Objective To investigate the serovar distribution of Chlamydia trachomatis (Ct) isolated from male patients with urethritis in sexually transmitted disease (STD) clinic.Methods Urine specimens were collected from male patients with urethritis in STD clinic at Hospital of Dermatology,Chinese Academy of Medical Sciences between January 2013 and December 2013.Fluorescence-based quantitative PCR was performed to detect Ct DNA in these specimens.DNA was extracted from Ct-positive urine specimens,and nested PCR was conducted to amplify the VS1-VS2 regions of the outer membrane protein A (ompA) gene,followed by gene sequencing.The resulting sequences were aligned to reference sequences by the DNAStar5.0 software to determine Ct serovars.Results A total of 432 urine specimens were collected,and 33.1％ (143/432) of them were positive for Ct.The VS1-VS2 regions of the ompA gene were amplified from 127 out of the 143 Ct-positive specimens,but not from the other 16 specimens.Nine serovars were identified by gene sequencing among the 127 specimens,including serovar E (29 strains,22.83％),F (28 strains,22.05％),D (19 strains,14.96％),G (16 strains,12.60％),J (16 strains,12.60％),K (8 strains,6.30％),H (5 strains,3.94％),I (3 strains,2.36％) and B (3 strains,2.36％),and Ct serovars E,F,D,J and G accounted for 85.02％ among all the strains.Synonymous mutations were identified in 14 out of the 127 strains when compared with reference strains.Conclusions E,F,D and G serovars were the main Ct serovars in male patients with urethritis in STD clinic.The proportion of Ct serovar E strain was decreased,but that of serovar J strain was increased compared with 20 years ago.

Objective To investigate the protective effects of the scallop skirt glycosaminoglycan (SS-GAG) in vitro on the endothelial cell damaged by oxygen free radical and the mechanism of its anti-atherosclerotic effects. Methods The endothelial cells of human umbilical vein (HUVEC)were cultured in vitro, and the damage model of endothelial cells was established by oxygen free radicals . After the damage of HUVEC by Fenton reaction, the effects of SS-GAG on the proliferative activity of HUVEC were observed by the methods of MTT chroma-tometry ,the influence of SS-GAG on lactate dehydrogenase (LDH) release was studied by chromatometry and the effects of SS-GAG on secretion of endothelins and angiotensin Ⅱ were observed by the methods of radio-immunity. Results Compared with the control group, proliferation activity of damage group cell lowered obviously ,LDH release in culture media,and the excretion of endothelins and angiotensin Ⅱ in HUVEC rised obviously. In the groups pre-treated with SS-GAG, cell proliferation activity, the LDH release and the excretions of ET and Ang Ⅱ showed obvious differences compared with model group. Conclusion SS-GAG reduce the damage of endothelial cell caused by oxygen free radicals, restrain the secretion ofendothelins and angiotensin Ⅱ . These results indicate that SS-GAG has protective effects on the endothelial cells damaged by oxygen free redicals and the anti-atherosclerotic mechanism of SS-GAG was related to its protective effects on the endothelial cells.

Phosphoethanolamine N-methyltransferase (PEAMT) is a key enzyme that catalyzes the synthesis of phosphocholine, which is an important precursor of phosphatidylcholine and glycine betaine. A 1249bp 5'-flanking region of phosphoethanolamine N-methyltransferase gene was isolated by anchored PCR, based on the cDNA sequence of PEAMT from halophyte Salicornia europaea. The transcription start site was identified as A and localized at 301bp upstream of the ATG according to the results of RLM-RACE. In SePEAMT promoter region, many potential cis-acting elements were predicted by PlantCARE and PLACE programs. Aside from the basal transcriptional elements TATA-box and CAAT-box, some stress-responsive motifs such as ABRE, HSE and LTR were found. In addition, some pollen-specific activation-related elements were also present in this region. Binary expression vector was constructed by fusing SePEAMT promoter with GUS gene and designated as pPro. The pPro was transferred into tobacco by Agrobacterium-medicated transformation and transient GUS expression analysis indicated that SePEAMT promoter could drive strong GUS expression.

