Establishment of gastric cancer SGC-7901 cell line with stable lncRNA BC002811 silencing / 医学研究生学报

Objective The aim of this study was to construct a recombinant lentivirus-mediated short hairpin RNA (shRNA) expression vector targeting the long non-coding RNA (lncRNA) BC002811 and establish a gastric cancer SGC-7901 cell line with stable BC002811 down-regulation. Methods Three small interfering RNAs (siRNA) were designed and synthesized. The best sequence for RNA interference was selected by real-time quantitative PCR (qPCR) and inserted into the lentiviral vector pLVX-shRNA2. After identification by DNA sequencing, the lentiviral vectors carrying BC002811 shRNA were packaged in HEK293T cells. The lentiviral particles were collected to infect human gastric cancer SGC-7901 cells. After screened by limiting dilution analysis, the SGC-7901 cell line with stable BC002811 down-regulation was established, the expression level of BC002811 detected by qPCR, and the effect of BC002811 on the proliferation of the cells analyzed by MTS. Results The results of qPCR showed that BC002811 siRNA-1 was the most effective siRNA sequence, with a knockdown efficiency of 87%. The recombinant lentiviral vector was packaged in the HEK293T cells with a viral titer of 3.7 × 108 TU/mL in the shRNA-1 group as compared with 4.5 × 108 TU/mL in the control. The expression of BC002811 in the shRNA-1 group was only 10% of that in the control group (P < 0.01), which indicated the successful establishment of the gastric cancer SGC-7901 cell line with stable BC002811 down-regulation. BC002811 knockdown significantly inhibited the proliferation of the SGC-7901 cells in the shRNA-1 group as compared with the control. Conclusion A recombinant lentiviral vector expressing BC002811 shRNA was successfully constructed and the gastric cancer cell line SGC-7901 with stable BC002811 silencing was established.

Cloning and expression analysis of cycloartenol synthase gene in Fritillaria cirrhosa / 中草药

Objective To obtain the key enzyme gene involved in alkaloids biosynthesis pathway of Fritillaria cirrhosa, cycloartenol synthase (CAS) gene was cloned, and its bioinformatics analysis and gene expression pattern were also performed. Methods The open reading frame (ORF) of F. cirrhosa CAS (FcCAS) was amplified by PCR based on transcriptome sequencing. The bioinformatics analysis of FcCAS cDNA sequences was carried out by some online tools. Meanwhile, using real-time quantitative PCR (qRT-PCR) to analyze the gene expression patterns between wild and regeneration bulbs. Moreover, the content of total alkaloids in wild and regenerated bulbs were also investigated. Results The results showed that CAS had a length of 2 271 bp ORF, which encoding 756 amino acids. The phylogenetic tree analysis showed that FcCAS were highly similar to the corresponding proteins in Asparagus officinalis, Musa acuminate, and Elaeis gunineensis from the NCBI website, and the similarities were more than 80%. The results of qRT-PCR and total alkaloids assay showed that the changing trend of the expression level of FcCAS was consistent with that of the content of total alkaloids, and higher alkaloid accumulation was in regeneration bulbs than wild bulbs. Conclusion The expression of FcCAS gene varied widely in different tissues. These findings suggested that FcCAS was a biological functional protein induced by hormone combination, which laid a theoretical foundation for the improvement of the alkaloid content by using the genetic engineering.

