Objective To evaluate the performance of a DNA microarray method for detecting HBV antiviral drug-resistant mutations. Methods Two hundred and twenty four serum samples from patients with CHB were tested in parallel by DNA microarray and direct sequencing for the mutations within the HBV reverse transcriptase (rt) region, which included rtL180, rtA181, rtM204 and rtN236. Samples with discrepant results were retested by clonal sequencing. Results Complete concordance between DNA microarray and direct sequencing results was observed in 214 out of 224 samples (95. 5% ). The presence of mixed viral populations in the other 10 samples detected by DNA microarray but not by direct sequencing was confirmed by clonal analysis. The DNA microarray could detect minor viral populations which constituted 5.0%-15. 0% of the total viral load. Conclusion DNA microarray is highly consistent with direct sequencing in detecting HBV mutations conferring drug resistance and more sensitive in detecting mixed mutant and wild-type sequences than direct sequencing, which makes it a useful tool for early detection of drug resistance early.

Background The ocular nerve of trigeminus is the sensation and nutrition nerve of cornea.Whether trigeminal neuralgia affect the function of ocular surface and the morphology of corneal nerve plexus or not is below understanding.Confocal microscope is a non-invasive method for in vivo corneal examination.Objective This study was to analyze the ocular surface findings and observe the morphology and density of corneal nerve under the confocal microscopy in patients with trigeminal neuralgia.Methods Thirty-three eyes of 33 patients with trigeminal neuralgia were collected from the Department of Pain Management in Henan Provincial People's Hospital.The corneal perceptual sensitivity was examined using corneal aesthesiometer,and the function of lacrimal secretion (Schiemer Ⅰtest),tear break-up time (BUT) were performed to evaluate the influence of trigeminal neuralgia on ocular surface.The change of corneal nerve was observed under the confocal microscopy.The fellow eyes served as controls.The informed consent was obtained from the subjects before any examination.Results The fiber length of corneal perceptual sensitivity was (54.348±6.793)mm and (55.217±6.480)mm in trigeminal neuralgia group and control group without a significant difference between them (t=0.641,P=0.528).No significant differences were found in the mean value of Schiemer Ⅰtest (9.390±6.583mm vs 9.300±5.295mm) and BUT result (6.09±4.177s vs 6.13±4.799s) between trigeminal neuralgia and control group(t=0.070,P=0.945;t=-0.085,P=0.933).The densities value of corneal subepithelial nerve plexus at the nasal,temporal,superior,inferior and central area was insignificantly changed between trigeminal neuralgia group and control group(P=0.840,0.459,0.268,0.120,0.607).Tenuous,bending and circling nerve fibers were seen in corneal stroma under the confocal microscope,while the nerve fibers were strict in controls.Conclusion Trigeminal neuralgia does not dramatically affect eye surface function and corneal subbasal nerve plexus density,but corneal nerve fibers with trigeminal neuralgia are more bending than normal people.

Objective To retrospectively analyse the anterior and posterior surgical approaches in treatment of unstable burst thoracolumbar fractures and compare radiographic measurement parameters of beth surgical techniques so as to provide references for surgical treatment of such kind of fracture. Methods The study selected 41 patients with unstable thoracolumbar fracture treated with either anterior neurodecomprossion and fixation (n=19) or posterior reposition and internal fixation by pedicle screw (n=22) from January 2003 to December 2005. All patients were followed up for 24-48 months ( mean 38 months) and divided into anterior approach group and posterior approach group. Sagittal alignment was assessed by the Cobb angle depending on lateral radiographs. Results The Cobb angle of the anterior approach group was average 27.3°on admission but 3.1°postoperatively and 4.6° at follow-up; while the Cobb angle of posterior approach was average 26.1° on admission, 3.0°postoperatively and 12.5°at follow-up. There was no statistical difference between Cobb angle on admission and postoperative one (P>0.05) but showed significant differences between them at follow-up ( P<0.01). Conclusion The anterior surgical approach can consistently yield better maintenance of kyphotic correction compared with the posterior surgical approach.

