A novel homozygous frameshift variant in DNAH8 causes multiple morphological abnormalities of the sperm flagella in a consanguineous Pakistani family / 亚洲男科学杂志(英文版)

Multiple morphological abnormalities of the sperm flagella (MMAF) is a severe form of asthenozoospermia categorized by immotile spermatozoa with abnormal flagella in ejaculate. Whole-exome sequencing (WES) is used to detect pathogenic variants in patients with MMAF. In this study, a novel homozygous frameshift variant (c.6158_6159insT) in dynein axonemal heavy chain 8 (DNAH8) from two infertile brothers with MMAF in a consanguineous Pakistani family was identified by WES. Reverse transcription-polymerase chain reaction (RT-PCR) confirmed DNAH8 mRNA decay in these patients with the DNAH8 mutation. Hematoxylin-eosin staining and transmission electron microscopy revealed highly divergent morphology and ultrastructure of sperm flagella in these patients. Furthermore, an immunofluorescence assay showed the absence of DNAH8 and a reduction in its associated protein DNAH17 in the patients' spermatozoa. Collectively, our study expands the phenotypic spectrum of patients with DNAH8-related MMAF worldwide.

A recurrent homozygous missense mutation in CCDC103 causes asthenoteratozoospermia due to disorganized dynein arms / 亚洲男科学杂志(英文版)

Asthenoteratozoospermia is one of the most severe types of qualitative sperm defects. Most cases are due to mutations in genes encoding the components of sperm flagella, which have an ultrastructure similar to that of motile cilia. Coiled-coil domain containing 103 (CCDC103) is an outer dynein arm assembly factor, and pathogenic variants of CCDC103 cause primary ciliary dyskinesia (PCD). However, whether CCDC103 pathogenic variants cause severe asthenoteratozoospermia has yet to be determined. Whole-exome sequencing (WES) was performed for two individuals with nonsyndromic asthenoteratozoospermia in a consanguineous family. A homozygous CCDC103 variant segregating recessively with an infertility phenotype was identified (ENST00000035776.2, c.461A>C, p.His154Pro). CCDC103 p.His154Pro was previously reported as a high prevalence mutation causing PCD, though the reproductive phenotype of these PCD individuals is unknown. Transmission electron microscopy (TEM) of affected individuals' spermatozoa showed that the mid-piece was severely damaged with disorganized dynein arms, similar to the abnormal ultrastructure of respiratory ciliary of PCD individuals with the same mutation. Thus, our findings expand the phenotype spectrum of CCDC103 p.His154Pro as a novel pathogenic gene for nonsyndromic asthenospermia.

Novel biallelic loss-of-function mutations in / 亚洲男科学杂志(英文版)

Multiple morphological abnormalities of the sperm flagella (MMAF) is a specific type of asthenoteratozoospermia, presenting with multiple morphological anomalies in spermatozoa, such as absent, bent, coiled, short, or irregular caliber flagella. Previous genetic studies revealed pathogenic mutations in genes encoding cilia and flagella-associated proteins (CFAPs; e.g., CFAP43, CFAP44, CFAP65, CFAP69, CFAP70, and CFAP251) responsible for the MMAF phenotype in infertile men from different ethnic groups. However, none of them have been identified in infertile Pakistani males with MMAF. In the current study, two Pakistani families with MMAF patients were recruited. Whole-exome sequencing (WES) of patients and their parents was performed. WES analysis reflected novel biallelic loss-of-function mutations in CFAP43 in both families (Family 1: ENST00000357060.3, p.Arg300Lysfs*22 and p.Thr526Serfs*43 in a compound heterozygous state; Family 2: ENST00000357060.3, p.Thr526Serfs*43 in a homozygous state). Sanger sequencing further confirmed that these mutations were segregated recessively in the families with the MMAF phenotype. Semiquantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) was carried out to detect the effect of the mutation on mRNA of the affected gene. Previous research demonstrated that biallelic loss-of-function mutations in CFAP43 accounted for the majority of all CFAP43-mutant MMAF patients. To the best of our knowledge, this is the first study to report CFAP43 biallelic loss-of-function mutations in a Pakistani population with the MMAF phenotype. This study will help researchers and clinicians to understand the genetic etiology of MMAF better.