Objective To establish two nested PCR assays for detection of Trichomonas vaginalis in urine samples from male patients with urethritis,and to evaluate their diagnostic value.Methods One thousand and eighty-eight male patients with urethritis were enrolled from sexually transmitted disease (STD) clinic in the Hospital of Dermatology,Chinese Academy of Medical Sciences and Peking Union Medical College between April 2011 and December 2013.Urethral swabs were collected followed by smear testing,wet mount microscopic examination of Trichomonas vaginalis,and cultivation of Neisseria gonorrhoeae.Urine specimens were also obtained from these patients followed by DNA extraction.Two nested PCR assays were developed and performed to amplify the repeat genomic sequence and β-tubulin gene of Trichomonas vaginalis.Results Trichomonas vaginalis was detected in none of these swab specimens by wet mount microscopy,but in 29 (2.67％) of the urine specimens by either of the two nested PCR assays.Moreover,the positive specimens detected by the two nested PCR assays were completely consistent.Conclusion Compared with wet mount microscopy,nested PCR has higher sensitivity and specificity in detection of Trichomonas vaginalis in urine samples from male patients.

OBJECTIVETo study the role of procalcitonin (PCT) in the diagnosis of acute pyelonephritis (APN) in children.

METHODSRetrospective analysis was performed on the clinical records of children aged under 3 years who were diagnosed with primary urinary tract infection (UTI) from September 2011 to February 2012. These children were divided into those with upper UTI (UUTI) (APN) and those with lower UTI (LUTI) (non-APN) based on 99mTc-dimercaptosuccinic acid (DMSA) renal scan results as a gold standard. The UUTI and LUTI groups were compared in terms of serum levels of PCT and C-reactive protein (CRP). Receiver operating characteristic (ROC) curves were drawn to evaluate the diagnostic values of serum PCT and CRP.

RESULTSSixty-five children with UTI, including 39 cases of APN and 26 cases of LUTI, were included in this study. The APN cases had significantly higher serum levels of PCT (3.08 ng/mL vs 0.37 ng/Ml; P<0.01) and CRP (6.25 mg/L vs 3.01 mg/L; P<0.01) than the LUTI cases. The sensitivity and specificity of serum PCT level for APN were 84.6% and 88.5%, respectively, with an area under the ROC curve (AUC) of 0.873 (95%CI=0.781-0.965) and an optimal threshold point of 1.03 ng/mL. The sensitivity and specificity of serum CRP level for APN were 71.8% and 69.2%, respectively, with an AUC of 0.735 (95%CI=0.612-0.858) and an optimal threshold point of 3.91 mg/L.

CONCLUSIONSAs a result of its high sensitivity and specificity for the disease, serum PCT can be used as a marker in the early diagnosis of APN in children.

C-Reactive Protein ; analysis ; Calcitonin ; blood ; Calcitonin Gene-Related Peptide ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Protein Precursors ; blood ; Pyelonephritis ; blood ; diagnosis ; ROC Curve ; Sensitivity and Specificity

Objective To investigate the effect of nicotine on proliferation and chemosensitivity of A549 cells in vitro. Methods A549 cells was assessed by MTT assay to measure cell proliferation and was assessed by RT-PCR tomeasure chemosensitivity. Results 0.01~100μmol/L nicotine could promote the proliferation of A549 cells, the most marked proliferation at 1μmol/L, compared with the control group, the activity of A549 cells was increased by 1.85 times (P<0.01). When the concentration of nicotine above 1μmol/L, the proliferation of A549 cells had an decreasing tendency. When the concentration above 1000μmol/L, the proliferation of A549 cells can be inhibited. Nicotine can also reduce chemosensitivity of A549 cells to 5-FU, with the addition of nicotine, A549 cells survival rate increased significantly, the most marked at 1μmol/L, compared with the control group, the inhibitory rate of A549 cells was 9 % (P< 0.01). Nicotine significantly increased the expression level of α7 nAChR in A549 cells and decreased the expression of PTEN , in a concentration dependent manner. Compared with the control group, 1μmol/L of nicotine could increase the expression levels of α7 nAChR by 3.4 fold, and decrease the expression levels of PTEN by 60.36 % (P< 0.01). Conclusion Nicotinecan promote the growth of A549 cells and reduce chemosensitivity of A549 cells to 5-FU.