Determination of six constituents in Shiyifang Vinum by HPLC / 中成药

AIM To establish an HPLC method for the content determination of six constituents in Shiyifang Vinum (Notoginseng Radix et Rhizoma,Dipsaci Radix,Carthami Flos,etc.).METHODS The content determination of notoginsenoside R1,ginsenoside Rg1,ginsenoside Rb1 and asperosaponin Ⅵ was performed on a 30℃ thermostatic Inertsil(R) ODS-3 C18column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1％ phosphoric acid flowing at 1.0 mL/min in a gradient manner,and the detection wavelength was set at 203 nm.The content determination of brucine and strychnine was conducted on a 30 ℃ thermostatic Geminni(R) C18 110(A) column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-mixed solution of 0.01 mol/L sodium heptanesulfonate and 0.02 mol/L potassium dihydrogen phosphate flowing at 1.0 mL/min in an isocratic elution manner,and the detection wavelength was set at 260 nm.RESULTS Six constituents showed good linear relationships within their own ranges (r ＞ 0.999 0),whose average recoveries were 98.52％-99.96％ with the RSDs of 2.0％-2.3％.CONCLUSION This simple,accurate and reproducible method can be used for the quality control of Shiyifang Vinum.

Exploration on extraction and enrichment process of anti-inflammatoryand analgesic active-fraction(ARF)which containsmethyl salicylate glycosides in Dianbaizhu / 国际药学研究杂志

Objective The active ingredient was used as index to optimize the extraction and enrichment process of anti-in-flammatory and analgesic active-fraction(ARF)of Dianbaizhu. Methods Methyl salicylate triglycoside-B was chosen as index com-ponent to extract and enrich methyl salicylate glycosides. Extraction and elution solvents were optimized. The HPLC fingerprint was ob-tained with Thermo Hypersil Gold C18(250 mm×4.6 mm,5μm)column and a gradient elution with the mobile phase consisting of ace-tonitrile(A)-0.2％acetic acid(B)at a flow rate of 1.0 ml/min. And the detection wavelength was set at 294 nm. Results The opti-mized extraction solvent of Dianbaizhu was the 30％ethanol and the optimized elution solvent of ARF enriched by AB-8 macroporous resins was the 35％ethanol. The methodological study on similarity and RSD in ARF HPLC fingerprint of three batches of samples cor-responded to related regulations. Conclusion The extraction and enrichment process of ARF is stable and repeatable.

Magnetic resonance imaging characteristics of unilateral versus bilateral intraplaque hemorrhage in patients with carotid atherosclerotic plaques / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the difference in the vulnerability of carotid atherosclerotic plaques in patients with unilateral and bilateral intraplaque hemorrhage (IPH).</p><p><b>METHODS</b>A retrospective analysis was conducted among 44 patients with unilateral IPH (30 cases) or bilateral IPH (14 cases) in the carotid plaques detected by magnetic resonance imaging (MRI) in our hospital between December, 2009 and December, 2012. The age, maximum wall thickness and incidence of fibrous cap rupture were compared between the two groups.</p><p><b>RESULTS</b>Compared with those with unilateral IPH, the patients with bilateral IPHs had a significantly younger age (66.6∓9.4 years vs 73.7∓9.0 years, P=0.027), a significantly greater maximum plaque thickness (6.3∓1.9 mm vs 5.0∓1.3 mm, P=0.035) and a higher incidence of ulcers (50% vs 13.3%, P=0.025). Logistic regression analysis revealed a significant association between bilateral IPHs and the occurrence of ulcer with an odd ratio (OR) of 6.5 (95% confidence interval [CI]: 1.5-28.7, P=0.014). After adjustment for gender in Model 1, bilateral IPHs were still significantly associated with presence of ulcer (OR=5.7, 95%CI: 1.1-29.2, P=0.036). But after adjustment for age (P=0.131) or maximum plaque thickness (P=0.139) in model 2, no significant correlation was found between bilateral IPHs and the presence of ulcer.</p><p><b>CONCLUSION</b>Compared with patients with unilateral IPH, those with bilateral IPHs are at a younger age and have a greater plaque burden and a higher incidence of fibrous cap rupture, suggesting a greater vulnerability of the carotid plaques in patients with bilateral IPHs.</p>