Effect of Electroporation on Percutaneous Permeation of Sinomenine Hydrochloride / 中国实验方剂学杂志

Objective:To investigate the percutaneous permeability of sinomenine hydrochloride （SNH） and optimize the parameters of electroporation to achieve the best permeation enhancing effect on SNH. Method:The percutaneous permeability of SNH and the enhancement effect of electroporation were studied by <italic>in vitro</italic> diffusion cell method， and the enhancement effect of electroporation was further evaluated by <italic>in vivo</italic> study in mice. Result:Under steady-state condition， the permeation rates of SNH in stripped skin and intact skin of hairless mice were （385.81±12.88）， （0.88±0.20） μg·cm^-2·h^-1， respectively. The permeation rate in stripped skin was 438 times higher than that in intact skin. The results of percutaneous permeation kinetics analysis showed that the solubility and diffusion coefficient of SNH in stratum corneum were relatively low， which were （70.82±9.63）×10³ g·m^-3 and （3.07±1.52）×10^-14 cm²·s^-1， respectively. Under the optimized electroporation conditions （voltage of 72 V， time of 60 min）， the 24 h cumulative permeation amount of SNH through skin of mice was （10 008.39±1 961.57） μg·cm^-2， and the steady-state permeation rate was （456.01±51.26） μg·cm^-2·h^-1， which were 5.4 times and 5.1 times higher than those of blank group， respectively. <italic>In vivo</italic> studies in mice showed that the contents of SNH in skin and muscle of electroporation group were 2.0 times and 1.5 times higher than those of blank group. Conclusion:The low solubility and low diffusion coefficient of SNH in the stratum corneum are the main factors hindering the percutaneous permeation of SNH. Electroporation can significantly increase the percutaneous permeation of SNH and its retention in skin and muscle of mice.

Research progress on biological activities of Olea europaea leaf extract / 中国中药杂志

Olea europaea oil is one of the most important part of the "Mediterranean dietary pattern", and a lot of epidemiological evidences showed that people with the Mediterranean diet having a lower morbidity of the cardiovascular system diseases, skin cancer and colon cancer. The health benefits of a Mediterranean diet not only attributed to monounsaturated fatty acids and vitamins and other nutrients in O. europaea oil, but also the phenolic compounds named as antioxidant effect. Studies have shown that O. europaea leaf contains much more antioxidant activity composition than the fruit, and oleuropein, flavonoids such polyphenols are the main active ingredients in O. europaea leaf. A small amount of O. europaea was introduced into China in 1956, after widely cultivated in Fujian, Guangdong, Taiwan, Sichuan, Shaanxi, Yunnan, and Longnan in Gansu province is the biggest O. europaea planting area in the country. In every winter pruning O. europaea will produce a large number of the leaves, which could be a high added value products (phenolic compounds) of rich source. This article through consulting the literature at home and abroad, classified and summarized the biological activity research status of O. europaea leaf extract and the possible mechanisms, including antimicrobial, antitumor, antioxidation, and on the function of brain, cardiovascular system, anti-diabetes, anti-inflammatory and analgesia and so on. At the same time looked ahead to its development prospects of O. europaea leaf extract, it has variety and high content of active ingredients, and antioxidant synergy, which provide a theoretical basis for the further development and utilization of O. europaea leaf. And O. europaea leaf extract has a rich cheap source and good bioavailability, which provided a broad space in the application of medical and health care.

Risk and Precautions against Single Dispensing in Emergency Pharmacy / 中国药房

OBJECTIVE:To reduce the risk of single dispensing,and to improve the quality of pharmaceutical care in emergen-cy pharmacy. METHODS:Single dispensing environmental factors,human factors and social factors of emergency pharmacy in our hospital were analyzed;the suggestions and measures were put forward,risk intervention mechanism was established and manage-ment effects were evaluated. RESULTS:Through improving the pharmacy hardware and software,carrying out the relevant sys-tem,enhancing training,improving the process of dispensing and window order,promoting the drug management and carrying out near error management. Compared with before management(Oct.-Dec. 2014),the times of near error and disputes significantly re-duced after management(Feb.-Apr. 2015)(P<0.05),and medication error reduced to zero. CONCLUSIONS:As single dispens-ing,risk management and the improvement of prescription audit ability can improve pharmaceutical care in emergency pharmacy.

