Effect of moxibustion on M2 and P2X3 receptors of bladder tissue in rats with neurogenic bladder of detrusor areflexia after lumbar-sacral spinal cord injury / 中国针灸

OBJECTIVE@#To observe the effect of moxibustion at "Guanyuan" (CV 4) and "Shenque" (CV 8) on acetylcholine (Ach), adenosine triphosphate (ATP) and muscarinic-type choline receptor (M2) and purine receptor P2X3 in bladder tissue in the rats with neurogenic bladder (NB) of detrusor areflexia after lumbar-sacral spinal cord injury and explore the underlying mechanism of moxibustion for promoting detrusor contraction.@*METHODS@#Sixty SD rats were randomly divided into a model preparation group (n=45) and a sham-operation group (n=15). In the model preparation group, the modified Hassan Shaker spinal cord transection method was used to prepare the model of NB. In the sham-operation group, the spinal cord transection was not exerted except laminectomy and spinal cord exposure. Among the rats with successfully modeled, 30 rats were selected and divided randomly into a model group and a moxibustion group, with 15 rats in each one. On the 15th day after the operation, moxibustion was applied at "Guanyuan" (CV 4) and "Shenque" (CV 8) in the moxibustion group, 10 min at each acupoint, once a day. The consecutive 7-day treatment was as one course and the intervention for 2 courses was required. Urodynamic test was adopted to evaluate bladder function in rats. Using HE staining, the morphological changes in bladder tissue were observed. The content of Ach and ATP in bladder tissue was measured with biochemical method, and the protein and mRNA expression levels of M2 and P2X3 receptors in bladder tissue were detected with Western blot and real-time fluorescence quantification PCR method.@*RESULTS@#Compared with the sham-operation group, the maximum bladder capacity, leakage point pressure and bladder compliance were increased in the rats of the model group (P<0.05). Compared with the model group, the maximum bladder capacity, the leakage point pressure and bladder compliance were decreased in the rats of the moxibustion group (P<0.05). In the model group, the detrusor fibres were arranged irregularly, bladder epithelial tissues were not tightly connected and cell arrangement was disordered, combined with a large number of vacuolar cells. In the moxibustion group, compared with the model group, the detrusor fibres were arranged regularly, bladder epithelial cells were well distributed and vacuolar cells were reduced. Compared with the sham-operation group, the content of Ach and ATP in bladder tissue was decreased (P<0.05), the protein and mRNA expression levels of M2 and P2X3 receptors were reduced (P<0.05) in the model group. In the moxibustion group, the content of Ach and ATP in bladder tissue was increased (P<0.05) and the protein and mRNA expression levels of M2 and P2X3 receptors were increased (P<0.05) as compared with the model group.@*CONCLUSION@#Moxibustion at "Guanyuan" (CV 4) and "Shenque" (CV 8) may effectively improve bladder function in the rats with NB of detrusor areflexia after lumbar-sacral spinal cord injury and its underlying mechanism is related to promoting the release of Ach and up-regulating the expression of M2 receptor, thereby enhancing the release of ATP and increasing the expression of P2X3 receptor. Eventually, detrusor contraction is improved.

A chimeric influenza virus vaccine expressing fusion protein epitopes induces protection from human metapneumovirus challenge in mice.

Human metapneumovirus (HMPV) is a common virus associated with acute respiratory distress syndrome in pediatric patients. There are no HMPV vaccines or therapeutics that have been approved for prevention or treatment. In this study, we constructed a novel recombinant influenza virus carrying partial HMPV fusion protein (HMPV-F), termed rFLU-HMPV/F-NS, utilizing reverse genetics, which contained (HMPV-F) in the background of NS segments of influenza virus A/PuertoRico/8/34(PR8). The morphological characteristics of rFLU-HMPV/F-NS were consistent with the wild-type flu virus. Additionally, immunofluorescence results showed that fusion proteins in the chimeric rFLU-HMPV/F-NS could work well, and the virus could be stably passaged in SPF chicken embryos. Furthermore, intranasal immunization with rFLU-HMPV/F-NS in BALB/c mice induced robust humoral, mucosal and Th1-type dominant cellular immune responses in vivo. More importantly, we discovered that rFLU-HMPV/F-NS afforded significant protective efficacy against the wild-type HMPV and influenza virus challenge, with significantly attenuated pathological changes and reduced viral titers in the lung tissues of immunized mice. Collectively, these findings demonstrated that chimeric recombinant rFLU-HMPV/F-NS as a promising HMPV candidate vaccine has potentials for the development of HMPV vaccine.

