Simultaneous Determination of 7 Effective Components in Yao Medicine Yueli Yaomi Spray by QAMS / 中国药房

OBJECTIVE：To establish QAMS method for simultaneou s determination of 7 effective components in Yao medicine Yueli yaomi spray ，such as α-cyperone，α-pinene，β-pinene，limonene，β-elemene，caryophyllene oxide and ligustilide ， so as to provide method reference for the quality control of the preparation. METHODS ：GC method was adopted. The determination was performed on DB- 1701P capillary column ，using nitrogen as carrier gas. The temperature of the hydrogen flame ion detector was 240 ℃. The temperature was programmed ，the inlet temperature was 240 ℃，the injection volume was 1 μL and the split ratio was 20 ∶ 1. Using limonene as internal reference ，the relative correction factors of other 6 components were calculated，the contents of them were calculated with relative correction factors ，and then compared with the results of internal standard method （using naphthalene as internal standard ）. RESULTS ：The mass concentration linear range of α-cyperone， α-pinene，β-pinene，limonene，β-elemene，caryophyllene oxide and ligustilide were 0.008 9-1.110 0，0.028 3-3.540 0，0.020 5- 2.560 0，0.023 0-2.880 0，0.016 3-2.035 0，0.013 1-1.640 0，0.008 3-1.040 0 mg/mL（all r＞0.999 0）；the limits of quantification were 0.005 6，0.013 1，0.011 4，0.018 6，0.010 8，0.008 9，0.004 5 mg/mL；the detection limits were 0.001 9，0.004 1，0.003 7， 0.006 2，0.003 5，0.002 9，0.001 5 mg/mL；RSDs for precision ，stability（24 h），and repeatability tests were all less than 2％ （n＝5 or n＝6）； the average recoveries were 98.48％ ， 014） 101.37％，97.96％，99.80％，102.79％，97.77％，102.14％， and RSDs were all lower than 2％（n＝9），respectively. The average relative correction factors of α-cyperone，α-pinene， β-pinene，β-elemene，caryophyllene oxide and ligustilide were 1.045 8，0.621 0，0.488 5，0.382 9，0.708 9，0.956 9 respectively，and the RSDs were all lower than 2％（n＝6）. There wa s no statistical significance in contents of 7 components between QAMS method and internal standard method （P＞0.05）. CONCLUSIONS ：The established QAMS method is simple ， accurate，stable and reproducible ，and can be used for simultaneous determination for 7 components in Yueli yaomi spray.

An overview of tools for post-analysis of high-throughput sequencing data in clinical study / 中华医学遗传学杂志

With the advance of high-throughout sequencing technology and its extensive application in clinical diagnosis, analysis of sequencing data has become an important part of clinical diagnosis. To date, the development and establishment of various software and databases have made it convenient to extract useful information from massive amounts of high-throughput sequencing data. However, it is still a challenge for correlating the clinical-genetic diagnosis based on the above-mentioned sequence data with the screened DNA variations and disease phenotypes. Further validation of the proposed pathogenesis with the discovered molecular defects are required. Here a comprehensive review is provided for the strategies of sequencing data analysis, commonly used phenotype-genotype correlation tools, and functional analysis and verification methods for the genetic diagnosis.

Advances in research on gene and cell therapy for type IV collagen related hereditary kidney diseases / 中华医学遗传学杂志

Type IV collagen is a component of the extracellular matrix in the basement membrane. Abnormal secretion or assembly of type IV collagen may lead to kidney lesions resulting in numerous nephropathy symptoms, e.g., Alport syndrome, thin basement membrane nephropathy, and focal segmental glomerulosclerosis. Treatment for type IV collagen-related nephropathy includes drugs, kidney transplantation, gene and cell therapy. However, drugs are not always effective, and kidney transplantation is hindered by the shortage of donors. Moreover, basement membrane nephritis often occurs after kidney transplantation. Therefore, gene and cell therapy probably is the most promising treatment for type IV collagen related nephropathies.

Changes of meibomian gland function and the stability of tear film after long-term wearing orthokeratolog / 中华实验眼科杂志

Objective This study was to observe the meibomian gland scores,the miss rate of meibomian,break-up time of tear film (BUT) changes after long-term wearing orthokeratolog,and to evaluate orthokeratolog impact on meibomian gland function and the stability of the tear film.Methods A prospective case-controlled study was performed.Fifty patients 98 eyes with myopia were enrolled.According to the myopia correction mode,the patients were divided into control group (24 cases) and experimental group (overnight orthokeratolog treatment) group (26 cases).All subjects were underwent the examinations sequentially as follows:evaluation of ocular surface disease symptoms using the Ocular Surface Disease Index (OSDI),lid margin and ocular surface examination by slit lamp microscrope,infrared meibomian photography and BUT.All measurements were repeated at 3 months,6 months,24-months follow-up visit.All patients signed informed consent before examination.Results After wearing orthok-contact lens for 24 months,the meibomian gland score was 4.54±1.10 and 2.29±0.75 in the experimental group and control group,respectively,the difference was statistically significant (P＜0.05).The BUT was (6.35 ± 1.52) s 24 months after wearing orthokeratolog,which was less than (9.38 ± 1.88) s in the control group,the difference was statistically significant (P＜ 0.05).The miss rate of meibomian gland was 24.39 ± 5.54 and 22.81 ± 5.23 in the experimental group and control group,respectively.There was no statistically significant difference among the the miss rate of meibomian gland between groups and within groups (all at P＞0.05).Conclusions Long-term wearing orthokeratolog has negetive effects on meibomian gland function and the stability of the tear film.

