P = 0.007; F = 4.307, P = 0.039, respectively). Binary logistic regression revealed that the number of concomitant procedures and operation time were risk factors for mesh exposure (P = 0.001, P = 0.043).</p><p><b>CONCLUSION</b>Concomitant procedures and increased operating time increase the risk for postoperative mesh exposure in patients undergoing TVM surgery for POP.</p>

Relationship of cystatin C, fibrinogen, and 24-hour urinary protein with renal pathological grade in children with Henoch-Schönlein purpura nephritis / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the relationship of cystatin C (CysC), fibrinogen (Fbg), and 24-hour urinary protein with renal pathological grade in children with Henoch-Schönlein purpura nephritis (HSPN), and to explore their values.</p><p><b>METHODS</b>The clinical data of 48 children diagnosed with HSPN by renal biopsy from January 2011 to January 2015 were reviewed. According to renal pathological grading, in the 48 children with HSPN, 12 had stage IIa or lower, 12 stage IIb, 17 stage IIIa, and 7 stage IIIb or higher. The latex-enhanced immunoturbidimetric assay, turbidimetric measurement, and end-point method were used to determine the levels of serum CysC, Fbg, and 24-hour urinary protein, respectively. Pearson and Spearman correlation analyses were used to test the correlations between the indices and between the indices and renal pathological grade.</p><p><b>RESULTS</b>There were significant differences in the levels of serum CysC, Fbg, and 24-hour urinary protein between patients with different pathological grades (P<0.05). The level of each index increased with increasing pathological grade (P<0.05). In the 48 children with HSPN, the level of 24-hour urine protein was positively correlated with the levels of serum CysC (r=0.51, P<0.05) and Fbg (r=0.63, P<0.05). The level of Fbg was positively correlated with that of serum CysC (r=0.55, P<0.05). The levels of CysC, Fbg, and 24-hour urinary protein were all positively correlated with renal pathological grade (r=0.66, 0.64 and 0.68; respectively, P<0.05).</p><p><b>CONCLUSIONS</b>The levels of CysC, Fbg, and 24-hour urine protein can reflect the severity of renal injury, providing satisfactory prediction of the severity of renal injury in children with HSPN.</p>

Prediction of microRNA-296-5p target genes and its application in lung development / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To predict the target genes of rno-microRNA-296-5p (miR-296) using bioinformatics software and databases, and to provide a theoretical basis for further studies of biological effects of miR-296 in fetal lung development.</p><p><b>METHODS</b>PubMed and Google were used to search for all reported literature on miR-296. The miRBase database was used to determine the sequence and evolutionary conservatism of miR-296. The TargetScans database was used to predict the target genes of miR-296. The DAVID Bioinformatics Resources 6.8 database was used for the functional enrichment analysis of the target genes. The KEGG database was used to analyze the signaling pathways of target genes.</p><p><b>RESULTS</b>miR-296 was reported to play important roles in many biological processes and have a high degree of sequence conservation among species. The target genes of miR-296 were involved in biological processes, cell components, and molecular function. Those target genes were significantly enriched in the mitogen-activated protein kinase signaling pathway, Wnt signaling pathway, and transforming growth factor-β signaling pathway (p<0.05).</p><p><b>CONCLUSIONS</b>The bioinformatics analysis of the target genes of miR-296 provides a basis for studying biological effects and mechanism of action of miR-296 in lung development.</p>

Immunohistochemical expression of HO-1 in traumatic human brain tissue / 法医学杂志

OBJECTIVE@#To investigate the expression of heme oxygenase-1 (HO-1) at different intervals and to provide evidence for estimation on injury intervals after brain contusion in human.@*METHODS@#Twenty-four patients died of serious brain injury were assigned as injury group and 4 patients died of non-brain injury were served as control group. HO-1 expression was analyzed in brain tissue at different time intervals (3 h, 6-9 h, 12-24 h, 36 h-3d, 5-8d, 17-20d) by immunohistochemistry and auto-image analysis system.@*RESULTS@#The level of HO-1 expression started to increase in 3 h after brain contusion compared to the control group (P < 0.05). The level of HO-1 expression highest level in 12-24 h group, and maintained high level in 36 h-3 d, then decreased gradually.@*CONCLUSION@#The expression of HO-1 might be a strong evidence for human brain contusion time estimation.

The simultaneous knock-down of Ku70 and Ku80 by a tandem Ku-shRNA-encoding plasmid expression system / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To extend the use of vector-derived siRNA by generating multiple shRNAs in the same plasmid.</p><p><b>METHODS</b>Construct a vector that expresses shRNAs targeting on Ku70 and Ku80 in tandem. The gene silencing efficiency of each shRNA was verified previously. After identification by restriction digestion and DNA sequencing, the reconstructed plasmid, named psiRNAKus, was transfected into the human hepatoma cell line HepG2. The tandem-shRNA-induced silencing of targeted genes was determined by RT-PCR at RNA level and Western blot at protein level.</p><p><b>RESULTS</b>The shRNAs encoded by psiRNAKus down-regulated both the expression of Ku70 and Ku80.</p><p><b>CONCLUSION</b>The vector-derived siRNA delivery system that allows multiple shRNA species to be expressed from the same vector may be of value in experimental and therapeutic applications.</p>

