1.Detection of high-risk human papillomavirus by multiplex real time PCR
Kelin XIAO ; Huilin WANG ; Guangxing MAI ; Jing MA ; Fan JIANG ; Xiufa ZHANG ; Likuan XIONG ; Hua ZHOU
Chinese Journal of Laboratory Medicine 2011;34(6):534-537
Objective To investigate 13 high-risk types of HPV (HR HPV) infection rates in women with different grades of cervical lesions.Methods A total of 350 women, who were hospitalized in the department of gynecology in Bao′an Maternity & Child health hospital, were enrolled for the study.TCT technology was used to evaluate the cervical epithelium.The group were divided according to the cytology results.Multiplex real time PCR (mRT PCR) was used to detect the viral loads.HR HPV infection rate of different groups were analyzed using χ2 test or Fisher exact test.HR HPV viral loads of patients in different grades of cervical lesion groups were compared using Kruskal-Wallis or Wilcoxon test, and the age distribution of HR HPV positive group and negative group was analyzed by using Wilcoxon test.Results The HR HPV infection rates of NILM, ASCUS, LSIL, HSIL were 3.4% (10/295), 20.0% (7/35), 78.6% (11/14) and 100.0% (6/6), respectively.HR HPV positivity in NILM was lower than ASCUS (χ2=14.43,P<0.01) and LSIL (χ2=107.69,P<0.01), HR HPV positivity in ASCUS was lower than LSIL (χ2=14.76,P<0.01). The median of HR HPV viral loads in NILM, ASCUS, LSIL and HSIL were 4.10 (3.38-6.27), 5.33 (3.63-6.66), 5.77 (4.01-7.01) and 5.58 (4.19-5.85) respectively (copies/ml,lg).Combined ASCUS, LSIL and HSIL groups into cervical lesion group, HR HPV viral load of which was higher than that of NILM (U=43.0, P<0.05).The median Ages of HR HPV positive group and negative group were 36 and 33, respectively.No statistical significance was found between them (U=4 544, P>0.05).Conclusions The present study revealed that HR HPV infection was related to cervical lesion, but there was no correlation between viral load and cervical lesion grade. In additional, no difference in age distribution was found between HR HPV positive group and negative group.
2.Clostridium difficile carriage in infants and the characteristics of isolates
Tianxiang ZHOU ; Zhenhua SHU ; Kelin XIAO ; Zhongxing WANG ; Guangxing MAI ; Yong XIA ; Zhen HUANG ; Huisheng FAN ; Conghui GU
International Journal of Laboratory Medicine 2017;38(13):1735-1737
Objective To investigate the carrying status and characteristics of Clostridium difficile isolated from infants.Methods Two hundred and thirty-eight stool specimens were collected from infant younger than 1 year old,that were hospitalized or outpatient from August to November 2015.Immunochromatography targeted GDH and toxin A&B of C.difficile was used for C.difficile screening,and those positive specimens were inoculated in CDIF and anaerobic culture.C.difficile isolates were genotyped by using slpA sequence typing (slpA ST),and tcdA,tcdB,cdtA and cdtB of C.difficile isolates were detected by PCR.Results Fifty C.difficile strains were isolated from 238 stool samples,and the isolated rates of C.difficile from <3 months,3 months to <6 months,and 6 months to 1 years old groups were 9.3%,17.6% and 27.3%(χ2=6.940,P=0.031<0.05),respectively.52.0%(26/50) of the C.difficile isolates were toxigenic,and 69.2% (18/26) toxigenic isolates harbored tcdA+tcdB+cdtA-cdtB-.Fifty C.difficile isolates were genotyped as 11 slpA STs,slpA ST fr-02 and kr-02 were the commonest genotypes in toxigenic C.difficile isolates;however,that was slpA ST xr-03 in non-toxigenic isolates.Conclusion High C.difficile carriage is found in infants younger than 1 year old,and more than half of C.difficile isolates are toxigenic.Most of toxigenic isolates harbored toxin A and B.The genotype of C.difficile isolates is different between toxigenic isolates and non-toxigenic isolates.
3.Expression of Urotensin II and its receptor on right ventricle in rats of pulmonary hypertension.
Hong HUANG ; Yong-Sheng GONG ; Xiao-Fang FAN ; Liang-Gang HU ; Jian-Feng LUO ; Xiao-Mai WU
Chinese Journal of Applied Physiology 2006;22(1):81-84
AIMTo observe the expression of Urotensin II (U II) and its receptor (UT) on right ventricle in rats with chronic pulmonary hypertension induced by hypoxia and hypercapnia.