Cycle arrest of prostate carcinoma DU-145 cells induced by pseudolaric acid B / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of pseudolaric acid B (PLAB) on cell proliferation and cycle of human prostate carcinoma DU-145 cells. method: Its inhibitory effect on the cell growth was measured by MTT method. Characteristics of cell death were determined by Hoechest 33342 staining. The cell cycle was detected by flow cytometry. The expressions of cyclin B1, cyclin D1 and CDK1 were detected by Real time-PCR and Western blot, respectively.</p><p><b>RESULT</b>PLAB notably inhibited DU-145 cell growth in a dose- and time dependent manner (P < 0.05). Its IC50 values of PLAB for DU-145 cells for 24, 48 and 72 h were 4.53, 2.39 and 2.08 micromol x L(-1), respectively. Having been treated with 5 micromol x L(-1) PLAB for 24 h, the cells showed such apoptosis characteristics as nuclei chromatin condensation and apoptotic body. With the increase in PLAB concentration, the proportion of G2/M phase cells strikingly increased in a dose- and time dependent manner (P < 0.05), meanwhile cyclin B1 and CDK1 showed over-expressions (P < 0.05), and the cyclin D1 showed under-expression (P < 0.05).</p><p><b>CONCLUSION</b>PLAB can inhibit the growth of DU-145 cells and induce the cell cycle G2/M arrest, accompanied with the over-expression of cyclin B1 and CDK1, which may be related with its regulation cycle-associated protein degradation.</p>

Pathology of myocardial fatty infiltration: an autopsy report from 117 cases / 中华心血管病杂志

<p><b>OBJECTIVE</b>To explore the risk factors related to the formation of myocardial fatty infiltration and possible pathological consequences.</p><p><b>METHODS</b>The macroscopic and microscopic findings in 117 autopsy cases with myocardial fatty infiltration were examined during October, 2001 to June, 2009.</p><p><b>RESULTS</b>There was a significant positive correlation between the macroscopic grading of subepicardial adipose tissue and the microscopic myocardial fatty infiltrative degree(r(s) = 0.57, P < 0.01) but there was no correlations between the myocardial fatty infiltrative degree and age as well as coronary arteriosclerosis (all P > 0.05). The percent of myocardial atrophy was 39.32% (46/117), and the rate of myocardial atrophy in mild myocardial fatty infiltration group (13/63, 20.63%) was significantly lower than that in moderate myocardial fatty infiltration group (22/39, 34.92%; chi(2) = 12.14, P < 0.01) and in severe myocardial fatty infiltration group (11/15, 73.33%; chi(2) = 13.42, P < 0.01). There were 28 sudden cardiac deaths among the 117 cases including 6 deaths due to myocardial fatty infiltration.</p><p><b>CONCLUSIONS</b>Myocardial fatty infiltration is often associated with myocardial atrophy, even with sudden cardiac death but is not an accompanying pathologic changes of aging and coronary arteriosclerosis.</p>