Direct laser cooling the NH molecule with the pseudo-closed loop triplet-triplet transition including intervening electronic states.

Direct laser cooling molecule is useful way to obtain the accurate molecular spectroscopy. However, most of the reported direct laser cooling schemes are only involved the molecules with a singlet or doublet ground state because the one with a triplet ground state is more complex, especially when the first-excited state is not suitable for the pseudo-closed loop transition. Using NH as the prototype of the simplest heteronuclear molecule with a triplet ground state, we focus on constructing the direct laser cooling scheme with a pseudo-closed loop triplet-triplet transition including intervening electronic states. The potential energy curves and transition dipole moments are calculated for the X3Σ-, a1Δ, b1Σ+, and A3Π states by using the multireference configuration interaction including spin-orbit coupling with the aug-cc-pV5Z basis sets. The rotational and vibrational energy levels of each electronic state are obtained by solving the Schrödinger equation of nuclear motion with the obtained potential energy curves. A two-color laser cooling scheme is established based on the 3Π1 â†’ X3Σ- transition because the highly diagonal Franck-Condon factors make the transition suitable for constructing the pseudo-closed loop transition. The radiative lifetimes, the Doppler temperature, and the recoil temperature are calculated to access the cooling effect of the optical scheme. The results demonstrate that the 3Π1 â†’ X3Σ- transition is much superior to the other transitions and the intervening a1Δ and b1Σ+ will not significantly impact the pseudo-closed loop transition of the laser cooling scheme. The accumulate FCF reach 0.99996 implies that about 25,000 scattering photons are available before leaking, which can cool the NH molecule to the Doppler temperature of 20.2 µK.

Regulation of Hypnotic Active Components of Cnidii Fructus on Melatonin Synthesis Rate-limiting Enzyme AANAT in Rats with PCPA-induced Insomnia / 中国实验方剂学杂志

Objective:To observe the effects of Cnidii Fructus hypnotic active components （CHC） on the behaviors of rats with p-chlorophenylalanine （PCPA）-induced insomnia and melatonin （MT） synthesis rate-limiting enzyme arylalkylamine <italic>N</italic>-acetyltransferase （AANAT）， and explore the protective mechanism of CHC on the pineal gland. Method:Male SD rats of SPF grade were randomly divided into a blank control group， a model group， a MT group， and high-， medium-， and low-dose CHC groups with 10 rats in each group. Except for the blank control group， other groups received 4.5% PCPA suspension at 10 mL·kg^-1， intragastric administration， for two consecutive days. After PCPA model of insomnia was established， normal and model groups were gavaged at the same volume of 2% Tween-80， MT control group （10 mg·kg^-1）， CHC was high， medium and low （60， 30， 15 mg·kg^-1）， 10 mL·kg^-1， once a day， for consecutive 7 days. Four days after administration， open field， elevated cross maze， and pentobarbital sodium-induced sleep tests were conducted， respectively. Serum MT was detected by enzyme-linked immunosorbent assay. The mRNA expression level of AANAT was determined by real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR）. The expression of AANAT protein in the pineal gland was detected by Western blot. Result:Compared with the results in the blank control group， the total distance of open field activity and standing times and duration in the central area were increased （<italic>P</italic><0.05， <italic>P</italic><0.01）， the proportions of open arm entry （OE%） and open arm time （OT%） were decreased （<italic>P</italic><0.05）， and the sleep latency was prolonged （<italic>P</italic><0.01） in the model group. Compared with the model group， no significant difference was observed in the low-dose CHC group， while other groups exhibited reduced total distance of activity （<italic>P</italic><0.05， <italic>P</italic><0.01）， elevated OE% （<italic>P</italic><0.05）， shortened sleep latency， and prolonged sleep time （<italic>P</italic><0.05， <italic>P</italic><0.01）. Compared with the serum MT in the blank control group， that in the model group was decreased （<italic>P</italic><0.01）. Compared with the model group， no significant difference was observed in the low-dose CHC group， while other groups displayed increased serum MT （<italic>P</italic><0.05）. The mRNA and protein expression of AANAT was decreased in the model group as compared with that in the blank control group （<italic>P</italic><0.01）. Compared with the model group， the MT group and the high-dose CHC group showed up-regulated expression （<italic>P</italic><0.05）. Conclusion:CHC improved the behavioral indexes of PCPA-induced insomnia， increased the synthesis and secretion of MT in pineal cells， and elevated the serum MT level， which was related to the up-regulation of the mRNA and protein expression of AANAT in the pineal gland.