Analysis of Gram-positive bacterial infection in patients following liver transplantation / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Liver transplantation is the most effective treatment for patients with end-stage liver failure, however infection after transplantation is a serious clinical complication. The purpose of this research was to investigate the molecular epidemiology and the influence of multidrug-resistant Gram-positive infection in patients, following liver transplantation, to provide reference for clinical treatment and prevention of Gram-positive bacterial infection.</p><p><b>METHODS</b>We isolated and detected bacteria from phlegm, throat swabs, urine, wound or wound secretions, blood, and fecal samples from 221 liver transplant patients in our hospital from January 2007 to April 2010. All isolated bacterial strains were identified and tested by minimal inhibitory concentration (MIC) drug-sensitive detection using the BioMerieux ATB bacterial identification instrument and repetitive extragenic palindromic-polymerase chain reaction (REP-PCR) detection of bacterial homology. Risk factors were calculated by multivariate Logistic regression analysis.</p><p><b>RESULTS</b>We collected 250 specimens from 221 patients hospitalized following liver transplantation surgery, of which 29 patients developed multiple infections. Sixty-five Gram-positive bacterial strains were isolated from different specimens from 53 infectious patients. We detected 29 multidrug-resistant Gram-positive strains from 29 patients (44.62%), including 20 Staphylococcus aureus (S. aureus) strains (68.97%) and nine Enterococcus strains (31.03%). All 20 S. aureus strains were highly resistant to aminoglycosides (gentamicin), cephalosporins (cefoxitin), quinolones (ciprofloxacin and levofloxacin), lincomycins (clindamycin), penicillin, and erythromycin. The resistance rate reached 100% in some cases. The S. aureus strains were highly sensitive to vancomycin and oxazolidinone (linezolid), with MIC50 < 2 µg/ml for both. The nine Enterococci strains were also highly resistant to aminoglycosides, quinolones, and penicillins, and highly sensitive to vancomycin (MIC50 < 2 µg/ml) and oxazolidinone (MIC50 < 1 µg/ml). Using REP-PCR detection, S. aureus was divided into five genotypes with 14 B-type strains. Enterococcus was divided into 11 genotypes, with two D-type strains, two G-type strains, and two K-type strains. The risk factors for Gram-positive bacterial infection in patients following liver transplantation were preoperative use of antibiotics (OR = 3.949, P = 0.004), high intra-operative blood input (OR = 1.071, P = 0.005), and postoperative renal failure (OR = 5.427, P = 0.043).</p><p><b>CONCLUSIONS</b>S. aureus and Enterococcus were the main pathogens causing infection following liver transplantation in patients with drug-resistant Gram-positive bacterial infection. The isolated strains were resistant to multiple antibiotics. B-type S. aureus strains were predominant. Reasonable use of antibiotics, decreasing intra-operative blood input, and preventing post-operative renal failure may reduce Gram-positive bacterial infections and the appearance of drug-resistant strains following liver transplantation.</p>

Oleanolic acid inhibits proliferation of HUVECs, and inhibits migration and tube formation via VEGF pathway / 药学学报

To investigate the effects of oleanolic acid (OA) on the proliferation, migration and the formation of tube-like structure in human vascular endothelial cells (HUVECs). MTT assay, flat plate scarification, Transwell plates and matrigel-induced tube formation assay were performed to detect the effects of OA on proliferation, migration and tube formation. MTT assay showed that the inhibition rates of HUVECs treated with 60 and 100 microg x mL(-1) of OA for 24 h were 19% and 83% respectively. Treatment of HUVECs significantly inhibited the cell migration in a dose-dependent manner. The vascular indexes of HUVECs treated with 40 and 60 microg x mL(-1) OA were 33% and 20% respectively. Western blotting analysis showed that treatment of the cells with OA significantly attenuated the expression and secretion of VEGF. Additionally, VEGF could in part reverse the effects of OA on migration and tube formation of HUVECs. In conclusion, OA inhibits the proliferation, and VEGF plays an important role in OA induced decreased migration and tube formation of HUVECs.