Effect of electroacupuncture and herbal cake- partitioned moxibustion on anxiety and depression in patients with Crohn’s disease in remission / 针灸推拿医学（英文版）

Objective:To observe the effect of electroacupuncture (EA) and herbal cake-partitioned moxibustion on anxiety and depression in patients with Crohn’s disease (CD) in remission. Methods:Sixty CD cases were randomly allocated into an EA group (n=30) and an herbal cake-partitioned moxibustion group (n=30) using the random number table by the ratio of 1:1. In addition, 30 healthy subjects were included in a control group. Bilateral Tianshu (ST 25), Qihai (CV 6) and Zhongwan (CV 12) were used in the EA and herbal cake- partitioned moxibustion groups. The treatment was done 3 times a week, for a total of 12 weeks. The efficacy was evaluated using self-rating anxiety scale (SAS), self-rating depression scale (SDS) and traditional Chinese medicine (TCM) symptom scores. Results:Before treatment, the SAS and SDS scores in CD patients were remarkably higher than those in healthy subjects. After EA or herbal cake-partitioned moxibustion treatment, the SAS and SDS scores were significantly decreased in both groups, showing significant intra-group differences (P0.05). Conclusion:Both EA and herbal cake-partitioned moxibustion can significantly decrease abnormally high SAS and SDS scores in CD patients as well as TCM symptom scores. The two therapies share similar effects in alleviating common symptoms and improving anxiety and depression.

miR-486-5p enhances senescence of human mesenchymal stem cells by de-creasing telomerase activity / 中国病理生理杂志

AIM:To investigate the effects of microRNA-486-5p (miR-486-5p) on the senescence of human mesenchymal stem cells ( hMSCs).METHODS: The expression of miR-486-5p was determined by miRNA arrays and real-time PCR.By transfection of miR-486-5p mimic or inhibitor, up-regulation or down-regulation of miR-486-5p expres-sion in hMSCs was established.The effect of miR-486-5p and silence information regulator 1 (SIRT1) on hMSC telomerase activity and senescence were detected byβ-galactosidase staining.RESULTS:The expression of miR-486-5p was up-regu-lated in the old hMSCs compared with the young hMSCs.Up-regulation of miR-486-5p resulted in increasing senescence of hMSCs.Conversely, down-regulation of miR-486-5p resulted in decreasing cell senescence.The expression of SIRT1 and telomerase reverse transcriptase ( TERT) was down-regulated in the old hMSCs compared with the young hMSCs.Directly repression of SIRT1 expression inhibited the hMSC TERT protein expression and telomerase activity, but increased cell se-nescence.The regulation of miR-486-5p on hMSC senescence was attenuated by inhibiting the expression of miR-486-5p and SIRT1 together.CONCLUSION:miR-486-5p enhances senescence of hMSCs by decreasing the expression of SIRT1 and telomerase activity.

Progress in the tumorigenic potential of iPSCs and methods to reduce it / 生物工程学报

Since Yamanaka successfully reprogrammed murine fibroblasts into iPSCs in 2006, iPSCs technology has drawn much attention worldwide. Although iPSCs provides tremendous possibilities for both basic research and regenerative medicine, it has meanwhile potential risks, e.g. tumorigenicity. Scientists, therefore, have made efforts in clarifying the mechanism of the cause for iPSCs tumorigenicity and the way how to reduce the risk. The results of some researches reveal some of tumorigenic factors, e.g. the partial similarity of gene expression profiles between cancer cells and iPSCs, the accumulation of the genetic damages in the course of reprogramming process, and mutation in the cellular culture. As a consequence, numerous methods for reducing iPSCs tumorigenicity have been explored, such as minimized use of the reprogramming factors at the controlled manner, and the selection of the expression vector or parental cells. In this paper, the cause of iPSCs tumorigenicity and the current achievements on preventing iPSCs tumorigenesis are reviewed.

