Adult ; Computers, Handheld ; Data Collection ; Humans ; Male ; Otolaryngology ; User-Computer Interface

OBJECTIVETo study the resistant related molecules of human leukemia drug resistant K562 cells (K562/HHT) induced by homoharringtonine (HHT).

METHODSGene expression profiles on K562/HHT, K562 and K562/HHT/RU486 (K562/HHT reversed by RU486) cells were detected by DNA microarray. The bone marrow tyrosine kinase gene in chromosome X (BMX) which changed dynamically among the three cells was confirmed by RT-PCR and Western blot. Then, BMX was transfected into K562 and K562/HHT cells, and the changes of daunorubicin (DNR) concentrations in these two cells were observed for BMX overexpression.

RESULTSAs compared with K562, there were changes in 117 gene expressions in K562/HHT, 57 of which were up-regulated and 60 down-regulated. The mdrl gene was significantly up-regulated. When compared with K562/HHT, 50 significantly differently expressed genes were screened out in the K562/HHT/RU486 cells, of which up- and down-regulated genes were 13 and 37 respectively. These genes involved in drug resistance, cell signaling, cell differentiation, cell proliferation, transcription regulator, ion transport and so on. Four genes [NM-001721 (BMX), NM-031459 (SESN2), NM-033642 (FGF13) and AL-049309 (SFRS12)] expressed significantly differently in the two group cells, BMX gene expression was higher in K562/HHT, than in K562, but lower than in K562/HHT/RU486 as confirmed by RT-PCR and Western blot. After the plasmid pCI-neo-BMX was transfected into K562 and K562/HHT cells, DNR concentration was significantly lower (79.28 +/- 4.04, 29.84 +/- 2.67) than those before transfection (158.52 +/- 8.08, 58.58 +/- 6.53).

CONCLUSIONBMX is associated with multi-drug resistance of K562/HHT cell line.

Drug Resistance, Multiple ; drug effects ; genetics ; Drug Resistance, Neoplasm ; drug effects ; genetics ; Gene Expression Profiling ; Harringtonines ; pharmacology ; Humans ; K562 Cells ; Leukemia ; drug therapy ; genetics ; metabolism ; Protein-Tyrosine Kinases ; genetics ; metabolism

This study was aimed to screen human papillomavirus (HPV) types associated with esophageal squamous cell carcinoma of Kazakh in Xinjiang using the gene chip technique and study the clinical significance of this application. The DNAs were collected from esophageal squamous cell carcinoma tissues and healthy esophageal mucosa of Kazakh adults in Xinjiang, and amplified firstly using HPV MY09/11 and then using HPV G5+/6+ to screen positive HPV specimens. These positive specimens were further detected by the gene chip technique to screen highly pathogenic HPV types. After determination with nested PCR amplification with HPV MY09/11 and G5+/6+, the infection rate of HPV was 66.67% in the esophageal squamous cell carcinoma group and 12.12% in the healthy control group. By testing the positive HPV specimens from the esophageal squamous cell carcinoma group, the infection rate of HPV16 was 97.72% and the co-infection rate of HPV16 and HPV18 was 2.27%. HPV16 infection may be involved in the development of esophageal squamous cell carcinoma in Xinjiang Hazakh adults.

OBJECTIVE: To discuss the cause of disease, treatment and therapeutic effect in patients with rhegmatogenous retina detachment (RRD) combined with non-secondary glaucoma. METHODS: Clinical data of 28 patients with RRD combined with primary or congenital glaucoma were retrospectively analyzed. RESULTS: Twenty-five out of the 28 patients succeeded with one operation (89.3%). The intraocular pressure of post-operation:on the 1st day was 10 approximately 46 (28.1+/-6.5) mmHg, on the 7th day was (18.9+/-7.2) mmHg, and on the last re-examination day was (17.6+/-6.2) mmHg. Anti-glaucoma operation was performed in 10 patients after the retinal operation. Chroidal hemorrhage was found in 2 patients and 2 chroidal exudations were found after the retinal operation. CONCLUSION The proportion of primary open angle glaucoma is higher than that of primary angle closure glaucoma, and trauma or surgery before the retinal operation is an important cause in glaucoma patients with RRD. There is no obvious difference in the ratio of surgical success between non-secondary glaucoma with RRD and those RRD patients without glaucoma. Vitreotomy+ silicon oil injection or drainage of subretinal fluid+air injection+cryocoagulation+explants is recommended. Chroid is easily involved. It is important to control the intraocular pressure during and after the surgery. The final visual acuity is rather poor, which may be related to the glaucoma and intraocular pressure.
Adolescent ; Adult ; Aged ; Child ; Female ; Glaucoma ; complications ; surgery ; Humans ; Male ; Middle Aged ; Retinal Detachment ; complications ; surgery ; Retrospective Studies ; Visual Acuity ; Vitrectomy