Effect of electro-acupuncture therapy on limb spasm and excitability of motor neurons in stroke rats / 浙江大学学报·医学版

To investigate the effect of electro-acupuncture therapy on limb spasm and excitability of motor neurons in stroke rats. Ischemic stroke model was induced with middle cerebral artery embolization in SD rats. Thirty-three modeled rats were randomly divided into model group, electro-acupuncture group, and baclofen group with 11 rats in each group, and another 10 rats were taken as sham operation group. The electro-acupuncture group and the baclofen group were treated with electro-acupuncture and baclofen tablets respectively. The model group and the sham operation group had no intervention. The neural function was evaluated with Bederson's scale and balance beam test; the muscle tension was measured with electrophysiography; the pathological changes of brain tissue was examined with HE staining; the content of glutamic acid (Glu) and γ-aminobutyric acid (GABA) in rat cerebral cortex was analyze with enzyme linked immunosorbent assay (ELISA) method, the expression of metabotropic glutamate receptor 1a () and γ-aminobutyric acid type B receptor subunit 1 () mRNA were detected with RT-qPCR. Compared with the model group, the neurological function scores of the electro-acupuncture group and the baclofen group showed a downward trend at d7 after operation (all >0.05), and the neurological function scores of the electro-acupuncture group and the baclofen group were significantly decreased at d12 after the operation (all 0.05). Compared with the model group, the electrophysiological results of the electro-acupuncture group and baclofen group were significantly increased after operation (all <0.05). The results of HE staining showed that there was no cell edema and degeneration in the sham operation group, no pyknosis of the nucleus, and no bleeding in the interstitium. Cell edema and degeneration and mesenchymal congestion appeared in the model group. Compared with the model group, the cytoplasmic edema and degeneration and the interstitial bleeding in the electroacupuncture group and the baclofen group were reduced. Compared with sham operation group, the Glu content and the relative expression of mRNA was increased in the model group, electro-acupuncture group and baclofen group, while the GABA content and the relative expression of mRNA decreased (all <0.05). Compared with model group, the Glu content and the relative expression of mRNA in the electro-acupuncture group and baclofen group decreased, and the GABA content and relative expression of mRNA increased (all <0.05). Electro-acupuncture may improve limb spasm after stroke through regulating the expression of Glu and GABA in the cerebral cortex and the excitability of motor neurons in rats.

Deletional variant of REEP1 gene in a pedigree affected with spastic paraplegia type 31 / 中华医学遗传学杂志

OBJECTIVE@#To detect pathogenic variation in a pedigree affected with hereditary spastic paraplegia type 31 and explore its molecular pathogenesis.@*METHODS@#Customized Roche NimbleGen capture probes were used to capture all exons of the target genes in relation with hereditary spastic paraplegia. The DNA samples were also assayed with fluorescent quantitative PCR as well as chromosomal microarray analysis using CytoScan HD chip.@*RESULTS@#The proband and her father and grandfather were found to carry a deletion for position 85 992 693-86 842 693 on chromosome 2, which spanned approximately 900 kb and encompassed the REEP1 gene. The latter has been specifically associated with hereditary spastic paraplegia type 31. The same deletion was not found in her mother who is phenotypically normal.@*CONCLUSION@#The deletional variation of the REEP1 gene probably underlies the disease in this pedigree.