Influence of celecoxib on invasiveness of human high-metastatic nasopharyngeal carcinoma cell line CNE-2Z / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the effect and mechanism (a selective cyclooxygenase-2 inhibitor) on invasive ability of human nasopharyngeal carcinoma (NPC) line CNE-2Z.</p><p><b>METHODS</b>The proliferation of NPC cells was examined by MTT assay. The invasive and migrating ability of NPC cells was detected with transwell chamber. E-cadherin protein expression was detected by immunocytochemistry and the expressions of Cox-2 and E-cadherin mRNA were analyzed by reverse transcription-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>MTT showed that celecoxib inhibited CNE-2Z proliferation in dose-dependent manner, the survival rate of cells treated with 25, 50, 100 µmol/L celecoxib (x(-) ± s) for 24 h was (94.75 ± 1.34)%, (91.77 ± 2.70)%, (64.54 ± 1.20)%, respectively, and the survival rate of cells treated for 48 h was (88.41 ± 1.28)%, (78.84 ± 1.56)%, (52.46 ± 2.25)%, respectively, the concentration of 50% inhibition concentration of a substance (IC50) was 100 µmol/L, the difference was statistically significant between different concentration groups in the same time-point (respectively, F were 462.204 and 1328.306, P < 0.01). Treated with different concentrations of celecoxib (0, 25, 50 µmol/L) for 24, the cell numbers (x(-) ± s) through PVPF by tumor invasion assay were (263.7 ± 13.5), (185.3 ± 8.7) and (144.0 ± 8.2), the difference was statistically significant between the experimental and control group (F = 102.089, P < 0.01). Immunocytochemistry showed that celecoxib significantly induced the increase of E-cadherin protein expression, also with a dose-dependence in 0 µmol/L, 25 µmol/L, 50 µmol/L group was (21.7 ± 2.6), (28.7 ± 2.4), (40.3 ± 1.3), and 50 µmol/L group increased significantly (F = 78.637, P < 0.01). RT-PCR showed that celecoxib reduced the expression of Cox-2 mRNA expression in 25, 50 µmol/L group decreased significantly compared with the control group (respectively, t were 23.950 and 36.651, P < 0.01), but it enhanced the expression of E-cadherin mRNA expression in 25, 50 µmol/L group was significantly higher (respectively, t were 35.829 and 81.497, P < 0.01).</p><p><b>CONCLUSION</b>Celecoxib can inhibits the invasive ability of NPC cell line CNE-2Z, which possibly relates with the upregulated expression of E-cadherin.</p>

Histopathological characterization of drug-related death / 法医学杂志

OBJECTIVE@#To study the histopathological changes in drug-related death cases in order to provide valuable information for its diagnosis.@*METHODS@#Thirty cases of drug-related death were collected for systemic autopsy and histopathology examination. Ante mortem history and other informations of each case were also reviewed and analyzed.@*RESULTS@#Injection marks, emaciation, asphyxia and histopathological changes in critical organs and tissues correlated with addiction behavior. In the 30 cases, 20% died of diseases, 33.3% acute drug intoxication, 26.7% quitting drug, 10% sudden death, and 10% outside violence.@*CONCLUSION@#Systemic autopsy and histopathology examination in drug-related death are useful for determination of the cause of death in these cases.

Pathology of accidental electrocution: an autopsy study of 16 cases / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the pathologic findings seen in lethal cases due to accidental electrocution.</p><p><b>METHODS</b>The macroscopic and microscopic findings in 16 autopsy cases died of electrocution encountered during the period from January, 2001 to July, 2008 were retrospectively reviewed.</p><p><b>RESULTS</b>Typical electric marks were found on gross examination in 5 of the 16 cases studied. Histologically, 11 of the 16 cases showed evidence of electric burn. The morphologic features of atypical electric marks varied. Simple epidermal exfoliation and color changes were relatively common. Pathologic changes in internal viscera included disarray of myocardial fibers. Rupture of myocardial fibers was readily identified than in non-electrocution death. Sometimes, focal interstitial hemorrhage and polarization of endothelial cells were seen.</p><p><b>CONCLUSIONS</b>The electric marks on the skin, as confirmed by histologic examination, remain important sequelae of electrocution. The pathologic changes seen in myocardium provide additional clues to the diagnosis.</p>

The role of ouabain in cell death of vascular endothelial cells ECV304 and the changes of expression of Na+, K(+)-ATPase alpha1, beta1-subunit / 中国应用生理学杂志