METHODSTwenty male SD rats were randomly divided into normal control group (NC) and hypoxia-hypercapnia 4-week group(HH). Mean pulmonary arterial pressure(MPAP) and the weight ratio of right ventricle (RV) to left ventricle plus septum (LV+ S) were calculated separately. U II in plasma was measured using radioimmunoassay. The expression of U II was observed in right ventricle myocytes and right ventricle arteries by immunohistochemistry. The expression of U II mRNA and UT mRNA were observed in right ventricle myocytes and right ventricle arteries by in situ hybridization.
RESULTS(1) The MPAP and RV/LV + S of HH group were higher respectively than those of NC group (P < 0.01, respectively). (2) The plasma U II content of HH group did not increased obviously than that of NC group. (3) The expression score of U II, U II mRNA, UT mRNA by right ventricle myocytes in HH group were higher significantly than those of NC group (P < 0.01 respectively). (4) The average value of integral light density (LD) of U II, U II mRNA, UT mRNA by right cardial arteries in HH group were higher significantly than those of NC group (P < 0.01, respectively).
CONCLUSIONThe expression of U II in right ventricle arteries and right ventricle myocytes increase significantly during the formation of pulmonary hypertension and right ventricle hypertrophy in rats chronically exposed to hypoxia-hypercapnia. These changes indicate that U II might be involved in right ventricle remodeling, which promotes proliferation of cardiac muscle cells.
Animals ; Heart Ventricles ; metabolism ; Hypertension, Pulmonary ; metabolism ; Hypoxia ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, G-Protein-Coupled ; metabolism ; Urotensins ; metabolism
4.Research on the correlation between lupus erythematosus cell and diseases activity in patients with systemic lupus erythematosus
Mai-Gui YANG ; Yang YANG ; Wen-Ke CAI ; Zhu-Ying ZHANG ; Shan-Luan ZHENG ; Ai-Lin FAN ; Ling QU ; Xiao-Ming WEI ; Xiao-Ke HAO
Chinese Journal of Rheumatology 2003;0(08):-
Objective To observe the morphologic changes of peripheral blood lupus cell in patients with systemic lupus erythematosus and investigate its relationship with disease activity in systemic lupus ery- thematosus.Methods Modified classical blood clotting method to observe the morphological changes of pe- ripheral blood lupus cells in 80 cases with systemic lupus erythematosus.Fifty cases were in active stage,and 30 cases were in stable stage.Comparison of serum autoantibody,complement and SLEDAI were also carried out.Results There was significant association between lupus cells in special morphous and autoantibody, such as anti-double-stranded DNA antibody,anti-nucleosome antibodies,complement C3、C4 and SLEDAI(r= 0.588,P=0.056:r=0.759,P=0.135;r=-0.648,P=0.058;r=-0.589,P=0.057,r=0.686,P
5.Sexual hormone levels in semen and germ cell apoptosis.
Mai-Gui YANG ; Yang YANG ; Ping HUANG ; Xiao-Ke HAO ; Zhu-Ying ZHANG ; Shan-Luan ZHENG ; Ai-Lin FAN ; Guo-Zhou RAO ; Xiao-Ming WEI
National Journal of Andrology 2006;12(5):432-434
OBJECTIVETo explore the relationship between sexual hormones in semen and germ cell apoptosis in male population.
METHODSSixty-six infertile patients and thirty fertile males were selected randomly. The levels of folicle stimulating hormone ( FSH), prolactin (PRL), luteinizing hormone (LH), and testosterone (T) in semen were measured by ELISA. Terminal deoxynucleotidyl transferase mediated UTP nick end labeling (TUNEL) was used for the detection of germ cell apoptosis.
RESULTSThe levels of FSH, LH, PRL, T in thirty fertile men were (1.63 +/- 0.15) U/L, (2.18 +/- 0.21) U/L, (6.34 +/- 0.30) nmol/L, (1.85 +/- 0.11) nmol/L, respectively, and germ cell apoptosis rate was (4.61 +/- 1.23)%. FSH, LH, PRL, T levels in infertile group were (1.25 +/- 0.18) U/L, (1.76 +/- 0.32) U/L, (5.86 +/- 0.13) nmol/l, (1.45 +/- 0.13) nmol/, respectively, and germ cell apoptosis rate was (18.36 +/- 2.04)%. There were significant differences in all parameters between infertile group and fertile group. The levels of FSH, LH, PRL, T were negatively correlated with germ cell apoptosis rates( r = -0.88, -0.93, -0.90, -0.98). The volume of apoptotic germ cell decreased, and chromatin was compacted to form cell-membrane blebs and apoptotic bodies.
CONCLUSIONLow concentration of sexual hormones may increase the apoptosis of germ cells, which can induce male infertility.