Association of Adolescent Obesity with Nonalcoholic Fatty Liver Disease and Related Risk Factors in Xi 'an, China.

INTRODUCTION AND AIM: To investigate the association of adolescent obesity with nonalcoholic fatty liver disease (NAFLD) and related risk factors in Xi'an, China. MATERIAL AND METHODS: A total of 4141 adolescents (2,061 girls and 2,080 boys, mean age: 18.62 ± 0.66 years, age range 15-22 years) were enrolled in this investigation. Anthropometric index was measured, and liver ultrasonography and liver function biochemical test were performed in all the subjects. T test, χ2 test and logistic regression analysis were used for statistical analyses. RESULTS: The total rates of obesity was 7.9% (08/4,141). The prevalence rate of NAFLD was 8.1% (335/4141) with a declining trend from obesity, overweight to normal BMI. NAFLD prevalence rate in obese boys was significantly higher than in girls (χ2 = 56.5, P < 0.01). BMI, body weight, ALT, and AST in NAFLD group were higher than in non-NAFLD group (P < 0.05). The tangent point of ALT was 36 U/L using Youden index in boys, and 33 U/L in girls. CONCLUSION: The prevalence of obesity and NAFLD in adolescents is higher in Xi'an than anticipated. Body weight and BMI may be the associated independent risk factors of NAFLD.

Novel FOXG1 mutation in a patient with congenital Rett variant: a case report / 天津医药

@#Objective To summarize the clinical features of children with congenital Rett variant caused by mutation of FOXG1 and provide the reference for the diagnosis and treatment of the disease. Methods The clinical data of a patient diagnosed as congenital Rett syndrome variant type were summarized. The DNA samples of peripheral blood from the patient and her parents were extracted. The targeted high-throughput sequencing technology was used to detect the sequence of targeted genes, which were associated with the symptoms of the child. Genes were then verified by sanger sequencing. Chromosomal microarray analysis was performed to detect chromosome microdeletions and microduplications. Results The child carried the c.506dupG, p.G169Gfs＊ 286 heterozygous mutations on FOXG1 gene, which located in 14q12, and her parents were wild-type. After querying the HGMD, Clinvar and dbSNP databases, we found that it was not reported. This case was clearly diagnosed as congenital Rett syndrome variant type. We confirmed that the mutation locus was a new mutation. Conclusion For cases with congenital Rett variant manifestations, FOXG1 gene mutation examination is recommended, and preventive treatment of partially predictable dysfunction should be carried out.

The Application of 256-Slice Spiral CT in the Diagnosis of Obstructive Sleep Apnea Hypopnea Syndrome / 昆明医科大学学报

Objective To measure the upper airway of obstructive sleep apnea hypopnea syndrome by 256 slice spiril CT, and to access the airway obstruction plane with the airway plane data in OSAHS patients.Through these measurements, we can provide assistance for clinical diagnosis and treatment.Methods This study randomly selected 178 patients diagnosed with OSAHS and 110 cases non-snoring healthy people as the control group by the Philips 256 slice CT.Under the nasopharyngeal area, velopharyngeal area, glossopharyngeal area, epiglottis area, we measured the cross-sectional area and volume of the narrowest plane in the two groups of quiet respiration and Müller status.Results The OSAHS group underwent quiet respiration and Müller status during CT scanning, and the two states about epiglottis area in cross-sectional area and volume had no significant difference, Other groups had differences between the parameters.There was a significant difference in the volume about velopharyngeal area and glossopharyngeal area. In the control group undergoing quiet respiration and Müller status during CT scanning, there was difference in velopharyngeal cross-sectional area.Other parameters had no significant difference.Conclusions The obstruction plate of OSAHS patients with 256-slice spiril CT measurement is mostly in the velopharyngeal area and glossopharyngeal area. The volume measurement of upper airway by CT can predict airway obstruction plate in patients with OSAHS.