Effect of He-Ne laser verus low-level CO2 laser irradiation on accelerating fracture healing / 中国组织工程研究

BACKGROUND: The effects of low-level laser biostimulation have been proved by various experimental studies and clinical application, which are manifested as tissue repairing, analgesia, antiinflammation, etc.OBJECTIVE: To observe the effects of He-Ne laser and low-level CO2 laser irradiation on fracture healing.DESIGN: A randomized controlled animal experiment. SETTING: Research Room of Laser Medicine, Chengde Medical College.MATERIALS: Forty-eight healthy male New Zealand rabbits, weighing 2.0-2.5 kg, were used.METHODS: The experiments were carried out in the Research Room of Laser Medicine, Chengde Medical College from 1998 to 2003. ①The 48 rabbits were induced into models of experimental fracture of the left radius, and then they were divided into three groups with 16 rabbits in each group: control group, He-Ne laser irradiation group and CO2 laser irradiation group. The fracture areas of the animals in the He-Ne and CO2 laser irradiation groups were respectively irradiated with 28 mW/cm2 He-Ne laser and 150 mW/cm2 CO2 laser for 10 minutes, once a day. ② The animals were killed on the 15th and 35th days after taking roentgenogram respectively. At 15 days, the collagen and calcium contents in callus were detected. At 35 days, the biomechanic anti-torsion strength was examined.MAIN OUTCOME MEASURES: ①Results of roentgenogram in each group; ② Collagen and calcium contents in callus; ③ Results of biomechanic anti-torsion strength.RESULTS: Totally 45 rabbits were involved in the analysis of results. ① On the 15th and day after fracture, the roentgenogram, collagen and calcium contents in callus in the He-Ne and CO2 laser irradiation groups were better than those in the control group, and the collagen and calcium contents in callus were higher in the CO2 laser irradiation group than in the He-Ne laser irradiation group [(341.9±30.1) vs (302.1±28.7) mg/g; (197.1±19.7)vs (156.5±17.6) mg/g, P＜ 0.05]. ②On the 35th days after fracture, the biomechanic anti-torsion functions in the He-Ne and CO2 laser irradiation groups were superior to that in the control group, and there were no significant difference between the two laser irradiation groups. ③ The results of roentgenogram in the He-Ne and CO2 laser irradiation groups were better than that in the control group both on the 15th and 35th days after fracture, and there was no significant difference between the two irradiation groups.CONCLUSION: Both CO2 laser and He-Ne laser irradiations can accelerate the process of fracture healing, and the curative effect of CO2 laser is better than that of He-Ne laser on the 15th day after fracture, but there is no significant difference clinically between the CO2 laser and the He-Ne laser irradiations on the 35th day after fracture.

Protection of oxyphenamone on myocardium against ischemia-reperfusion injury in rat heart / 药学学报

<p><b>AIM</b>To study the protective effect of oxyphenamone, a novel inodilator against myocardial ischemia-reperfusion injury.</p><p><b>METHODS</b>A model of regional myocardial ischemia-reperfusion injury was established by ligating the left anterior desending coronary artery (LAD) in rat heart 10 min followed by reperfusion 15 min in vitro or 30 min in vivo. The protective effects of oxyphenamone were evaluated from the incidence of arrhythmia and the changes of myocardial creatine phosphokinase (CPK) and lactate dehydrogenase (LDH) activities, malondialdehyde (MDA) content, and myocardial ultrastructure.</p><p><b>RESULTS</b>In preparations of rat Langendorff hearts, infusion of oxyphenamone (1-10 micromol.L(-1)) diminished the incidence of ventricular fibrillation, decreased the activities of CPK and LDH in coronary efflux, and antagonized the increase of MDA content in ischemic myocardium significantly. The ischemia-reperfusion injury in anesthetized rats produced severe ventricular arrhythmia, decrease of CPK in myocardium, increase of CPK in serum, increase both of LDH and MDA both in myocardium and in serum, and severe damage of myocardial ultrastructure. Intravenous injection of oxyphenamone 0.1-1.0 mg.kg(-1) 5 min before ischemia ameliorated dose-dependently ventricular arrhythmia, antagonized the changes of CPK, LDH and MDA in both myocardium and serum induced by ischemia-reperfusion. It even maintained these parameters at normal level. The effects were somewhat similar to that of verapamil 1.0 mg.kg(-1) Intravenous injection of oxyphenamone 0.5 or 1.0 mg.kg(-1) 5 min after ligation of LAD also antagonized the ischemia-reperfusion induced changes in CPK, LDH and MDA in myocardium and serum significantly, and ameliorated the damage of myocardial ultrastructure markedly. The therapeutic effects of oxyphenamone were similar to that of propranolol 2. 0 mg.kg(-1).</p><p><b>CONCLUSION</b>From the examination of ECG, myocardial enzymes and ultrastructure, it appears that oxyphenamone can protect myocardium against ischemia-reperfusion injury induced by occlusion of LAD both in vitro and in vivo.</p>