OBJECTIVE: To investigate the expression pattern of adapter protein with a Src-homology 2 domain (SH2B1), the suppressor of cytokine signaling-3 (SOCS3), protein-tyrosine phosphatase 1B (PTP1B) and neturopetide Y (NPY) in obese and normal mice hypothalamus and its relation with serum leptin and insulin levels. METHODS: The obesity animal model was prepared with healthy C57/bl6 mice. Lee's index and Homeostasis model assessment-insulin resistance (HOMA-IR) were calculated. The mRNA levels of SH2B1, SOCS3, PTP1B and NPY were measured by fluorescent quantitation RT-PCR. The SH2B1 and NPY protein expressions were detected by Western blot. RESULTS: Compared with the normal mice of the same age, SH2B1 mRNA expression in the obese mice hypothalamus decreased. SOCS3 and PTP1B mRNA expression increased. Western blot showed that SH2B1 protein expression decreased, while NPY protein expression increased in the obese mice. Linear correlation analysis showed that the serum leptin and fasting insulin levels were negatively correlated with SH2B1mRNA expression and positively correlated with SOCS3 and PTP1B mRNA expression. CONCLUSION SH2B1, SOCS3, PTP1B and NPY are key factors for obesity development.
Adaptor Proteins, Signal Transducing ; metabolism ; Animals ; Hypothalamus ; metabolism ; Insulin ; blood ; Insulin Resistance ; Leptin ; blood ; Mice ; Mice, Inbred C57BL ; Neuropeptide Y ; metabolism ; Obesity ; metabolism ; Protein Tyrosine Phosphatase, Non-Receptor Type 1 ; metabolism ; RNA, Messenger ; Suppressor of Cytokine Signaling 3 Protein ; Suppressor of Cytokine Signaling Proteins ; metabolism

PURPOSE: To evaluate the characteristic sequential MRI findings of cortical laminar necrosis. MATERIALS AND METHODS: We retrospectively reviewed the MRI findings of 11 patients with clinical signs of hypoxic brain damage who showed findings of cortical laminar necrosis with definite time of onset. Three were men and eight were women; they were aged between 27 and 74 (mean 59.3)years. All patients underwent imaging with a 1.0-T MagnetomImpact(Siemens) ; follow-up MR examinations were performed in five. RESULTS: The watershed zones in the parietooccipital, frontoparietal and temporoparietal cortex were involved in eight cases, whereas the other areas involved were the frontal lobe in two cases and the temporal lobe in one. In one case, MRI obtained two days latershowed brain swelling demonstrating obliteration of cortical sulci and high signal intensity of subcortical whitematter on T2WI. In five cases, MRI obtained between two and three weeks later showed gyriform high signalintensity along the cortex on T1WI, cortical high and subcortical low signal intensities on T2WI in four cases,and gyriform enhancement on gadolinium-enhanced scans in three cases. MRI obtained between three and four weeks later in three cases showed subcortical high signal intensity on T2WI in two cases and gyriform cortical highsignal intensities on T1WI and gyral enhancement in all cases. MRI obtained after 50 days in four cases includingtwo of follow-up MR, showed cortical gyriform high signal intensity on T1WI in all cases and subcortical high signal intensity on T2WI and mild gyriform enhancement on gadolinium-enhanced scans in three cases. In twofollow-up studies, the lesions had become more discrete and larger. CONCLUSION: Cortical laminar necrosis due tohypoxic brain damage shows relatively characteristic MR findings according to the stage. Therefore, MR imaging seems to be useful diagnostic tool for the evaluation of cortical laminar necrosis due to hypoxic brain damage.
Brain ; Brain Edema ; Female ; Follow-Up Studies ; Frontal Lobe ; Humans ; Hypoxia, Brain ; Magnetic Resonance Imaging* ; Male ; Necrosis* ; Retrospective Studies ; Temporal Lobe

OBJECTIVETo investigate the efficiency of multi-round fluorescence in situ hybridization (FISH) and its influencing factors in preimplantation genetic diagnosis (PGD).

