OBJECTIVETo detect the transcriptional level of a novel gene F10 associated with the pathogenesis of hydatidiform mole in human cell lines and screen the cell lines with low F10 expression to construct a stable eukaryotic expression system for F10 gene.

METHODSThe expression level of F10 mRNA was detected with fluorescent quantitative PCR in A549, 16HBE, Bel7402, HIC, HepG2, 293, PC and MGC cell lines. A549 cell line was transfected with plasmid pRc-CMV2-F10 via electroporation to allow stable F10 expression, and the positive cell clones were selected by G418. The insertion and expression of F10 gene in the A549 cells was analyzed using fluorescent quantitative PCR.

RESULTSF10 mRNA was expressed differentially in these cells lines, and the Bel7402 cells, PC and MGC cells showed the highest F10 mRNA expression, followed by HepG2 and HIC cells and further by 293 cells, and 16HBE and A594 cells had the lowest expression. After transfection, A594 cells showed genomic integration of F10 gene and high expression level of F10 mRNA.

CONCLUSIONThe pulmonary carcinoma cell line A549 with stable expression of F10 gene has been established, which may facilitate further study of the biological functions of F10 gene.

Cell Line, Tumor ; Eukaryotic Cells ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Genes, Neoplasm ; genetics ; Humans ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Trophoblastic Neoplasms ; genetics ; pathology ; Uterine Neoplasms ; genetics ; pathology

OBJECTIVETo study the function of F10 gene, a novel hydaditiform mole-related gene.

METHODSA549 cell line was transfected with the F10 gene of forward or reverse sequence or with the empty vector, respectively. The cellular mRNA was extracted after 24 h of transfection to screen for the differentially expressed genes among the 3 transfected and the control cells using differential display-polymerase chain reaction (ddPCR).

RESULTSThe bands representing differentially expressed genes were amplified from the cells, and the products were linked to T-Vector for sequence analysis. Several genes were screened by Blasting and their expressions were confirmed by fluorescent quantitative PCR.

CONCLUSIONF10 gene is functionally related to cell proliferation and apoptosis.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Epithelial Cells ; metabolism ; Female ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Hydatidiform Mole ; genetics ; Hydatidiform Mole, Invasive ; genetics ; Lung Neoplasms ; genetics ; pathology ; Oncogenes ; genetics ; Pregnancy ; Transfection ; Uterine Neoplasms ; genetics

Larp4b is a member of the LARP family, which can interact with RNA and generally stimulate the translation of mRNA. Abnormal expression of Larp4b can be found in leukemia patients in our previous study. This study was purposed to detect the relative expression of Larp4b mRNA in different subpopulations of mouse hematopoietic cells, to construct lentivirus vector containing shLarp4b targeting mouse gene Larp4b and to explore its effects on mouse Lin(-) cells infected with shLarp4b by lentivirus. SF-LV-shLarP4b-EGFP and control vectors were constructed and two-plasmid lentivirus packing system was used to transfect 293T cells. After 48 h and 72 h, lentivirus SF-LV-shLarp4b-EGFP was harvested and was used to infect Lin(-) cells. After 48 h, EGFP(+) cells was sorted by flow cytometry (FCM). Meanwhile, semi-quantitative real time-PCR, AnnexinV-PE/7-AAD staining, PI staining and colony forming cell assay (CFC) were performed to determine the expression of Larp4b and its effect on the proliferation of hematopoietic progenitor cells. The results showed that Larp4b was highly expressed in myeloid cells. SF-LV-shLarp4b-EGFP was successfully constructed according to the restriction endonuclease digestion assay. RT-PCR confirmed that Larp4b was efficiently knockdown in mouse Lin(-) cells. The low expression of Larp4b did not affect the colony forming number, the apoptosis and cell cycle of Lin(-) cells. It is concluded that knockdown of Larp4b in mouse Lin(-) cells do not contribute to the colony forming ability and the growth of Lin(-) cells in vitro. This useful knockdown system will be used to study in vivo Larp4b in future.
Animals ; Autoantigens ; metabolism ; Cells, Cultured ; Flow Cytometry ; Gene Knockdown Techniques ; Genetic Vectors ; Hematopoietic Stem Cells ; cytology ; Humans ; Lentivirus ; genetics ; Mice ; Plasmids ; Ribonucleoproteins ; metabolism ; Transfection

OBJECTIVETo evaluate the safety and effectiveness of endourological techniques in the treatment of benign prostate hyperplasia (BPH) in aged high-risk patients.

