Cdc42-dependent endocytosis pathway in the regulation of Na+/H+exchanger 3 (NHE3) expression on rotavirus-infected Caco-2 cells / 中华微生物学和免疫学杂志

Objective To observe the effects and regulatory mechanism of rotavirus infection on the expression and bioactivity of Na+/H+exchanger 3 (NHE3) on Caco-2 cells. Methods A cell model of Caco-2 cells expressing NHE3 was constructed. Four groups were set up,which were control(CTL) group, rotavirus(RV) infection group, Cdc42 inhibitor (Pirl-1) group and Pirl-1+RV group. Bioactivity and ex-pression of NHE3 on the surface of Caco-2 cells were determined by BCECF-AM and biotinylation method, respectively. Expression of Cdc42 protein was measured by Western blot. Co-immunoprecipitation was per-formed to detect the interaction between NHE3 and Cdc42. Results Compared with the CTL group,RV in-fection significantly inhibited the bioactivity and expression of NHE3 on Caco-2 cells. These inhibitory effects were antagonized by Pirl-1. Moreover,RV infection enhanced the expression of Cdc42 protein and promoted the interaction between NHE3 and Cdc42, which were also antagonized by Pirl-1. Conclusion RV infec-tion might regulate the expression and bioactivity of NHE3 through Cdc42-dependent endocytosis pathway.

Analysis of the Utilization of Oxycodone and Acetaminophen in Outpatient and Emergency Department of Our Hospital during 2013-2015 / 中国药房

OBJECTIVE:To provide reference for rational use of oxycodone and acetaminophen in outpatient and emergency department. METHODS:By retrospective study,the application of oxycodone and acetaminophen in outpatient and emergency de-partment of our hospital during 2013-2015 was analyzed statistically. RESULTS:During 2013-2015,there were 22654 outpatient and emergency prescriptions of oxycodone and acetaminophen in our hospital. The prescription number,amount and consumption sum of oxycodone and acetaminophen were increasing year by year. Top 3 clinical departments of oxycodone and acetaminophen in the list of prescription number were orthopedics department(78.34%),emergency department(15.52%)and oncology department (2.09%). The age of patients using oxycodone and acetaminophen mainly aged 19-64 years old(78.62%);the ratio of male to fe-male was 1:1.08. The amount of oxycodone and acetaminophen was 5 mg,mainly 4 times a day(86.91%). In all kinds of diagno-sis,knee osseous arthritis (47.26%),pain in waist and lower extremities (6.18%) and abdominal pain in dispute (5.92%) were top three. CONCLUSIONS:A large amount of oxycodone and acetaminophen were used in outpatient and emergency department of our hospital during 2013-2015;there is unreasonable drug use so that we need to strengthen management and standardize drug use.

Simultaneous determination of gallic acid and hesperidin contained in Xiaogu capsule by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To develop an HPLC method for simultaneous determination of gallic acid and hesperidin in Xiaogu capsule, in order to provide a simple, rapid and accurate method for quality control of the compound preparation of traditional Chinese medicine.</p><p><b>METHOD</b>Xiaogu capsule was extracted with methanol heating reflux method. Synergi 4 mu Hydro-RP 80A (4.6 mm x 250 mm, 5 microm) was adopted as the chromatographic column, with acetonitrile--0.04 mol x L(-1) phosphate monobasic sodium solution (20: 80) as the mobile phase. The flow rate was 1.0 mL x min(-1), the detection wavelength was 283 nm, and the column temperature was 25 degrees C.</p><p><b>RESULT</b>Under the conditions, gallic acid and hesperidin reached the baseline resolved peak, with a good linearity within the range of 21.6-216.0 mg x L(-1) (r = 0.999 93) for gallic acid, and 4.5-45.0 mg x L(-1) (r = 0.999 95) for hesperidin, respectively. Their average recoveries (n = 9) were 101.5% (RSD 3.7%) and 94.7% (RSD 2.7%), respectively. The average contents of gallic acid and hesperidin contained in Xiaogu capsule were detected to 5.10% and 0.091 1%, respectively.</p><p><b>CONCLUSION</b>The method established in this study can determine the content of gallic acid and hesperidin contained in Xiaogu capsule in a rapid and accurate manner, which provided reference for quality evaluation of the medicine.</p>

Effect of trihydroxybenzoic acid dimmer on induction of apoptosis of HL-60 cells / 药学学报

Trihydroxybenzoic acid dimmer is an anti-tumor compound which is separated from water-caltrop. This study is aimed to investigate its anti-proliferation effect on HL-60 cells and the possible mechanism of inducing apoptosis. MTT assay was used to test HL-60 cells proliferation. The apoptosis, ROS levels and the mitochondrial membrane potential were detected by flow cytometry. Cytochrome c released from mitochondria to cytosol fraction was detected by Western blotting. The activity of Caspase-9 and Caspase-3 were measured by colorimetric method. It was found to inhibit HL-60 cells proliferation in a dose and time-dependent manner. Compared with control group, it caused the increase of ROS levels and a concomitant dissipation of the mitochondrial membrane potential and induced cytosolic accumulation of cytochrome c and activities Caspase-9 and Caspase-3. Therefore it can be concluded that mitochondrial-dependent pathways was involved in its induction of apoptosis of HL-60 cells.

