Objective: To construct a recombinant adenovirus vector carrying human cytomegalovirus (hCMV) UL144 gene and to explore the biological characteristics of UL144 gene-modified DCs. Methods: The UL144 gene was amplified from hCMV DNA, which was extracted from hCMV-DNA positive serum. The recombinant adenovirus vector carrying hCMV UL144 gene was constructed with AdEasy system and then transfected into HEK293 cells to create recombinant adenovirus Ad-UL144. The expression of inserted gene was identified by RT-PCR. The recombinant adenovirus was then transfected into mice myeloid dendritic cells. The surface proteins of dendritic cells were analyzed by FACS, and cytokines in supernatant were detected by ELISA. T cell proliferation stimulated by gene-modified DC was examined by 3H-TdR uptake assay. Results: The UL144 gene was successfully cloned into the pAdEasy-1 plasmid. The recombinant adenovirus Ad-UL144 was packed in HEK293 cells, with a viral titer of 3×10 10 pfu/ml. DCs infected with AdCMV-UL144 had markedly decreased surface expression of CD80, CD86 and I-Ad (P < 0.01). The contents of TNF-α, IL-6 and IL-1β were significantly decreased in the supernatant of AdCMV-UL144 modified DCs (P < 0.05). T cell proliferation ability induced by gene-modified DC was obviously lower than in the DC control group (P < 0.01). Conclusion: UL144-modified DCs can maintain a relative immature status, and have reduced stimulating activity upon the proliferation and activation of T cells in vitro.

Objective To study the MRI findings and di agnostic value as well as pathological base of the knee iniury.Methods MRI features of knee injuries in 31 cases were r etrospectively analysed compared with the findings of operations and clinical ma nifestations.Results MRI findings were :(1)para-knee articular soft tissue swelling in 23 cases(74%);(2)MRI detected ligaments injury in 18 cas es (58%),dislocation of articular in 2 cases(6%);(3)bone contusions in 4 c ases(13%);occult fractures in 1 case(3%),disruption of articular cartilage 1 case(3%),fractures in 12 cases(39%);(4)meniscal injury in 15 cases(48% ),meniscal cyst in 2 cases(6%);(5)quadriceps femoris and kneecap tendon inj ury in 1 case(3%)respectively;(6)joint hydrocele in 29 cases(93%).Conclusion MRI can find a variety of pathological changes of knee injury and plays an important role in clinical treatment and predicting prognosis.

Background Argon laser peripheral iridoplasty (ALPI) is proved to be effective in lowering intraocular pressure (IOP) of patients with mild acute primary angle closure (APAC).It is unclear whether this laser treatment is equally efficient in managing patients with severe APAC.This study aimed to evaluate the IOP-lowering efficacy of ALPI and laser peripheral iridotomy (LPI) on patients with refractory APAC,who have previously responded poorly to intensive medical therapy.Methods Thirty-six patients (8 men and 28 women) were identified as medically refractory APAC,who still had ocular pain,red eye,hazy cornea,closed anterior chamber (AC) angle,and IOP of not less than 21 mmHg after two days or more of anti-glaucoma medication.All enrolled patients underwent ophthalmologic examinations including measurement of visual acuity (VA),best corrected VA (BCVA),IOP,biomicroscopy,and gonioscopy followed by ALPI immediately in the APAC eye and LPI in both eyes.Results All patients were affected unilaterally,with average age of (54.6±11.7) (range,37.0-75.0) years old.The mean IOP value of the affected eyes dropped from (31.6±7.7) (range,21.0-39.0) mmHg at enrollment to (18.4±8.7) (range,10.0-27.0) mmHg 2 hours after ALPI.At follow-up day 7,the mean IOP value maintained at (14.8±4.2) (range,9.0-21.0)mmHg,which was significantly different (P=0.000) compared with baseline.The average decrease of IOP in the APAC eyes was (16.8±7.4) (range,12.0-21.0) mmHg.At follow-up three years later,the mean IOP of the APAC eyes stabilized at (16.3±3.2) (range,9.0-20.0) mmHg with at least 180° of AC angle opened.Conclusion ALPI and LPI lower the IOP of medically refractory cases of APAC though they have responded poorly to anti-glaucoma medication.