Analysis of drug resistance on extended spectrum β-lactamases-producing Escherichia coli and Klebsiella pneumoniae / 国际检验医学杂志

Objective Analysis of clinical distribution and susceptibility to antimicrobial agents in producing extended spectrumβ-lactamases (ESBLs)of Escherichia coli(ECO)and Klebsiella pneumoniae (KPN).Methods The samples from 2013 April to 2014 December in Changji Prefecture People′s Hospital were cultured and identified,drug sensitivity test did by VITEK2compat au-tomatic bacteria analysis,phenotypic confirmatory test was did on producing ESBLs ECO and KPN,and the clinical distribution and drug resistance was analyzed.Results 1 576 strains of ECO and KPN isolated 306 strains of producing ESBLs strains and 343 strains of multi drug resistant bacteria,the isolation rates were 1 9.4% and 21.8% respectively.In 576 strains of ECO,there were 1 77 strains of producing ESBLs strains and 202 strains of multi drug resistant bacteria were isolated,the separation rates were 30.7% and 35.1% respectively,generic drug resistant strains were not found,and those producing ESBLs strains were isolated mainly from the urine specimens (39.5%),and mainly from the ICU (37.3%)and (29.9%)the surgical ward.There were 129 strains of producing ESBLs strains,141 strains of multi drug resistant bacteria and 4 generic drug resistant strains were isolated from 1 000 strains of KPN,the separation rates were 12.9%,14.1% and 0.4% respectively,and those producing ESBLs strains were isolated mainly from sputum specimens (83.7%),and mainly from the ICU (42.6%)and(29.5%)in the department of inter-nal medicine.Isolation of producing ESBLs ECO and multi drug resistant bacteria were higher than KPN,the difference were statis-tically significant (P <0.05).The sensitivities of ECO and KPN ESBLs strains to carbapenems imipenem andaminoglycosides ami-kacin and compound preparation piperacillin/tazobactam were good,the other antimicrobial drug resistance rates were more than 50.0%.Conclusion ESBLs ECO mainly causes urinary tract infections,KPN mainly causes respiratory tract infection.Detection rates of producing ESBLs ECO and KPN were high,and the multi drug resistance is serious,should strengthen the monitoring of drug resistance on producing ESBLs strains,and according to the results of drug susceptibility,standardize the rational use of anti-microbial agents,and to strengthen the management of antimicrobial agents,delay the occurrence of bacterial resistance.

Analysis of distribution and drug resistance of Enterococcus faecium / 中国医师进修杂志

Objective To investigate the distribution and drug resistance ofEnterococcus faecium,provide evidence for clinical rational use of antimicrobial agents.Methods Retrospective analysis of drug resistance of Enterococcus faecium,summarize the specimen type,distribution of departments and so on,to explore the trend of drug resistance.Results Enterococcus faecium to vancomycin resistance rate from 3.54％ (4/113) in 2010,rose to 8.45％ (18/213) in 2013.One strains of hnezolid resistant Enterococcus faecium 717 was found,the minimum inhibitory concentration of 7.9 mg/L,not tigecycline resistance in Enterococcus faecium found.Conclusion Vancomycin resistant Enterococcus faecium for linezolid and tigecycline susceptibility is still high,the two kinds of antimicrobial agents can be used for the treatment of vancomycin resistant Enterococcus faecium.

Establishment of the methodology for quantifying lentiviral vector transcriptional read-through rate / 生物工程学报

As an effective vehicle for bio-research and for gene therapy, Lentiviral Vector (LV) has been drawn large attention in recent years. However, transcriptional read-through limits its application. In order to understand the extend of LV read-through in chromosome, a reliable method to assess transcriptional read-through rate is needed. Here, we report the method as follows: 293T cells were transfected with the lentiviral transfer vectors which borne with two LTRs at its two ends in order to mimic the state of "proviral vectors" in chromosome. Using the primers specific for 3'U5 and 3'U3, read-through and total transcripts were reverse transcribed, respectively. These two cDNAs were quantified by realtime PCR using the primers and probe specific for 5'end of 3'U3. Read-through rate was then calculated by the division of the two. Meanwhile, read-through product of green fluorescence protein was also analyzed by Fluorescence Activated Cell Sorter. They both reciprocally proved the principal and confirmed that self-inactivated LV appeared higher read-through rate than the wild type one. The method described in this article, therefore, provides a useful technique to study how to reduce read-through rate, and improve the bio-safety of LV.

Progress in improvement of lentiviral vectors' transcriptional read-through / 生物工程学报

Four out of 10 patients of X-linked severe combined immunodeficiency (X-SCID) were finally developed leukemia after receiving the treatment of gene therapy delivered by gamma-retroviral vectors. This is due to the vector integrated to the proximity of lmo2 etc proto-oncogene promoters, leading to the activation of onco-gene expression, which raises the concern of the bio-safety of gene therapy vectors. Lentiviral vectors, especially self-inactivating lentiviral vectors, are considered to be much safer than gamma-retroviral vectors. However self-inactivating lentiviral vectors also have encountered with some unsafe factors and one of them is the problem of transcriptional "read-through" . During the past years, achievements have been made to reduce lentiviral vector transcriptional read-through, which are reviewed herein.