METHODSWe used endourological techniques in the treatment of 283 BPH patients aged over 70 years and complicated with hydronephrosis, renal failure, heart failure, cerebral infarction, respiratory dysfunction, anemia, diabetes, bladder tumor, or prostate weight over 80 g, TURP (transurethral resection of the prostate) for 112 cases and PKRP (transurethral plasmakinetic resection of the prostate) for the other 171. All the patients were followed up for 1-30 months.

RESULTSIn the TURP group, the scores on IPSS and QOL were decreased from 27.5 +/- 2.8, 5.5 +/- 1.0 to 5.8 +/- 1.2, 1.0 +/- 0.5, and the residual urine volume (RUV) from (75.0 +/- 20.0) ml to (8.0 +/- 3.0) ml, but the maximal flow rate (Qmax) increased from (6.5 +/- 2.0) ml/s to (18.5 +/- 1.5) ml/s (P < 0.05), while in the PKRP group, the scores on IPSS and QOL were decreased from 28.2 +/- 2.2, 5.5 +/- 1.0 to 5.4 +/- 1.6, 1.0 +/- 0.5, and RUV from (80.0 +/- 20.0) ml to (7.0 +/- 3.0) ml, and Qmax increased from (6.8 +/- 2.1) ml/s to (20.0 +/- 1.5) ml/s (P < 0.05). There were no statistically significant differences in IPSS, QOL, Qmax and RUV after treatment between the two groups (P > 0.05), but significantly less complications were found in the PKRP than in the TURP group (P < 0.05).

CONCLUSIONEndourological treatment, especially PKRP, with comprehensive perioperative preparations, unerring operative skills, well-controlled operation time, and intensive postoperative monitoring and nursing, has the advantages of high safety, less bleeding, fewer complications and definite effectiveness for aged high-risk BPH patients.

Aged ; Aged, 80 and over ; Humans ; Male ; Prostatic Hyperplasia ; surgery ; Quality of Life ; Transurethral Resection of Prostate ; methods ; Treatment Outcome

The rhizome of Alpinia officinarum is a widely used Chinese herbal medicine. The essential oil in A. officinarum rhizome is mainly composed of 1, 8-cineole and other monoterpenes, as the major bioactive ingredients. In plants, monoterpenes are synthesized through the methylerythritol phosphate (MEP) pathway in the plastids, and 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) is an enzyme catalyzing a committed step of the MEP pathway. In the present study, the full-length cDNA encoding DXR was cloned from the rhizome of A. officinarum, using homology-based RT-PCR and rapid amplification of cDNA ends (RACE) techniques. The new cDNA was designated as AoDXR and submitted to GenBank to be assigned with an accession number HQ874658. The full-length cDNA of AoDXR was 1 670 bp containing a 1 419 bp open reading frame encoding a polypeptide of 472 amino acids with a calculated molecular mass of 51.48 kDa and an isoelectric point of 6.15. Bioinformatic analyses revealed that AoDXR showed extensive homology with DXRs from other plant species and contained a conserved plastids transit peptide, a Pro-rich region and two highly conserved NADPH-binding motifs in its N-terminal region characterized by all plant DXRs. The phylogenetic analysis revealed that AoDXR belonged to angiosperm DXRs. The structural modeling of AoDXR showed that AoDXR had the typical V-shaped structure of DXR proteins. The tissue expression pattern analysis indicated that AoDXR expressed strongly in leaves, weak in rhizomes of A. officinarum. Exogenous methyl jasmonate (MeJA) could enhance the expression of AoDXR and the production of 1, 8-cineole in A. officinarum rhizomes. The cloning and characterization of AoDXR will be helpful to reveal the molecular regulation mechanism of monoterpene biosynthesis in A. officinarum and provides a candidate gene for metabolic engineering in improving the medicinal quality of A. officinarum rhizome.
Aldose-Ketose Isomerases ; genetics ; Alpinia ; chemistry ; enzymology ; genetics ; Amino Acid Sequence ; DNA, Complementary ; genetics ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Plant ; Molecular Sequence Data ; Monoterpenes ; metabolism ; Phylogeny