OBJECTIVETo establish hepatitis C virus (HCV) infected cell model which is similar to the infection in vivo and can support HCV to replicate for a long time.

METHODSAfter infected with HCV-positive serum, Hep G2 cells were cultured for 60 days. Nested RT-PCR was used to detect plus and minus HCV RNA in cultured cells and supernatants.

RESULTSPlus HCV RNA was detected intermittently in Hep G2 cells during 2-30 days, minus HCV RNA was detected during 3-30 days after infection, the detection rate was similar to plus HCV RNA. Plus and minus HCV RNA can be still intermittently detected during 31-60 days after infection. However, the detection rate gradually declined. Plus HCV RNA was also found intermittently positive in the supernatant, and the detection rate was consistent to that in cells. Minus HCV RNA was not detected in the supernatant.

CONCLUSIONHep G2 cells were susceptible to HCV, and could support HCV to replicate for a relatively long time. Hep G2 is an ideal HCV infection cell model.

Cell Line, Tumor ; Hepacivirus ; genetics ; growth & development ; Humans ; RNA, Viral ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Virus Replication

OBJECTIVETo explore the effects of Di (2-ethylhexyl) phthalate (DEHP) on the testis and testicular gubernaculum of fetal KM mice in vivo and to investigate the mechanism of DEHP-induced cryptorchidism.

METHODSThirty healthy pregnant KM mice were randomly and equally divided into a blank control group, a corn oil control group and a DEHP group. The pregnant mice in the latter group were exposed to DEHP by gavage at the dose of 500 mg/kg body weight per day from gestation day 12 (GD12) through gestation day 19 (GD19). The effects of DEHP were observed on the number of fetuses per pregnancy, the ratio of male to female pups, the weight of the testis, the morphology and location of the testis and gubernaculum, the relative testis-bladder neck distance (TBD) and cranial suspensory ligament (CSL) residual. The expressions of the androgen receptor (AR), estrogen receptor (ER) and actin and proliferating cell nuclear antigen (PCNA) in the gubernaculum were detected by immunohistochemistry.

RESULTSDEHP reduced the testis weight and TBD, induced different degrees of testis maldescent, but produced no obvious effect on the body weight, the number of fetuses per pregnancy, the sex ratio and the testis gubernacular morphology. Under the light microscope, hypotrophy was seen in all the testis seminiferous tubules, spermatogenic cells and Sertoli cells, marked Leydig cell hyperplasia was noted, and the positive expression of AR in the gubernaculum was decreased in the DEHP group (P < 0.01).

CONCLUSIONDEHP could cause dysfunction of the testis gubernaculum via its anti-androgen effect, induce cryptorchidism, and cause dysplasia and dysfunction of Sertoli cells, Leydig cells and spermatogenic cells in fetal mice.

Animals ; Diethylhexyl Phthalate ; pharmacology ; Female ; Fetus ; drug effects ; Leydig Cells ; drug effects ; Male ; Mice ; Mice, Inbred Strains ; Pregnancy ; Sertoli Cells ; drug effects ; Testis ; cytology ; drug effects ; pathology

Adult ; Female ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; blood ; virology ; Humans ; Leukocytes, Mononuclear ; virology ; Male ; MicroRNAs ; metabolism ; Middle Aged ; Transcriptome ; Young Adult

OBJECTIVETo investigate the influence of human C-type natriuretic peptide (hCNP) on proliferation of vascular endothelial cells (HUVECs).