Molecular genetic analysis of mitochondrial DNA C1494T mutation in non-syndromic hearing loss of Chinese population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To conduct a molecular epidemiological survey on the mitochondrial DNA C1494T mutation in non-syndromic hearing loss patients in Chinese population.</p><p><b>METHODS</b>Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were used to screen the mitochondrial DNA 12S rRNA C1494T mutation in 20 patients with aminoglycoside antibiotic induced hearing loss, 136 sporadic non-syndromic hearing loss patients and 50 probands of pedigrees with non-syndromic hearing loss.</p><p><b>RESULTS</b>The C1494T mutation did not appear in all cases except for the positive control.</p><p><b>CONCLUSION</b>Incidence of mitochondrial DNA C1494T mutation is much lower than that of mitochondrial DNA A1555G mutation in non-syndromic hearing loss of Chinese population. Mitochondrial DNA C1494T mutation may be a rare variation in non-syndromic hearing loss and is not the main cause of aminoglycoside antibiotic induced-deafness.</p>

Phenotype heterogeneity associated with mitochondrial DNA A3243G mutation / 中国医学科学院学报

<p><b>OBJECTIVE</b>To discuss the clinical characteristics associated with mitochondrial DNA A3243G mutation.</p><p><b>METHODS</b>Clinical manifestations as well as results of brain CT and/or MRI scanning, blood level of lactic acid and muscle biopsy results of 25 mitochondrial encephalomyopathies patients whose A3243G mutations were analyzed.</p><p><b>RESULTS</b>Although all of the 25 patients carried mtDNA A3243G point mutation, their clinical manifestations varied greatly. Among them, there were 19 cases of mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes (MELAS), 2 cases of encephalopathies which could not be classified into any specific type, 2 cases of floppy infants, one case of Kearns-Sayer syndrome (KSS) and one case of mitochondrial entero-myopathy. Most patients showed abnormal cranial radiological findings and ragged-red-fibers on muscle biopsies. Elevation of blood lactic acid was notably found in all of the 25 patients.</p><p><b>CONCLUSIONS</b>Significant variations in clinical manifestation and brain images are the prominent features in patients with A3243G mutation. Mitochondrial diseases should be considered in patients with multiple organ involvement and elevated serum lactic acid mtDNA mutation examination is necessary for the diagnosis of mitochondrial diseases.</p>

Five single nucleotide polymorphisms of casein kinase I gamma 2 gene in children with familial febrile convulsions / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the association between single nucleotide polymorphisms (SNPs) of casein kinase I gamma 2 (CSNK1G2) gene and children with familial febrile convulsions.</p><p><b>METHODS</b>The study samples were collected from unrelated Chinese Han population of Hebei province, including a cohort of 53 children with familial febrile convulsions(FC) and a control cohort of 101 individuals. Genotypes of SNPs rs2074882, rs740423, rs2277737, rs4806825, rs1059684 were typed by polymerase chain reaction-restriction fragment length polymorphism.</p><p><b>RESULTS</b>The frequencies of the five SNPs complied well with the Hardy-Weinberg equilibrium in FC group and normal group. The distribution of genotype and frequencies of alleles of the SNPs rs740423, rs2277737, rs1059684 in familial febrile convulsions group was significantly different from that in control group. No significant difference was observed in the distribution of genotypes and frequencies of alleles at SNP rs2074882 between two groups. Analysis on rs4806825 was not made owing to its less allele frequency.</p><p><b>CONCLUSION</b>These data indicate that SNPs rs740423, rs2277737, rs1059684 of CSNK1G2 gene may contribute to familial febrile convulsions in children.</p>

Detection of mtDNA 11778 (G-->A) point mutation in a family with Leber's hereditary optic neuropathy by site-specific polymerase chain reaction / 中国医学科学院学报

<p><b>OBJECTIVE</b>To find a simple, fast, accurate, and quantitative PCR-based method for mutation detection, so as to identify mitochondrial DNA 11778 G-->A point mutation in patients with Leber's hereditary optic neuropathy (LHON).</p><p><b>METHOD</b>On the basis of sequencing of mtDNA from LHON proband, M primer for mutation and N primer for normal were designed to be coupled with reverse primer respectively. Specific PCRs were done on an amplifying condition with high stringency such as a well controlled annealing temperature, low Mg2+ concentration and less thermal cycles. The objective pedigree includes 10 individuals, were against 40 normal control persons.</p><p><b>RESULTS</b>Different ratios of indicative mtDNA 11778A-->G mutation were checked out from the proband, affected maternal members and a 10 year-old boy (up to now no appearance yet), whereas not appeared on normal spouses, paternal offsprings in the family, neither did on 40 controls.</p><p><b>CONCLUSION</b>This site-specific PCR method is a kind of general mutation analysis way, without the restriction of existence of endonuclease site. It can be applied for the gene diagnosis of known-mutation hereditary diseases such as LHON.</p>