Training and application of standardized patients in otolaryngology / 基础医学与临床

Objective To investigate the application of standardized patients in the department of otolaryngology. Methods Two otolaryngology scripts were designed and four standardized patients were trained. Ten students were offered standardized patients teaching twice. Students' and clinical teachers' feedback, as well as objective grading for each student were analyzed. Results The performance scores for all the standardized patients were o-ver 4 points(1-5 points). Standardized patients and clinical teachers have a high degree of consistency in grad-ing the students. 70%(7/10) of the students considered standardized patients' immediate comment as the most benefits. Conclusions Well trained standardized patients are competent for the role of an actor,evaluator and in-structor in teaching and assessing students. The standardized patients as a teaching component should be promoted in otolaryngology.

Metabolic study of Mori Cortex on nephrogenic edema in rats / 中成药

AIM To conduct a metabolic research for a better understanding of nephrogenic edema and to assess the integral efficacy of Mori Cortex in rat model.METHODS The serum creatinine,urea nitrogen,albumin and urinary protein levels in rats were detected.UPLC-QTOF-MS was used to detect the urine metabolites changes,Principal Component Analysis (PCA) and Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA)were used to screen potential biomarkers,after whose quantification,Mev software was adopted for heat map draw-ing and hierarchical cluster analysis.RESULTS The model rats manifested significantly increased levels of serum creatinine,urea nitrogen and urinary protein,decreased albumin level,and an obviously excessive amino acid metabolism as well.The 41 identified biomarkers were mainly related to disturbances in phenylalanine,pyrimidine,arginine and proline,glycine,serine and threonine,tryptophan,cysteine and methionine metabolism,and biosynthesis of pantothenic acid and coenzyme A.A reversal trend in aforementioned levels of biochemical indexes and most biomarkers due to the intervention by Mori Cortex signaled an improvement in the metabolic disorder,renal dysfunction and edema.CONCLUSION The metabolic study demonstrates the pathological status of nephrogenic edema and assesses the effect of Mori Cortex from an overall perspective,highlighting a new approach for illustrating Chinese medical syndrome and the underlying mechanism in the management of traditional Chinese medicine.

Simultaneous Determination of 7 Components in Kangjun Xiaoyan Tablets by RP-HPLC / 中国药房

OBJECTIVE:To develop a method for simultaneous determination of chlorogenic acid,baicalin,aloe emodin,rhe-in,emodin,chrysophanol and physcione in Kangjun xiaoyan tablets. METHODS:RP-HPLC method was adopted. The determina-tion was performed on Agilent Extend C18 with mobile phase consisted of acetonitrile-0.5% phosohoric acid solution(gradient elu-tion)at the flow rate of 1.0 mL/min. The detection wavelengths were set at 327 nm(chlorogenic acid),280 nm(baicalin)and 450 nm(aloe emodin,rhein,emodin,chrysophanol,physcione). The column temperature was 30℃,and sample size was 10μL. RE-SULTS:The linear ranges of chlorogenic acid,baicalin,aloe emodin,rhein,emodin,chrysophanol and physcione were 0.2094-4.1880μg(r=0.9999),0.3720-7.4400 μg(r=0.9999),0.0063-0.1262 μg(r=0.9999),0.0116-0.2322 μg(r=0.9999),0.0053-0.1060 μg(r=0.9998),0.0128-0.2564 μg(r=0.9999),0.0165-0.3303 μg(r=0.9998),respectively. The limits of quantita-tion were 2.01,1.93,0.76,1.25,1.24,0.53 and 1.53 ng;the limits of detection were 0.98,0.65,0.25,0.42,0.41,0.18 and 0.52 ng,respectively. The recoveries were 98.41%-100.40%(RSD=0.76%,n=9),96.17%-100.10%(RSD=1.58%,n=9), 96.11%-100.10%(RSD=1.33%,n=9),96.29%-100.80%(RSD=1.85%,n=9),96.88%-100.10%(RSD=1.22%,n=9), 97.81%-101.90%(RSD=1.64%,n=9),96.46%-101.30%(RSD=1.85%,n=9),respectively. CONCLUSIONS:The method is accurate,reliable and reproducible. It can be used for simultaneous determination of 7 components in Kangjun xiaoyan tablets.