METHODSReconstructed pcDNA3.1 (+)/hCNP was transfected into HUVECs with polyethylenimine and its plasmid expression was examined with RT-PCR, immunohistochemistry and Western blot. MTT method was used to determine the effect of expressed protein on proliferation of HUVECs. pcDNA3.1 (+)/hCNP transfection was used for control.

RESULTSThe proliferation of HUVEC 48 h after pcDNA3.1 (+)/hCNP transfection was (0.301 +/- 0.096), which was obviously higher than that with pcDNA3.1 (+) transfection (0.164 +/- 0.012). Reconstructed pcDNA3.1 (+)/hCNP might be expressed in HUVECs effectively and its protein expression was capable of promoting HUVECs proliferation markedly.

CONCLUSIONThe successive expression of reconstructed pcDNA3.1 (+)/hCNP and the promoting activity of its expressed protein on HUVECs lay the foundation potential therapeutic value of C-type natriuretic peptide.

Cell Line ; Cell Proliferation ; Endothelial Cells ; cytology ; Humans ; Natriuretic Peptide, C-Type ; genetics ; Plasmids ; RNA, Messenger ; genetics ; Transfection

To explore the effects of vascular endothelial growth factor (VEGF) on the mechanisms of CML pathogenesis, the effect of VEGF on K562 cell apoptosis induced by As(2)O(3) was analyzed through morphologic observation, DNA fragmentation agarose gel electrophoresis and DNA ploidy flow cytometry analysis, and the effect of VEGF on the expression of bcl-X(L), Bax and caspase-3 in K562 cells was determined by Western blot, meanwhile the expression difference between bcl-X(L) and Bax mRNA in above conditions was detected by RT-PCR. The results showed that after VEGF added, the apoptosis of K562 cells reduced, however, there was no significant changes in cell cycle distribution (P > 0.05). At the same time, following the increasing of the concentration of VEGF, expression of mRNA and protein of bcl-X(L) was up-regulated and the expression of Bax protein was down-regulated in K562 cells, and the activation of pro-caspase-3 into caspase-3 was inhibited or reduced. It is concluded that VEGF may suppress the apoptosis of K562 cells through its influence on the bcl-X(L)/Bax expression ratio in K562 cells.
Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Blotting, Western ; Chlorides ; pharmacology ; Flow Cytometry ; Humans ; K562 Cells ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A ; pharmacology ; bcl-2-Associated X Protein ; biosynthesis ; genetics ; bcl-X Protein ; biosynthesis ; genetics

Dioscin has a wide range of biological effects and broad application prospects. However the studies concerning the toxicology and mechanism of dioscin is small. This article is to study the hepatotoxicity of dioscin and the effect of dioscin treatment on expression of aryl hydrocarbon receptor (AhR) mRNA and CYP1A mRNA and protein in HepG2 cells in vitro. Dioscin 0.5-32 µmol · L(-1) exposed to HepG2 cells for 12 h, cell viability was examined by CCK-8 assay and the release rate of lactate dehydrogenase (LDH) was to evaluate cell membrane damage. HepG2 cells morphologic changes were quantified by inverted Microscope, and the effect on production of reactive oxygen species (ROS) was detected by flow cytometry. The mRNA expression of CYP1A and AhR was evaluated by RT-RCR. The protein expression of CYP1A1 was detected by western blot. The cell viability was significantly inhibited after HepG2 cells were exposed to dioscin 0.5-32 µmol · L(-1). Compared with the control, the LDH release rate and ROS were significantly increased. The expression of CYPlA and AhR mRNA was increased. The expression of CYP1Al protein was increased after dioscin treatment, and resveratrol, an AhR antagonist, could downregulate the expression of CYP1A1. It follows that large doses dioscin has potential hepatotoxicity. The possible mechanism may be dioscin can active aryl hydrocarbon receptor (AhR) and induce the expression of CYP1A.
Cell Survival ; drug effects ; Chemical and Drug Induced Liver Injury ; etiology ; Cytochrome P-450 CYP1A1 ; genetics ; Diosgenin ; analogs & derivatives ; toxicity ; Hep G2 Cells ; Humans ; L-Lactate Dehydrogenase ; secretion ; RNA, Messenger ; analysis ; Reactive Oxygen Species ; metabolism ; Receptors, Aryl Hydrocarbon ; genetics