OBJECTIVETo evaluate the static and dynamic contrast sensitivity changes in myopic patients before and after laser in situ keratomileusis (LASIK).

METHODSSeventy-three eyes in 37 patients with myopia (with or without astigmatism) who received LASIK were tested for static and dynamic contrast sensitivities using the METRO VISION MON ELEC I system at 0.7, 1.4, 2.7, 5.5, 11, and 22 cpd and cps prior to LASIK, and at one-, three-, and six-month intervals after LASIK.

RESULTSAll eyes gained naked visual acuity of more than 0.5 after LASIK. The contrast sensitivity was depressed at all frequencies 1 month after LASIK, as compared to one week prior to LASIK. The depression at 2.7, 5.5, 11 (P < 0.01) and 22 cpd (P < 0.05) was statistically significant for static contrast sensitivity, and also at 5.5 (P < 0.01) and 11 cps (P < 0.05) for dynamic contrast sensitivity. Myopic eyes between 6.25 D and 14.0 D, and astigmatic eyes 2 DC and more, suffered more static and dynamic contrast sensitivity depression than the myopic eyes between 1.25 D and 6.00 D and astigmatic eyes less than 2 DC. Contrast sensitivities were improved and exceeded preoperative levels 3 months after LASIK, and improved even more 6 months after LASIK. All sequences were statistically significant for static contrast sensitivity (P < 0.01), while only 2.7, 5.5, and 11 cps were statistically significant for dynamic contrast sensitivity (P < 0.01). The astigmatic eyes 2 DC and more showed less improvement, even below the preoperative level at 1.4 cps of dynamic contrast sensitivity.

CONCLUSIONSWhile temporary depression of contrast sensitivity for myopic eyes after LASIK was seen, contrast sensitivity soon returned to exceed preoperative levels at 3 months after LASIK, while improving even more 6 months after LASIK.

Adolescent ; Adult ; Astigmatism ; surgery ; Contrast Sensitivity ; Cornea ; surgery ; Female ; Humans ; Keratomileusis, Laser In Situ ; Male ; Myopia ; physiopathology ; surgery ; Visual Acuity

To evaluate the differences of Ophiopogonjaponicus from different cultivations, the metabolomics based method was conducted to compare the effects of Hangmaidong and Chuanmaidong (Chinese name) on plasma endogenous metabolites of normal rats. Data were collected by ultra performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-Q-TOF/MS), and were analyzed by multivariate statistical method, such as Principal Component Analysis and Orthogonal Signal Correction Partial Least Square Discriminant Analysis. Results revealed that the plasma metabolites profiling of low and middle dose group of Chuanmaidong were similar to the control group, but different from the high dose group obviously. Meanwhile the high, middle and low dose groups of Hangmaidong were different from control group notably, and the difference is dose dependent. Lysophosphatidylcholines, the possible endogenous metabolites which contribute to the classification most significantly, are closely related to cardiovascular system diseases. Compared with the group of Chuanmaidong, Hangmaidong has greater impact on the plasma metabolic profiling of normal rats. Hangmaidong and Chuanmaidong showed significant differences pharmacodynamically.
Animals ; Biomarkers ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; pharmacology ; Least-Squares Analysis ; Mass Spectrometry ; Metabolomics ; Principal Component Analysis ; Rats

OBJECTIVETo investigate the clinical features of mild encephalitis/encephalopathy with a reversible splenial lesion (MERS) in children.