Objective To explore the clinical application value of left atrial function with feature tracking cardiac magnetic resonance imaging (FT-CMR) by evaluating preliminarily left atrial strain and strain rate in patients with atrial fibrillation. Methods Thirty patients with paroxysmal atrial fibrillation, thirty patients with persistent atrial fibrillation and twenty-two healthy subjects were enrolled. All the subjects underwent cardiac magnetic resonance imaging with the real steady-state free precession(SSFP) sequence. FT-CMR parameters included left atrial strain and strain rate parameters, left atrial volume and function parameters were detected by using offline cardiovascular analysis software, respectively. Left atrial strain and strain rate parameters included left atrial total strain(Εs), passive strain(Εe), active strain(Εa), peak positive strain rate(SRs), peak early negative strain rate(SRe)and peak late negative strain rate(SRa). Volume and function parameters included maximum of left atrial volume(LAVmax), minimum of left atrial volume(LAVmin), total left atrial emptying fraction(LATEF), passive left atrial emptying fraction(LAPEF)and active left atrial emptying fraction(LAAEF). The differences in the general data among the paroxysmal atrial fibrillation group, the persistent atrial fibrillation group and the control group were compared by usingχ2 test or ANOVA analysis. The differences in all parameters between the atrial fibrillation group and the control group, the paroxysmal atrial fibrillation group and the persistent atrial fibrillation group were compared by using independent t test. Left atrial strain and strain rate parameters on an intra-observer and inter-observer were determined by intraclass correlation coefficient(ICC)analyses. Results Compared to control group, LAVmax and LAVmin in atrial fibrillation group were significantly increased(t=9.737,7.889,P<0.001);The LATEF and LAPEF had no significant difference, the LAAEF in two groups had statistically significant difference(t=-4.762,P<0.001).The absolute value of Es, Ee, Ea, SRs, SRe, SRa in atrial fibrillation group were significantly reduced than in control group(t=-7.732,-6.610,-6.493,-7.546, 6.864, 5.917,P<0.001). Compared with paroxysmal atrial fibrillation group, LAVmax and LAVmin in persistent atrial fibrillation group were increased obviously, LATEF and LAPEF were significantly decreased, and the differences were statistically significant(t=-4.575,-5.524, 4.002, 4.028,P<0.001).The LAAEF in two groups had no statistically significant difference. Compared with strain and strain rate in two groups, absolute value of Es, Ee, Ea, SRs, SRe, SRa in persistent atrial fibrillation group significantly decreased than in paroxysmal atrial fibrillation(t=4.310, 3.128, 4.465, 5.496,-3.290,-3.863,P<0.001). The intra-group and inter-group had well correlation coefficients between the observers in the left atrial strain and strain rate parameters of the subjects(ICC=0.85—0.94,0.81—0.90). Conclusions FT-CMR technique can be used to assess the left atrial strain and strain rate in patients with atrial fibrillation;Left atrial reservoir, conduit and booster-pump functions in patients with atrial fibrillation were impaired. Patients with persistent atrial fibrillation had worse left atrial function throughout the entire cardiac cycle compared with those with paroxysmal atrial fibrillation.

OBJECTIVETo study the effect of Guishen Pill (GSP) on expression levels of Oct-4, MVH, and Egr-1 in mice with diminished ovarian reserve (DOR).

METHODSTotally 40 female C57BL/6J mice were randomly divided into 4 groups, the normal control group, the model group, the GSP group, and the dehydroepiandrosterone (DHEA) group, 10 in each group. Pregnant mare serum gonadotropin (PMSG), human chorionic gonadotropin (HCG), and prostaglandin F2α (PGF2α) were sequentially administrated to produce superovulation. The DOR model was established by exposing to ozone inhalation. Mice in the GSP group were intragastrically administered with GSP at 0.3 mL. Those in the DHEA group were intragastrically administered with DHEA at 0.3 mL. Equal volume of normal saline was intragastrically administered to mice in the normal control group and the model group. All mice wer treated for 21 days. Serum levels of estrogen (E2), progestogen (P), and anti-Müllerian hormone (AMH) were measured by ELISA. Changes of Oct-4, anti-AMH, and early growth response gene-1 (Egr-1) mRNA in ovaries were dtected by Real-time PCR.