Main chemical components and pharmacological effects of Garcinia mangostana L.:research advances / 国际药学研究杂志

Garcinia mangostana L. is a common tropical plant,which is also called mangosteen. Its exocarp is used as a tradi?tional medicine in India,Thailand,Myammar and other Southeast Asian countries. In recent years,systematic studies on the chemi?cal constituents of the plant have shown that G. mangostana contains various xanthone compounds in a large amount. Many studies have shown that these compounds display many pharmacological activities,such as antioxidant,antiproliferative,pro-apoptosis,anti-inflammation and antibacterial activities,which has attracted great attention in the medical community. This paper reviews the main chemical constituents of G. mangostana and their pharmacological effects.

Inlfuence on Substance and Energy Metabolism of Diabetic Mouse Model of Chemical Split Fractions ofMori Cortex / 世界科学技术-中医药现代化

This article was aimed to study the impact on substance and energy metabolism by chemical split fractions of Mori Cortex among hypoglycemic diabetic mouse model, in order to explain the new hypothesis of the science connotation in nature and flavor of traditional Chinese medicine (TCM). Male Kunming mice were intraperitoneally injected with a large dose of streptozotocin (STZ) (170 mg·kg-1) to establish type 1 diabetes mellitus mouse model. Medication was given consecutively for four weeks. The enzyme-linked immunesorbent assay (ELISA) was used to detect glucosekinase (GCK), glycogen phosphorylase (PYGL), pyruvate dehydrogenase (PDH), α-ketoglutarate dehydrogenase (α-KGDHC), phosphoglycerate kinase (PGK), acetyl coenzyme A (acetyl-CoA), adenylate kinase (ADK), fumarase (FUM), cytochrome C reductase (CCR), cytochrome C oxidase (COX) and other indicators. Enzymatic detection was used to determine the content of ATP coenzyme (ATPs), the content and ratio of NAD and NADH, the content of myocardial cell Na+-K+ ATP enzyme, as well as the content of ATP and ADP. The results showed that in the model group, the expression of PYGL was increased; and the expressions of GCK and PDH were decreased. It prompted that the source of glucose increased and the expelling of glucose decreased. The glucose level was increased. The COX expression was reduced and the respiratory chain was blocked. It regulated oxidative phosphorylation and the substrate phosphorylation level. It upregulated the expression of CCR, ATPs, NAD+, PGK, α-KGDHC and ADK. However, the expression of FUM was decreased. The activity of Na+-K+ ATPase was decreased significantly. At last, the metabolic disorders appeared. Mori Cortex aqueous extracts and the chemical split fractions significantly increased the GCK and PDH level in substance metabolism among diabetic mice. The levels of PYGL, α-KGDHC, PGK and acetyl-CoA were decreased (P < 0.05, or P <0.01). Meanwhile, it increased ATP and FUM, myocardial cell Na+-K+ ATP enzyme, and COX level in the energy metabolism (P < 0.05). It decreased the level of NAD+, CCR and ATPs (P < 0.05, or P <0.01). It was concluded that both the aqueous extracts and chemical split fractions of Mori Cortex can effectively improve the substance and energy metabolism disorders of diabetic mouse model. This effect may be related to the cold nature of Mori Cortex.

Effect of curcumin on aged Drosophila melanogaster: a pathway prediction analysis / 中国结合医学杂志

<p><b>OBJECTIVE</b>To re-analyze the data published in order to explore plausible biological pathways that can be used to explain the anti-aging effect of curcumin.</p><p><b>METHODS</b>Microarray data generated from other study aiming to investigate effect of curcumin on extending lifespan of Drosophila melanogaster were further used for pathway prediction analysis. The differentially expressed genes were identified by using GeneSpring GX with a criterion of 3.0-fold change. Two Cytoscape plugins including BisoGenet and molecular complex detection (MCODE) were used to establish the protein-protein interaction (PPI) network based upon differential genes in order to detect highly connected regions. The function annotation clustering tool of Database for Annotation, Visualization and Integrated Discovery (DAVID) was used for pathway analysis.</p><p><b>RESULTS</b>A total of 87 genes expressed differentially in D. melanogaster melanogaster treated with curcumin were identified, among which 50 were up-regulated significantly and 37 were remarkably down-regulated in D. melanogaster melanogaster treated with curcumin. Based upon these differential genes, PPI network was constructed with 1,082 nodes and 2,412 edges. Five highly connected regions in PPI networks were detected by MCODE algorithm, suggesting anti-aging effect of curcumin may be underlined through five different pathways including Notch signaling pathway, basal transcription factors, cell cycle regulation, ribosome, Wnt signaling pathway, and p53 pathway.</p><p><b>CONCLUSION</b>Genes and their associated pathways in D. melanogaster melanogaster treated with anti-aging agent curcumin were identified using PPI network and MCODE algorithm, suggesting that curcumin may be developed as an alternative therapeutic medicine for treating aging-associated diseases.</p>