Objective To explore the relationship between X - linked spondyloepiphyseal dysplasia tarda (SEDL) gene escaping X chromosome inactivation( XCI) and SEDL phenotype. Methods RT - PCR was performed on total RNA which was isolated from blood samples of patients, female carriers and controls. Patients and female carriers were selected from the pedigree with SEDL caused by the mutation (IVS2 - 2A→C) of the gene. cDNA was analyzed by polyacrylamide gelelectrophoresis(PAGE). Results PAGE data indicateed that female carriers expressed both normal and mutant SEDL mRNA,meaning the SEDL gene escaping XCI. Family investigation showed carrier females in the SEDL pedigree presented no symptoms. Conclusions The SEDL gene escaping X chromosome in-activation is firstly identified from human body. This may explain that carrier females present no symptoms.

Objective To investigate the risk factors and prognosis on the post-resuscitation multiple organ dysfunction syndrome(MODS) after successful cardiopulmonary resuscitation(CPR).Methods Clinical data of 53 patients who were suffered with cardiac arrest(CA) and undergone successful CPR with return of spontaneous circulation(ROSC) were analyzed. Results There were 48 patients accompanied with MODS(90.6%),and among them,35 patients died in hospital(66.0%).All the 10 patients with CPR interval ≥6min were accompanied with MODS and they died in hospital.There were 43 patients who underwent CA immediately and of them,38 patients were accompanied with MODS.The incidence and mortality of MODS in the patients with CA-ROSC interval 0.05).Conclusion The risk factors such as ROSC interval ≥6 min,AC-ROSC interval ≥10min and the SIRS after ROSC are significantly associated with the incidence of MODS.The organic function of the patients should be evaluated promptly.

Allele-specific polymerase chain reaction (AS-PCR) is a technique based on allele-specific primers, which can be used to analyze single nucleotide polymorphism (SNP) effectively including the transition, transversion and insertion/deletion polymorphism and has been exploited in the study of diseases research, molecular diagnosis, and forensic biological evidence. The article systematically reviews the principle, the detection methods, improvement of AS-PCR, and its research updates in the fields of autosome, Y chromosome and mitochondrial SNP, as well as its application in forensic science.
Alleles ; DNA Primers ; Forensic Sciences ; Humans ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Single Nucleotide

AIMTo study the protein binding of glimepiride.

METHODSAn HPLC-FA method is performed by using Pinkerton GFF II-S5-80 internal-surface reversed-phase silica support (150 mm x 4.6 mm ID, 5 microm) at pH 7.4 in a 67 mmol x L(-1) isotonic sodium phosphate buffer at 37 degree C. Other conditions included flow rate of 0.2 mL x min(-1), UV detection at wavelength 230 nm and injection volume 900 microL.

RESULTSNonlinear regression parameter estimation was used for the association constant measurement of glimepiride to both primary and secondary sites, which were 5.1 (micromol x L(-1)-1 and 1 for K1 and n1, and 0.017 (micromol x L(-1))-1 and 7 for K2 and n2, respectively.

CONCLUSIONThe method is shown to be suitable for investigation of protein binding of glimepiride.

Chromatography, High Pressure Liquid ; methods ; Humans ; Hypoglycemic Agents ; metabolism ; Protein Binding ; Serum Albumin ; metabolism ; Sulfonylurea Compounds ; metabolism

OBJECTIVETo observe the effect of Xiaoyao Nose Drops (XYND) in stopping episode of migraine.

METHODSAdopted was the randomized double-blinded placebo-controled method, with the 126 patients equally assigned to two groups, the treated group treated with XYND and the control group with placebo, and the therapeutic course was 30 days for all. The clinical total effective rate and effect in alleviating headache were observed. And hemorrheological parameters as well as the blood flow of the anterior cerebral artery (ACA), median cerebral artery (MCA) and posterior cerebral artery (PCA) were measured before and after treatment.

RESULTSThe clinical total effective rate and headache alleviating rate in the treated group were 93.33% and 96.67% respectively, while those in the control group were 18.33% and 20.00% respectively, and comparison between the two groups showed significant difference (all P < 0.01). The blood viscosity (high, middle and low shear), plasma viscosity and fibrinogen got lowered significantly in the treated group after treatment, showing significant difference in comparison either with those before treatment or with those in the control group after treatment (P < 0.05 or P < 0.01). The velocity of blood flow in all cerebral arteries lowered significantly, also showing significant difference in comparison either with those before treatment or with those in the control group after treatment (P < 0.05 or P < 0.01).

CONCLUSIONXYND is effective in stopping the headache of patients with migraine, and worth applying in clinical practice.

