OBJECTIVE:To establish a method for simultaneous determination of 4 psoralen compounds in Buwu tincture. METHODS:HPLC was performed on the column of Dikma Diamonsil C18 with mobile phase of acetonitrile-0.2% Acetic acid by gradient elution at flow rate of 1 ml/min,detection wavelength was 246 nm,column temperature was 30 ℃,and the injection vol-ume was 15 μl. RESULTS:The linear range was 13.00-208 μg/ml for angelicin,26.00-416 μg/ml for bavachin,24.50-392 μg/ml for psoralidin and 37.88-606 μg/ml for isobavachalcone,respectively(r≥0.999 6);RSDs of precision,stability and reproducibility tests were less than 2.00%;recoveries were 95.22%-97.23%(RSD=0.87%,n=6),100.24%-104.64%(RSD=1.62%,n=6), 102.28%-104.39%(RSD=1.47%,n=6)and 97.68%-100.17%(RSD=0.97%,n=6),respectively. CONCLUSIONS:The method is simple and reproducible,and can be used for the quality control of Buwu tincture.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Hep G2 Cells ; Humans ; Interleukin-6 ; metabolism ; Liver Neoplasms ; metabolism ; pathology

Background Intravitreal injection with triamcinolone acetonide (TA) may cause complications,including increase of intraoculapressure (IOP),cataracand endophthalmitis.Ketorolatromethamine (Ketorolac) inew,lesadverse reactionof non-steroidal anti-inflammatory drug.The action mechanism of Ketorolaisimilato TA.Therefore,Ketorolamay be completely opartly replace Tin the treatmenof retinal edema.Objective The purpose of thistudy wato investigate the effectof Tand Ketorolaon the expressionof aquaporin-4 (AQP4) and vasculaendothelial growth facto(VEGF) in hypoxiretinal Müllecellin vitro and to explore the mechanism of treating retinal edemwith Tand Ketorolac.MethodThe propose of research and use of the animalwere approved by Animal ExperimenResearch Review Committee of Hubei University of Medicine.Twenty eyeof New Zealand albino rabbitwere extracted and the retinal tissue waisolated.The Müllecellwere cultured and passaged using the enzymatidigestion method and Müllecellwere identified using glial fibrillary acidiprotein (GFAP),vimentin and α-smooth muscle actin (α-SMA) by immunofluorescence staining.The hypoxicell modelwere established by culturing the cellin DMEM with 500 μmol/L CoCl2 fo0,6,12,24 hours.The cellof hypoxifo24 hourwere divided into normal control group,hypoxicontrol group,hypoxia+50,100,200 mg/L To50,100,200 mg/L Ketorolagroups.Corresponding drugwere added into the medium in the differengroups.The expressionof AQP4 mRNand VEGF mRNin Müllecellwere detected by semi-quantitative reverse transcription PC(RT-PCR).ResultThe cellgrew well and reached 80％ confluence with the irregulashape and ovoid nuclei 14-15 dayaftecultured.More than 95％ primary cellshowed positive reaction to GFAP,vimentin and α-SMA.The expressing levelof AQP4 mRNand VEGF mRNin Müllecell(values) were significantly differenin varioutime point(AQP4 mRNA:F=18.70,P＜0.01 ; VEGF mRNA:F =53.20,P＜0.01),and those of 6,12 and 24 houraftecultured with CoCl2were increased than those withouCoCl2 (P＜0.05).The expressing levelof AQP4 mRNand VEGF mRNin Müllecell(values) were significandifferenamong the normal control group,hypoxicontrol group,hypoxia+50,100,200 mg/L ToKetorolagroup(AQP4 mRNA:F =27.98,P ＜ 0.01 ; VEGF mRNA:F =10.03,P ＜0.01).Compared with the hypoxicontrol group,the expressing levelof AQP4 mRNand VEGF mRNin the Müllecellwere declined in the hypoxia+ 100,200 mg/L Tgroup and the hypoxia+100,200 mg/L Ketorolagroup (P＜0.05).The expressing levelof AQP4 mRNand VEGF mRNwere found statistically insignificandifference between hypoxia+ 100 mg/L Tgroup and hypoxia+ 100 mg/L Ketorolagroup,obetween hypoxia+ 200 mg/L Tgroup and hypoxi+200 mg/L Ketorolagroup (P＞ 0.05).ConclusionTand Ketorolacan downregulate the expressionof AQP4 and VEGF in Müllecellundehypoxiconditions,inferring thathey have similamechanism in the impacon AQP4 function in retinal edematoueye.

OBJECTIVETo determine the effective dose of hepatitis B immunoglobulin (HBIG) for clearing maternally-transmitted hepatitis B virus (HBV) from a newborn.