Growth Inhibition of Multiple Myeloma Cells Caused by MicroRNA-15a and Its Mechanisms / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effect and possible mechanisms of miR-15a on growth of multiple myeloma(MM) cells.</p><p><b>METHODS</b>MM cell lines (U266 and RPMI8226) were transfected by lentiviral particles. MM stable cell lines were selected and collected by flow cytometry (FCM). Proliferation of MM cells before and after miR-15a high expression was detected by CCK-8 method. Apoptosis of MM cells before and after miR-15a high expression was detected by AO/EB dying, Hoechst 33258 dying and FCM, respectively. Cell cycle of MM cells before and after miR-15a high expression was detected by FCM. The expressions of miR-15a, BMI-1 and BCL-2 mRNA of MM cells before and after miR-15a high expression were detected by real-time PCR. The expressions of BMI-1 protein of MM cells before and after miR-15a high expression were detected by Western blot.</p><p><b>RESULTS</b>MM stable cell lines with miR-15a high expression was acquired. CCK-8 result showed that high expression of miR-15a could inhibit growth of MM cells (U266 and RPMI8226). AO/EB dying, Hoechst 33258 dying and FCM testing results showed that high expression of miR-15a could significantly induce apoptosis of MM cells (U266 and RPMI8226). The apoptosis rates of U266 and RPMI8226 cells in high expression group and control group were 90.52% vs 37.08% and 59.40% vs 44.17%, respectively. Meanwhile, FCM testing results showed that high expression of miR-15a could induce G1 arrest of MM cells (U266 and RPMI8226), which proportion of G1 phase were 41.50%±0.64%, 45.31%±0.77%, respectively. Real-time PCR results showed that during the growth inhibition process of MM cells caused by miR-15a high expression, the expression of BCL-2 mRNA decreased, but there was no significant changes in the expression of BMI-1 mRNA, while the expression of BMI-1 protein decreased significantly.</p><p><b>CONCLUSION</b>High expression of miR-15a can induce cell cycle arrest and apoptosis of MM cells, then inhibit their growth. The mechanisms may be related with the negative regulation of BMI-1 and BCL-2 genes in post-transcription level caused by miR-15a.</p>

Expression and Clinical Significance of Bmi-1 and P14 in Extranodal NK/T-cell Lymphoma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>This study was aimed to investigate the expression and clinical significance of Bmi-1 and P14 in extranodal NK/T-cell lymphoma (ENKTCL) tissue.</p><p><b>METHODS</b>Maxvision immunohistochemistry technique was used to detect the expression level of Bmi-1 and P14 in the tissues of 21 patients with ENKTCL and 11 normal lymph nodes. The correlation of Bmi-1 or P14 expression with the clinical features and the correlation between Bmi-1 and P14 expression were analyzed.</p><p><b>RESULTS</b>The expression of Bmi-1 protein was higher in tissues of ENKTCL than that in tissues of lymph nodes, and the Bmi-1 expression levels did not correlate with patients' sex, age, lactate dehydrogenase (LDH), International Prognostic Index (IPI) scores and B symptoms (P > 0.05), except for clinical stage (P < 0.05). The P14 protein expression level was lower in ENKTCL tissues than in normal lymph node tissues, which did not correlate with age, sex, LDH, IPI scores, clinical stage and B symptoms. Correlation test showed a negative correlation between Bmi-1 and P14 (r = -0.472, P = 0.031).</p><p><b>CONCLUSION</b>Bmi-1 protein over-expresses in ENKTCL tissues that may display a negative-regulation effect on P14 in the genesis and progress of ENKTCL.</p>