Administration, Intranasal ; Adolescent ; Adult ; Aged ; Blood Flow Velocity ; drug effects ; Blood Viscosity ; drug effects ; Cerebrovascular Circulation ; drug effects ; Double-Blind Method ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Female ; Fibrinogen ; analysis ; Humans ; Male ; Middle Aged ; Migraine Disorders ; drug therapy

OBJECTIVESTo investigate the expression of Survivin in patients with extrahepatic cholangiocarcinoma (EHCC) and its relationship with clinicopathological features of EHCC, and the correlation between the expression of Survivin and lymph node micrometastasis, tumor markers, and the prognosis of EHCC.

METHODSThe expression of Survivin protein in paraffin-embedded specimens of 59 patients with EHCC and their 20 para-carcinoma tissues were evaluated by S-P method of immunohistochemical staining. The correlation between the expression of Survivin and the lymph node micrometastasis, clinicopathological features of EHCC and the prognosis of EHCC were analyzed.

RESULTSThe positive expression rate of Survivin protein was 67.8% (40/59) in paraffin-embedded specimens of 59 patients with EHCC and was 20.0% (4/20) in para-carcinoma tissues, and difference between carcinoma tissues and para-carcinoma tissues was significant (P<0.01). Histological differentiation in EHCC had a negative correlation with the expression of Survivin protein, while the expression of Survivin protein in EHCC had a positive correlation with TNM of EHCC, lymphatic vessel infiltration, lymph node metastasis and perineural invasion (P<0.05). The serum CA19-9 levels in the positive group with expression of Survivin protein was (290,300+/-55 500) U/L and was obviously higher than that in the negative group [(113,300+/-31,400) U/L, P<0.05]. The mean survival time of the patients with negative expression of Survivin protein was higher than that of the patients with positive expression (43.5 vs. 21.1 months, P<0.01). Screened to significance univariate, the multivariate analysis through Cox proportional hazard model analysis showed that lymph node metastasis, residual tumor margins, and expression of Survivin protein were independent prognosis factors of the patients with EHCC (P<0.05, P<0.01, P<0.01).

CONCLUSIONSThe expression of Survivin protein in EHCC has a negative correlation with histological differentiation, while has a positive correlation with lymphatic vessel infiltration and serum CA19-9 concentrations. The expression of Survivin protein maybe an independent prognosis factor of the patients with EHCC.

Adult ; Aged ; Aged, 80 and over ; Bile Duct Neoplasms ; metabolism ; pathology ; Bile Ducts, Extrahepatic ; Cholangiocarcinoma ; metabolism ; pathology ; Female ; Follow-Up Studies ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Lymphatic Metastasis ; Male ; Middle Aged ; Prognosis

BACKGROUNDGlaucoma can cause progressive damage to retinal ganglion cells. These cells can be classified as cells projecting to the superior colliculus and melanopsin-containing retinal ganglion cells, which project to the suprachiasmatic nucleus. This study was to investigate the effects of chronic intraocular pressure elevation on melanopsin-containing retinal ganglion cells in rats.

METHODSChronic intraocular pressure elevation was induced in one eye of adult Wistar rats by cauterization of three episcleral veins. Intraocular pressure was measured at different intervals with a rebound tonometer. Superior collicular retinal ganglion cells were retrogradely labeled from the superior colliculus with Fluorogold. Melanopsin-containing retinal ganglion cells were visualized by free-floating immunohistochemistry on whole-mount retinas. The number of labeled superior collicular and melanopsin-containing retinal ganglion cells were counted in the sample areas on flat-mounted retinas.

RESULTSCompared with contralateral control eyes, the numbers of both superior collicular and melanopsin-containing retinal ganglion cells were significantly reduced after 12 weeks of experimental intraocular pressure elevation ((2317.41 +/- 29.96)/mm(2) vs (1815.82 +/- 24.25)/mm(2); (26.20 +/- 2.10)/mm(2) vs (20.62 +/- 1.52)/mm(2), respectively). The extent of cell loss of the two types of retinal ganglion cells was similar. However, no morphologic changes were found in melanopsin-containing retinal ganglion cells.

CONCLUSIONBoth melanopsin-containing and superior collicular retinal ganglion cells were damaged by chronic ocular hypertension, indicating that glaucomatous neural degeneration involves the non-image-forming visual pathway.

Animals ; Disease Models, Animal ; Glaucoma ; pathology ; Intraocular Pressure ; Male ; Rats ; Rats, Wistar ; Retinal Ganglion Cells ; pathology ; Rod Opsins ; analysis