RESULTSCompared with the model group, serum levels of E2, P, and AMH, as well as contents of estrogen receptor (ER), progestogen receptor (PR), MVH, and Oct-4 mRNA significantly increased in the GSP group and the DHEA group (P < 0.05).

CONCLUSIONGSP could improve expression levels of Oct-4, MVH, and Egr-1 mRNA in DOR mice and their ovarian function.

Animals ; Anti-Mullerian Hormone ; metabolism ; Dehydroepiandrosterone ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Early Growth Response Protein 1 ; metabolism ; Estrogens ; Female ; Mice ; Mice, Inbred C57BL ; Octamer Transcription Factor-3 ; metabolism ; Ovarian Reserve ; Ovary ; Pregnancy ; Receptors, Estrogen ; metabolism ; Superovulation

AIM:To investigate the effect of Eph receptor A2 (EphA2) on drug resistance of colorectal carci-noma cells and its possible mechanisms .METHODS:Real-time PCR and Western blot were used to detect the expression of EphA2 at mRNA and protein levels in LoVo and LoVo/5-FU cells.EphA2 siRNA was transfected to down-regulate the EphA2 expression in LoVo/5-FU cells, and the drug sensitivity was calculated by CCK-8 assay.Meanwhile, cell migration and invasion were measured by wound healing assay and Transwell assay , and the protein levels of E-cadherin,β-catenin, N-cadherin, vimentin, Notch and Snail were determined by Western blot .RESULTS: The expression of EphA2 at both mRNA and protein levels was significantly up-regulated in LoVo/5-FU cells (P<0.05).Knockdown of EphA2 suppressed the cell viability, and migration and invasion abilities , but promoted drug sensitivity of LoVo/5-FU cells.Up-regulation of E-cadherin and β-catenin, and down-regulation of N-cadherin and vimentin were observed , indicating that the epithelial-mesenchymal transition ( EMT) process was suppressed .Knockdown of EphA2 decreased the expression levels of Notch and Snail.CONCLUSION:Down-regulation of EphA2 partly reverses drug resistance of LoVo/5-FU cells.The mechanism may be related to suppressing cell growth , migration, invasion and EMT process via Notch/Snail signaling pathway .

OBJECTIVETo examine the effect of exposure to extremely low-frequency electromagnetic fields (ELF EMFs) on the liver function of workers.

METHODSThe workers in a factory were selected as subjects, and the recent physical examination data of these workers were collected. The workers aged 20∼40 years and with more than 2 years' working experience were included for analysis; considering the intensity of electromagnetic field, the workers exposed to less electromagnetic radiation were assigned to exposure I group (n = 123), those exposed to more electromagnetic radiation to exposure II group (n = 229), and those not exposed to electromagnetic radiation to control group (n = 212). There were no significant differences in sex, age, height, and body weight between the three groups (P > 0.05). Physical examination, including measurements of direct bilirubin (DBil), alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), γ-glutamyl transpeptidase (GGT), and albumin, was performed in a health examination center. The intensity of electromagnetic field was measured by EFA-300 power frequency electromagnetic field analyzer, and the intensity of noise by AWA5610D integrating sound level meter.

RESULTSThe intensities of electric field and the magnetic field in exposure II group were significantly higher than those in the exposure I group. The levels of ALT, ALP, AST, GGT and albumin in exposure II group were significantly higher than those in exposure I group and control group. However, the level of direct bilirubin in exposure II group was significantly lower than that in exposure I group and control group.

CONCLUSIONOccupational exposure to ELF EMFs may affect human liver function.