METHODSA total of 48 couples accepted PGD because of various reasons: 24 with Robertsonian translocations, 16 with reciprocal translocations, 2 with pericentric inversions, one with advanced maternal age who had a previous liveborn of Down syndrome, 3 suffered from sex chromosome abnormalities and 2 repeated spontaneous miscarriages. After 72 retrieval cycles, 432 cleavage stage embryos with more than six cells were biopsied on day three. Only intact nuclei (396) were hybridized in order to verify the chromosomal status of the individual embryos. If previous FISH has failed to give conclusive results while the nuclei remained undamaged, the nuclei were hybridized once again. A total of 870 times of hybridization were conducted to 396 nuclei. Signal identification rates of each round as well as the influence of different probes to the hybridization efficiency were compared. Factors leading to inconclusive FISH results were analyzed as well.

RESULTSFive hundred and thirty five out of 870 hybridizations gave identifiable signals (61.5%). The second and third round FISH showed the best signals with an identification rate of 71.8% and 77.4%, respectively, which were significantly higher than those of the first round (52.8%, P < 0.01), the fourth round (55.8%, P < 0.05, P < 0.01), the fifth round (54.5%, P < 0.05) and the sixth round (27.3%, P < 0.01). The identification rate of centromere specific probe signals (CEP group) was 80.3% and the former three rounds in this group got the best quality of signals with an identification rate of 85.7%, 85.1% and 88.0%, respectively, which was significantly higher than that of the latter three rounds. The identification rate of other probe was much lower than with the CEP probe (55.2% vs. 80.3%, P < 0.01) and the best quality of signal in this group was achieved in the fifth round (72.7%), followed by the second round (66.1%) and the third round (63.8%). The identification rate of the first round (50.3%) and the sixth round (22.2%) were significantly lower compared with the second round (P < 0.01). During the 6 rounds of FISH, 335 hybridizations did not give conclusion results (38.5%, 335/870). The main cause of unidentification was weak signals (20.9%, 182/870). Other common factors included background interference (7.6%, 66/870) and failed hybridization (6.1%, 53/870). Rare causes included nucleus damage (1.8%, 16/870), nucleus loss (1.1%, 10/870) and signal split/overlap (0.9%, 8/870).

CONCLUSIONMulti-round FISH can improve the utility of single nucleus in PGD and the former three rounds have the highest efficiency. The hybridization effect of CEP is better than other probe. Poor signal quality is the common cause of unidentification results.

Female ; Genetic Testing ; methods ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Male ; Pregnancy ; Preimplantation Diagnosis ; methods ; Prenatal Diagnosis ; Translocation, Genetic

Monitoring the long-term radiotherapy-associated molecular changes in low-grade gliomas (LGGs) facilitates the understanding of LGG response to radiotherapy. In this study, we used immunohistochemistry to analyze the expression of Ki-67, tumor protein P53 (TP53), P21, and P27 in 8 paired WHO grade II astrocytoma samples. The interval between radiotherapy (RT) and the second surgery was more than 3 months in all cases. The average Ki-67 labeling index (LI) was 5.3% in pre-RT samples and 11.54% in post-RT samples. Ki-67 LI was higher in the primary tumors that underwent malignant transformation observed at the second surgery after radiation. Post-RT Ki-67 LI decreased in 2 cases with an interval of less than 12 months between RT and the second surgery. TP53 expression was found in 3 out of 4 pre-RT samples with malignant transformation and in 1 out of 4 pre-RT samples without malignant transformation. Post-RT TP53 increased in 2 cases in which increased expression of P21 or P27 was also observed. Our study suggests that radiotherapy can inhibit WHO grade II astrocytoma proliferation as reflected by Ki-67 LI, but the effect attenuates with time. In addition, there is a tendency of malignant transformation for WHO grade II astrocytomas with a high Ki-67 level or TP53 expression in initial samples.
Adult ; Astrocytoma ; metabolism ; pathology ; radiotherapy ; surgery ; Brain Neoplasms ; metabolism ; pathology ; radiotherapy ; surgery ; Cell Proliferation ; radiation effects ; Cell Transformation, Neoplastic ; radiation effects ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Female ; Humans ; Immunohistochemistry ; Ki-67 Antigen ; metabolism ; Male ; Middle Aged ; Neoplasm Grading ; Tumor Suppressor Protein p53 ; metabolism