A method using long primers for cloning the upstream sequence of delta-6 fatty acid desaturases gene of Thamnidium elegans by nested inverse PCR / 生物工程学报

Thamnidium elegans is a kind of phycomycete that produces essential unsaturated fatty acids, particularly y-linolenic acid. In this process, delta6-Fatty acid desaturase (D6D) plays a key role due to its enzymatic properties that catalyze the delta6 site dehydrogenation of precursor linoleic acid (18:2delta(9, 12) n-6) and a-linolenic acid (18:3delta(9, 12, 15) n-3). This reaction is the first and rate-limiting step of highly unsaturated fatty acids (HUFA) synthesis pathways. After we have isolated and cloned the gene coding delta6-fatty acid desaturase from Thamnidium elegans As3.2806 (GenBank accession number DQ099380), our interest focuses on the promotion and regulation of the gene transcription. To achieve this aim, we designed long primers and used nested inverse PCR to amplify DNA flanking sequences. First, genome of Thamnidium elegans was extracted and digested with restriction enzymes EcoR I and Kpn I , respectively. Then we ligated the digested DNA with T4 ligase at low concentration which is propitious for linear DNA to joint intromolecule. According to the sequence of delta6-fatty acid desaturase gene of Thamnidium elegans, we designed a couple of 35nt long inverse primers and two couples of shorter inverse primers for inverse PCR. Three rounds of PCR reactions were performed. In the primary reaction, the ligated DNA was used as a template, and the product was used as the template of the secondary reaction, the tertiary reaction was achieved in the same way. After all the three rounds of reactions, we got a nice product about 4 kb from the EcoR I digested sample, in which a 1.3kb 5' upstream sequence (GenBank accession number DQ309425) of delta6-fatty acid desaturase gene containing several putative regulatory elements including TATA. box, FSE-2, AP-1 sites, CCAAT cis-element site and STRE-binding site was derived after sequencing. All of these implied intensely that this 1.3kb fragment is a condition-regulated promoter. It is the first report about Thamnidium elegans detla6-fatty acid desaturase gene promoter. The procedure described here is a rapid and simple method and particularly useful to isolate flanking sequences from fungal genome. box, FSE-2, AP-1 sites, CCAAT cis-element site and STRE-binding site was derived after sequencing. All of these implied intensely that this 1.3 kb fragment is a condition-regulated promoter. It is the first report about Thamnidium elegans delta6-fatty acid desaturase gene promoter. The procedure described here is a rapid and simple method and particularly useful to isolate flanking sequences from fungal genome.

High vaginal uterosacral ligament suspension for treatment of uterine prolapse / 中华妇产科杂志

Objective To evaluate the physiological and anatomic basis,indications,surgical skills, prevention of ureter injury and clinic outcomes of using high uterosacral ligament suspension(HUS)for correction of advanced uterine prolapse by the vaginal route.Methods Fifty women with advanced uterine prolapse underwent transvaginal HUS after vaginal hysterectomy with reconstruction of pubocervical and rectovaginal fascia to correct their uterine prolapse between June 2003 and September 2007.The average age of the women was 60.1 years.The mean follow-up period was 24 months(range 4-51 months).The degree of pelvic organ prolapse preoperatively and anatomic outcomes postoperatively were assessed with pelvic organ prolapse quantification system(POP-Q).Results The remnants of the uterosacral ligaments were clearly identified and palpated posterior and medial to the ischial spines by traction with a 24 cm long Allis clamp and used for successful vaginal vault suspension and reconstruction in all 50 consecutive advanced uterine prolapse patients.The ureter injury was avoided by complete knowledge of the ureter's course from the cervix/apex toward its insertion in the sacral region and how far outside of the uterosacral ligament,by uteri palpation and by suturing purposefully placed"deep"dorsally and posteriorly toward the sacrum,as well as by cystoscopy examination of the spillage of urine from both ureters.Mean POP-Q point C improved from 1.5 to-7.5 cm with a median follow-up of 24 months.If the successful HUS was defined as point C≤stage I prolapse,both the objective and subjective cure rates were as high as 100% with a maximum follow-up of 51 months.None of the 50 patients had repeat operation for recurrence of prolapse.There was no major intra-or postoperative complications,such as ureter and other pelvic organ injury.Conclusion HUS with fascial reconstruction seems to be a safe,minimal traumatic,tolerable and highly successful procedure for vaginal repair of advanced uterine prolapse.Because of the use of native tissue as suspension site HUS is more physiologic and cost effective.

A Modified Method for Isolating DNA from Fungus / 微生物学通报

Genomic DNA of two fungi Thamnidium elegans and Umbelopsis isabellina were extracted with an amended Cetyltrimethyl Ammonium Bromide (CTAB) method. This modified method uses repeated freezing in liquid nitrogen and thawing with combination of shocking with glass beads to replace of the tra- ditional method. Quality and concentration of DNA extracted by the modified methodwere tested. Compared with the traditional method, higher yield and purity of genomic DNA were obtained with less amount ofmy- celium. The result indicted that this is a simple and highly efficient method, which is suitable to treat many samples at one time and for basic molecular experiments, such as restriction endonuclease reaction and PCR.