Adult ; Alanine Transaminase ; blood ; Aspartate Aminotransferases ; blood ; Bilirubin ; blood ; Electromagnetic Fields ; adverse effects ; Female ; Humans ; Liver ; physiopathology ; Male ; Occupational Exposure ; adverse effects ; Young Adult

Objective To construct recombinant adeno-associated virus and lentivirus carrying siRNA of TIMP-1 and to investigate the efficiency of infection and short-term inhibitory effect of TIMP-1 gene expres-sion on rat hepatic stellate cells. Methods One pair of siRNA which could effectively inhibit expression of the TIMP-1 gene in HSC-T6 was screened and cloned into AAV vector and lentiviral vector to construct the recombinant AAV/siRNA-TIMP-1/GFP and Lenti/siRNA-TIMP-1/GFP. AAV/GFP and Lenti/GFP as neg-ative control were also obtained. Experiments were assigned to five groups: AAV/siRNA-TIMP-1/GFP, AAV/GFP, Lenti/siRNA-TIMP-1/GFP, Lenti/GFP group and mock group. Rat HSC-T6 cells were infected by these recombinant viruses at a concentration of MOI by 10. To monitor the efficiency of infection, fluores-cence microscope and flow cytometer were used. After 7 d post-infection, Western blot was used to detect the TIMP-1 protein expression. Results HSC-T6 had no significant changes after infection. The efficiency of infection in AAV/GFP and Lenti/GFP group were 72.7% and 70.0%, AAV/siRNA-TIMP-1/GFP and Lenti/siRNA-TIMP-1/GFP group were 64.58% and 61.86%. The protein expression levels of TIMP-1 in HSC-T6 cells at 7 d post-infection by the recombinant AAV and Lentivirus were decreased 40.0% compared with those in mock control and normal HSC-T6 (P<0.05). Conclusion Recombinant AAV/siRNA-TIMP-1/GFP and Lenti/siRNA-TIMP-1/GFP could effectively infect HSC-T6 with similar efficiency and suppress the expression of TIMP-1 in rat HSC-T6 remarkably.

OBJECTIVETo explore the effect of seawater immersion on serum osmotic pressure and electrolytes balance following chest trauma in dogs.

METHODSTwenty-five healthy adult dogs were used in the experiment. A canine model of right open pneumothorax was established by chest puncturing on all animals. Animals were divided into three groups: a control group (n=10) with chest trauma without any immersion; a seawater group (n=10) immersed in seawater after chest trauma and a normal saline group (n=5) immersed in normal saline solution following chest trauma. Blood samples were taken at different time intervals to determine plasma osmotic pressure and electrolytes. The hemodynamic changes were also recorded.

RESULTSMortality in the seawater group was much higher than that of the control group and the normal saline group. The mean survival time in the seawater group lasted only 45 minutes, while in the control group and the normal saline group the average survival time was more than 4 hours (P<0.01). One of the most important causes of death was hypernatremia and high osmolality. Severe electrolytes imbalance was observed in seawater group. Hypernatremia and high osmolality were the most significant factors of high mortality in the seawater group.

CONCLUSIONSSeawater immersion after chest trauma appears to be associated with severe electrolyte imbalance as well as high osmotic pressure. These may be the risk factors leading to fatal outcome.

Animals ; Dogs ; Hemodynamics ; Osmotic Pressure ; Risk Factors ; Seawater ; adverse effects ; Thoracic Injuries ; physiopathology ; Water-Electrolyte Balance ; physiology

OBJECTIVETo analyze the degrees on the epidemic foci of Angiostronglus cantonensis and to explore the measurement methods.

METHODSSnails (Pila gigas) were collected from the spots of Wenzhou, Cangnan, Yongjia, Yueqing in Zhejiang province and Minhou, Changle, Ningde in Fujian province. The snails were examined microscopically in order to calculate their infection rates and the average worm number in the positive snails, then taking the product of multiplication of both values as infestation index.

RESULTSThe infection rates of the epidemic foci were 10.59% (9/85), 60.74% (181/298), 34.96% (79/226), 32.90% (76/231), 57.50% (184/320), 40.00% (82/205), 17.65% (12/68) and the rates of infectivity were 6.57, 183.54, 121.73, 93.45, 276.36, 76.08, 12.65, respectively.

CONCLUSIONThe epidemic foci were divided into five ranks (super, high, mid-range, low and non-epidemic foci) according to the value of infestation index which ranked from > 75, 30-75, 5-29, < 5 to 0.

Angiostrongylus ; pathogenicity ; Animals ; China ; Disease Outbreaks ; Snails ; parasitology ; Strongylida Infections ; epidemiology