METHODSFull-term neonates born to HBV-infected mothers were tested for hepatitis B surface antigen (HBsAg) and HBV DNA in venous blood, Individuals with positive results within two hours after birth were selected for study, and divided among two treatment groups: research group receiving HBIG continually adjusted to quantitative levels of neonatal HBsAg and HBV DNA levels; control group receiving standard HBIG 200IU dose. All neonates were also treated with 10 micrograms of recombinant vaccine. The decreases in HBsAg and HBV DNA over 12 months were comparatively analyzed between the two treatment groups.

RESULTSThe two treatment groups (HBIG adjusted vs. standard) were statistically similar in Apgar score (9.38+/-0.49 vs. 9.37+/-0.48), neonate body weight (3458.67+/-374.93 vs. 3558.61+/-322.85 g), maternal age (26.33+/-3.63 vs. 25.33+/-3.03), and initial HBsAg and HBV DNA levels (rank sum test Z = 1.381, and Z = 0.700, respectively) (all, P more than 0.05). Successful clearance of HBV infection within 12 months was achieved in significantly more neonates in the HBIG adjusted therapy group than in the standard therapy group (82.8% vs. 57.4%; x2 = 9.696, P less than 0.05).

CONCLUSIONAdjusting the neonatal HBIG dose according to HBsAg and HBV DNA levels can improve the success rate of clearing maternally-transmitted HBV.

DNA, Viral ; Dose-Response Relationship, Immunologic ; Female ; Hepatitis B ; prevention & control ; therapy ; Hepatitis B Surface Antigens ; blood ; immunology ; Hepatitis B virus ; genetics ; immunology ; Humans ; Immunoglobulins ; administration & dosage ; therapeutic use ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; prevention & control

OBJECTIVETo observe effects of electroacupuncture (EA) on gastric blood perfusion and search for a study method reflecting changes of blood perfusion.

METHODSOn the basis of establishing a new gastric ischemia-reperfusion model which was made by ligature of gastric right artery, EA was given at "Zusanli" (ST 36) and the gastric blood perfusion images were displayed by a laser Doppler blood perfusion imager (LDPI), and effects of EA on gastric blood perfusion were analyzed.

RESULTS(1) The image of the gastric ischemia-reperfusion is clear. The ischemic degree of the stomach in the EA group was lower than the control group within 30 min of ischemia, and recovery of blood flow after perfusion in the EA group was better than the control group; (2) at 25 min of ligature, there was a significant difference in changes of blood perfusion between the EA group [(-0.50 +/- 0.18) PU] and the control group [(-0.90 +/- 0.16) PU], P < 0.05.

CONCLUSIONEA can improve and promote gastric blood supply in the gastric ischemia-reperfusion model, which can be used for acupuncture and moxibustion experimental study.

Animals ; Electroacupuncture ; Female ; Laser-Doppler Flowmetry ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; physiopathology ; therapy ; Stomach ; blood supply

OBJECTIVETo investigate the expression of Oct4, Sox2, Nanog, SMO, beta-Catenin and Wnt5b mRNA in four hepatocellular carcinoma cell lines of SMMC-7721, Bel-7402, HepG2, MHCC-97 and normal hepatocellular cell line of L02, and to compare the response of these cell lines to all-trans retinoic acid.

METHODSRT-PCR was used to detect expression of Oct4, Sox2, Nanog, SMO, beta-Catenin and Wnt5b mRNA in four hepatocellular carcinoma cell lines and normal hepatocellular cell line. Real time-PCR was used to quantify the expression of the genes.

RESULTSThere are different levels of expression of the stem cell-related gene in hepatocellular carcinoma cell lines and control cell line (P less than 0.05). There are significant differences in HepG2 and L-02 for the response to all-trans retinoic acid (P less than 0.05).

CONCLUSIONSThe stem cell-related genes are differentially expressed in different hepatocellular carcinoma cell lines.

Carcinoma, Hepatocellular ; metabolism ; pathology ; DNA Methylation ; Gene Expression Regulation, Neoplastic ; Hep G2 Cells ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Nanog Homeobox Protein ; Octamer Transcription Factor-3 ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Receptors, G-Protein-Coupled ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; SOXB1 Transcription Factors ; genetics ; metabolism ; Signal Transduction ; Smoothened Receptor ; Stem Cells ; drug effects ; metabolism ; Tretinoin ; pharmacology ; Wnt Proteins ; genetics ; metabolism ; beta Catenin ; genetics ; metabolism

OBJECTIVETo evaluate the impact of specific siRNA on survivin gene in transfected leukemia cells.

METHODThe small interfering RNA (siRNA) targeted survivin mRNA was synthesized in vitro and was transfected into K562 cell by Hiperfect into human leukemia cell line K562, which has high survivin expression level. The level of survivin mRNA expression was determined by quantitative reverse transcription polymerase chain reaction (RT-PCR) with SYBR GREEN I. The apoptosis index of cytotrophoblasts were determined and analyzed by FCM (Annexin V-FITC/PI staining methods). The cell proliferation was examined by MTT at 48 h and 72 h after transfection.