<p><b>AIM</b>To study the effect of Na+, K(+)-ATPase inhibition by ouabain on growth and death of vascular endothelial cells ECV304 and involved mechanisms.</p><p><b>METHODS</b>Growth inhibition of ouabain on ECV304 cells was analyzed using MTT assay. The feature of cell death was studied by Hoechst 33342/PI staining, transmission electron microscopy and DNA agarose gel electrophoresis in ECV304 cells treated with ouabain. The mRNA expression of Na+, K(+)-ATPase alpha1, beta1-subunit was examined by reverse transcription PCR (RT-PCR).</p><p><b>RESULTS</b>Ouabain inhibited the growth of ECV304 cells in a dose and time-dependent manner. 10 micromol/L ouabain treated for 24 hours could stimulate the necrosis of ECV304 cells; When treated with 0.1 micromol/L ouabain for 24-48 hours, the cells showed obviously defluxion, the loss of cell-cell contacts, nuclear chromatin condensation, chromatin margination and DNA fragmentation. Na+, K(+)-ATPase alpha1-subunit mRNA expression was significantly up-regulated in ECV304 cells treated with ouabain while the beta1-subunit expression conversely showed a significant decrease.</p><p><b>CONCLUSION</b>Ouabain could up-regulate Na+, K(+)-ATPase alpha1-Subunit expression and reduce beta1-Subunit expression which mediated signal transduction and decreased cell-cell adhesions and induced ECV304 cells death.</p>

Effect of selenium supplementation on activity and mRNA expression of type 1 deiodinase in mice with excessive iodine intake / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the effect of selenium supplementation on the selenium status and selenoenzyme, especially the activity and mRNA expression of type 1 deiodinase (D1) in mice with excessive iodine (EI) intake and to explore the mechanism of selenium intervention on iodine-induced abnormities.</p><p><b>METHODS</b>Weanling female BALB/c mice were given tap water or 3 mg/L of iodine or supplemented with 0.5 mg/L or 1.0 mg/L of selenium in the presence of excessive iodine for 5 months. Selenium status, thyroid hormone level, hepatic and renal D1 activity and mRNA expression were examined.</p><p><b>RESULTS</b>Excessive iodine intake significantly decreased the selenium concentration in urine and liver, and the activity of glutathione peroxidase (GSH-Px) in liver. Meanwhile, serum total T4 (TT4) increased while serum total T3 (TT3) decreased. Hepatic D1 enzyme activity and mRNA expression were reduced by 33% and 86%, respectively. Renal D1 enzyme activity and mRNA were reduced by 30% and 55%, respectively. Selenium supplementation obviously increased selenium concentration, activity of GSH-Px and Dl as well as mRNA expression of D1. However, increasing the supplementation of Se from 0.5 to 1.0 mg/L did not further increase selenoenzyme activity and expression.</p><p><b>CONCLUSION</b>Relative selenium deficiency caused by excessive iodine plays an essential role in the mechanism of iodine-induced abnormalities. An appropriate dose of selenium supplementation exercises a beneficial intervention.</p>

Diagnostic value of high frequency color Doppler ultrasonography for epididymitis / 中华男科学杂志

<p><b>OBJECTIVES</b>To evaluate the diagnostic value of color Doppler ultrasonography for epididymitis.</p><p><b>METHODS</b>High frequency color Doppler ultrasonography was performed in 42 patients with epididymitis and 21 health volunteers.</p><p><b>RESULTS</b>Compared with control group, epididymis enlarged significantly all in head, body and tail in 92% of patients with epididymitis. Bilateral epididymitis occurred in 73% of patients. Color Doppler flow imaging (CDFI), in patient group, showed the markedly increased flow signals bright in bundle in epididymis. Mild hydroccele of testis and orchitis were companied in most patients(93%). Orchitis was often complicated in the patients with long history.</p><p><b>CONCLUSIONS</b>High frequency color Doppler ultrasonography can be a preferred exam method and has diagnostic value for epididymitis.</p>

Effects of long-term excessive Iodine intake on the maternal-fetal thyroid hormone metabolism in mice / 中华内分泌代谢杂志

Long-term excessive iodine intake resulted in an increased TT_4 level and a decreased TT_3 level in maternal serum,meanwhile,hepatic and renal type 1 deiodinase activity decreased dose-dependently.A significant reduction in type 2 deiodinase ( D2 ) activity of 12.5 d placenta was found in 3.0 mg/L or above groups.For 19.5 d uterus,D2 activity decreased and type 3 deiodinase activity increased.The results suggest that excessive iodine has an effect on the embryonic development by regulating maternal-fetal thyroid hormone metabolism.