METHODSThe clinical data of 8 children with MERS were retrospectively analyzed.

RESULTSThe mean age of onset was 5 years and 2 months (range 10 months to 12 years). The major clinical features included a history of prodromal infection, and among these children, 5 had pyrexia and 4 had vomiting. Of all the children, 6 were manifested as convulsion and 3 each were manifested as disturbance of consciousness and paroxysmal paropsia. Cranial diffusion-weighted magnetic resonance imaging (MRI) showed high signals in the splenium of the corpus callosum. Among these children, one child had symmetric and multiple long T1 and long T2 signals in the bilateral centrum semiovale and part of the temporal white matter. MRI reexamination performed after 5-30 days showed the disappearance of abnormal signals in all the children. The children were followed up for 3 months to 2 years, and no child experienced abnormal neurodevelopment.

CONCLUSIONSThe development of MERS in children is closely associated with infection. MERS is characterized by high signals in the splenium of the corpus callosum on cranial diffusion-weighted MRI. Most children have good prognosis.

Brain Diseases ; pathology ; Child ; Child, Preschool ; Corpus Callosum ; pathology ; Encephalitis ; pathology ; Female ; Humans ; Infant ; Magnetic Resonance Imaging ; methods ; Male ; Retrospective Studies

OBJECTIVETo understand the sexual physiology and psychology of male college students and to provide schools, families and the society with reference for the sexual physiological and psychological education among college students as well as for the diagnosis and treatment of their sexual psychological disorders in Jiangsu.

METHODSAn investigation was conducted by using a questionnaire on sexual physiology and psychology among randomly selected 3786 male college students from 18 universities in Jiangsu.

RESULTSAs regards sexual education, 5.49% of the subjects were satisfied with their schools, 78.18% wanted it to be strengthened and 68.36% obtained their sexual knowledge from the internet. Concerning sexual physiology, 68.78% experienced their first spermatorrhea at the age of 12-15. As for sexual psychology, 85.79% loved a certain female inwardly, and 70.99% experienced love affairs. With regard to sexual activity, 25.54% had sexual experience.

CONCLUSIONCollege students nowadays are relatively open in sexual ideology, immature in sexual psychology and lacking in sexual knowledge, while schools are inefficient in sexual education. Their sexual health calls for joint attention from schools, families and the society, particularly schools, which need to establish special offices for research and education on sexual health.

China ; Health Knowledge, Attitudes, Practice ; Humans ; Male ; Sexual Behavior ; physiology ; psychology ; Students ; psychology ; Universities ; Young Adult

OBJECTIVETo evaluate the efficacy of the monotherapy of 15 agents in treating essential hypertension.

METHODSAfter 2-week wash-out, a total of 370 patients with seated diastolic blood pressure 95-114 mmHg and seated systolic blood pressure < 180 mmHg were randomized to different therapeutic groups. 24-hour ambulatory blood pressure monitoring was performed before medication and at the end of 8 weeks.

RESULTAll the agents significantly reduced the 24 hour mean blood pressures after treatment except doxazosin, terazosin, and torasemide.

CONCLUSIONThe result suggested that the angiotensin-converting enzyme inhibitors, angiotensin II receptor blockers, beta-blockers and long-acting calcium antagonists were effective in treating essential hypertension, while the low-dose doxazosin, terazosin and torasemide can be used for combination therapy but not for monotherapy.

Adrenergic beta-Antagonists ; therapeutic use ; Angiotensin Receptor Antagonists ; Angiotensin-Converting Enzyme Inhibitors ; therapeutic use ; Antihypertensive Agents ; classification ; therapeutic use ; Blood Pressure Monitoring, Ambulatory ; Calcium Channel Blockers ; therapeutic use ; Doxazosin ; therapeutic use ; Drug Therapy, Combination ; Humans ; Hypertension ; drug therapy ; Prazosin ; analogs & derivatives ; therapeutic use ; Sulfonamides ; therapeutic use ; Treatment Outcome