Analysis on chemical constituents in Danggui-Shaoyao-San by LC-Q-TOF-MS and LC-IT-MSn / 中草药

Objective: To study the constituents in Danggui-Shaoyao-San (DSS, a powder prescription with Angelicae Sinensis Radix, Paeoniae Radix Alba, Atractylodis Macrocephalae Rhizoma, Poria, Alismatis Rhizome, and Chuanxiong Rhizome). Methods: To analyze the 70% ethanol-water extract from DDS by LC-Q-TOF-MS and LC-IT-MS. The analysis was performed on an Agilent Zorbax SB-C18 column and Rapid Resolution HT column (50 mm × 4.6 mm, 1.8 μm) with gradient elution of 0.05% formic acid (A)-acetonitrile (B) at the flow rate of 0.6 mL/min. The constituents from DSS were identified according to the relative molecule weight by LC-Q-TOF-MS and multistage MS informatics by LC-IT-MS. All the peaks were assigned to the corresponding traditional Chinese medicinal materials. Results: Thirty compounds from DDS could be identified, and 14 of them were identified for the first time in DSS including six organic acids, such as chlorogenic acid (8) and galloylsucrose (5-7); two protostane trierpenoids, such as 16-oxoalisol A 23-acetate (22); four cyclopeptides, such as cyclo tetraleucyl (or isoleucyl) (14) and other types of compounds, such as N-(1-deoxy-D-fructos-1-yl) pyroglutamic acid (1) and adenosine (3). Conclusion: This work investigates the compounds from DSS, and the results could provide the evidence for the research on active constituents and quality control of DSS.

A new phthalide from angelicae sinensis radix / 中国中药杂志

Angelicae Sinensis Radix, dried roots of Angelicae sinensis, is a well-known traditional Chinese medicine. Six compounds were isolated and purified by silica gel and preparative HPLC. According to physicochemical properties and spectral data, the compounds were identified as senkyunolide H-7-acetate (1), o-phthalic acid (2), diisobutyl phthalate (3), p-hydroxyphenylethanol ferulate (4), ferulic acid (5) and coniferylferulate (6). Compound 1 was a new one.

Anti-leukemia mechanism of miR-17 and miR-20a silencing mediated by miRNA sponge / 中国实验血液学杂志

This study was aimed to quantitatively detect the expression levels of pre-miR-17 and pre-miR-20a in acute leukemia patients and eight kinds of leukemia cell lines, and to investigate the anti-leukemia mechanism of miR-17 and miR-20a silence mediated by miRNA Sponge. Quantitative real-time PCR was used to detect the mRNA expression levels of pre-miR-17 and pre-miR-20a in patients with various types of leukemia and leukemia cell lines. The Jurkat cells over-expressing miR-17 and miR-20a were transfected with recombinant lentivirus-transfecting units targeted at miR-17 and miR-20a plus 6 µg/ml of polybrene. Then the proliferation ability and cell cycle of Jurkat cells was evaluated by CCK-8 and flow cytometry respectively. The results showed that the expression level of pre-miR-17 and pre-miR-20a in all leukemia patients was significantly higher than that in normal group(P < 0.05), the expression of pre-miR-17 and pre-miR-20a in acute lymphoid leukemia was significantly higher than that in acute myeloid leukemia(P < 0.05), and the pre-miR-17 and pre-miR-20a expression level did not correlate significantly with high white blood cell count>20.0×10(9)/L(P > 0.05). The miR-17 and miR-20a silencing mediated by miRNA Sponge led to a significant decrease of cell growth, restored G1 accumulation and increase of cell apoptosis. It is concluded that the expression of miR-17 and miR-20a is upregulated in leukemia patients, which may contribute to leukemogenesis. Over-expressed miR-17 and miR-20a promote cell growth and cell cycle progression, and inhibit apoptosis through negatively-regulating P21 and E2F1 after-transcriptionally.