RESULTThe level of mRNA expression was significantly inhibited by the siRNA 48 h and 72 h after transfection, the suppression rate of survivin mRNA separately reached 85.21%, 94.35% mensurated by quantitative RT-PCR with SYBR GREEN I, cell proliferation was inhibited significantly by 45.02% and 50.88%, respectively, the apoptotic rate detected by Annexin V-FITC assay reached 12.28%and 21.55%, respectively.

CONCLUSIONThe chemosynthesized siRNA targeting survivin could significantly down-regulate survivin mRNA. Survivin siRNA was able to inhibit the proliferation of leukemia cell line K562. Survivin may become a new target for leukemia gene therapy.

Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Gene Silencing ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; K562 Cells ; RNA, Small Interfering ; pharmacology ; Transfection

OBJECTIVETo evaluate a finite element method (FEM) for analysis of the cranial-facial morphology.

METHODSThe two-dimensional finite element analysis system was established and used to analysis the lateral side morphology of the soft tissue by the change of each finite unit of the soft tissue in a X-ray cranial-facial lateral cepholometrics film.

RESULTSThe finite element analysis system was showing very well in the figures and data made by the system.

CONCLUSIONFinite element analysis system may be a good supplement of the traditional X-ray cephalometrics to the soft tissue of orthognatics.

Cephalometry ; methods ; Face ; anatomy & histology ; Finite Element Analysis ; Humans ; Orthognathic Surgery

OBJECTIVETo identify the risk factors for otitis media with effusion (OME) in some kindergarten children in Wuhan City of China and analyze the results with reference to the review of the literature.

METHODSThe study subjects were 3 to 6 years old children drawn from a school screening program for OME in Wuhan. All subjects were assessed with routine otorhinolaryngologic examination, otoscopic examination and tympanometry. During the test, parents were interviewed to provide information with regard to the children's birth history, neonatal history, feeding history, family smoke history, otological history, rhinorrhea, sneeze, nasal obstruction, snoring, tonsillitis episodes history, et al. These data formed the basis in the estimation of potential risk factors for OME.

RESULTSIn the univariate analysis of 144 cases and 288 controls, significantly elevated odds ratios (OR) for OME were detected on the symptoms of nasal obstruction (OR = 2.60, P = 0.002), rhinorrhoea (OR = 1.442, P = 0.003), high hard palate (OR = 4.411, P < 0.0001), and previous history of acute otitis media (OR = 1.77, P = 0.025). However, four factors were found to be significant in the multivariate logistic regression model, including feeding history (OR = 0.746, P = 0.047), nasal obstruction (OR = 2.56, P = 0.003) and previous acute otitis media episodes (OR = 1.735, P = 0.032).

CONCLUSIONSPrevious acute otitis media episodes and nasal obstruction were risk factors, which was accordant with literature. In addition, breastfeeding was a protective factor for OME, but acute tonsillitis was not a factor for OME. A child who had previous acute otitis media episode and often had nasal obstruction is suggested to have otorhinolaryngologic regulatory examination.

Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Male ; Otitis Media with Effusion ; epidemiology ; etiology ; Risk Factors ; Surveys and Questionnaires

OBJECTIVETo investigate the serum levels of sCD44v6 and sE-cadherin (sE-cad) in patients with esophageal squamous cell carcinoma.

METHODSThe serum samples were collected from 65 cases of esophageal squamous cell carcinoma, 32 cases of erosive esophagitis and 35 healthy subjects. Serum sCD44v6 and sE-cad levels were measured by enzyme linked immunosorbent assay (ELISA).

RESULTSThe mean levels of serum sCD44v6 and sE-cad in esophageal squamous cell carcinoma patients were significantly higher than those of erosive esophagitis patients and normal controls (both P<0.05). There was no significant difference in serum sCD44v6 and sE-cad levels between erosive esophagitis patients normal controls (P=0.566 and P=0.708, respectively). Serum sCD44v6 and sE-cad levels of esophageal cancer patients were not correlated with their clinicopathological features. Serum sCD44v6 level is not correlated with sE-cad level in squamous cell carcinoma patients(P=0.651).

CONCLUSIONSerum sCD44v6 and sE-cad might be a potential marker for screening of esophageal squamous cell carcinoma.

Adult ; Aged ; Aged, 80 and over ; Cadherins ; blood ; Carcinoma, Squamous Cell ; blood ; pathology ; Case-Control Studies ; Esophageal Neoplasms ; blood ; pathology ; Female ; Humans ; Hyaluronan Receptors ; blood ; Male ; Middle Aged