Adult ; Apoptosis ; Case-Control Studies ; Follicle Stimulating Hormone ; metabolism ; Germ Cells ; pathology ; Gonadal Steroid Hormones ; metabolism ; Humans ; Infertility, Male ; metabolism ; pathology ; Luteinizing Hormone ; metabolism ; Male ; Prolactin ; metabolism ; Semen ; metabolism ; Testosterone ; metabolism
6.Sodium nitroprusside facilitates human sperm capacitation and acrosome reaction.
Mai-Gui YANG ; Yang YANG ; Ping HUANG ; Shan-Luan ZHENG ; Ai-Lin FAN ; Xiao-Dong CHENG ; Tie-Cheng ZHOU ; Juan LI ; Zhu-Ying ZHANG ; Xiao-Ke HAO
National Journal of Andrology 2005;11(6):422-425
OBJECTIVETo explore the effect of nitric oxide (NO) on human sperm capacitation and acrosome reaction (AR).
METHODSDifferent concentrations of sodium nitroprusside (SNP) were added to the sperm suspension from 48 healthy fertile men, and the suspension was incubated in 1 x Earle at 37 degrees C for 1 hour. Progesterone was used to induce AR for 15, 30, 45 and 60 min, and then acid phosphatase (ACP) activity in the suspension before and after capacitation and at different time of AR was measured by p-nitrophenyl sodium phosphate assay. In the meantime, sperm motile parameters were assayed by CASA to observe sperm capacitation and AR.
RESULTSACP activity and sperm motile parameters increased in the 50 approximately 100 nmol/L NO concentration group, showed no significant variation in the 150 approximately 200 nmol/L group, and decreased in the 250 approximately 300 nmol/L group.
CONCLUSIONNO can facilitate sperm capacitation, AR and sperm motile parameters in low concentration and suppress them in high concentration. ACP activity assay of sperm is an objective and reliable method to evaluate sperm capacitation and AR in whole sperm population.
Acid Phosphatase ; metabolism ; Acrosome Reaction ; drug effects ; physiology ; Adult ; Dose-Response Relationship, Drug ; Humans ; Male ; Nitric Oxide ; physiology ; Nitric Oxide Donors ; pharmacology ; Nitroprusside ; pharmacology ; Sperm Capacitation ; drug effects ; physiology ; Sperm Motility ; drug effects ; physiology ; Spermatozoa ; enzymology
7.Changes of intermedin/adrenomedullin 2 and its receptors in the right ventricle of rats with chronic hypoxic pulmonary hypertension.
Yong-Sheng GONG ; Xiao-Fang FAN ; Xiao-Mai WU ; Liang-Gang HU ; Chao-Shu TANG ; Yong-Zheng PANG ; Yong-Fen QI
Acta Physiologica Sinica 2007;59(2):210-214
The purpose of the present study was to explore the expression changes of intermedin/adrenomedullin 2 (IMD/ADM2), a novel small molecular bioactive peptide, and its receptors, calcitonin receptor-like receptor (CRLR) and receptor activity modifying proteins (RAMP1, RAMP2, RAMP3) in the right ventricle of rats with chronic hypoxia-induced pulmonary hypertension. Twenty male Sprague-Dawley rats were randomly divided into 4-week hypoxia group and normal control group (each n=10). The rats in hypoxia group were placed in an isobaric hypoxic chamber, in which O(2) content was maintained at 9%-11% by delivering N(2), and CO(2) content was maintained at <3% for 4 weeks (8 h/d, 6 d/week). The rats in the control group were housed in room air. The protein levels of IMD/ADM2 and adrenomedullin (ADM) in blood plasma and right ventricular tissue were measured by radioimmunoassay. The mRNA expressions of IMD/ADM2, ADM and their receptors CRLR, RAMP1, RAMP2, RAMP3 in right ventricular tissue were determined by reverse transcription-polymerase chain reaction (RT-PCR). The results showed that the ratio of right ventricle weight to left ventricle plus septum weight [RV/(LV+S)] and mean pulmonary arterial pressure (mPAP) were higher in hypoxia group than those in the control group (all P<0.01), suggesting that the rat model of pulmonary hypertension was successfully established. However, the mean carotid arterial pressure (mCAP) between the two groups had no significant difference. Compared with that in the control group, ADM contents in plasma and right ventricular tissue in hypoxia group increased by 1.26 and 1.68 folds (all P<0.01), respectively. Likewise, IMD/ADM2 contents in blood plasma and right ventricular tissue in hypoxia group increased by 0.90 and 1.19 folds (P<0.01), respectively, compared with that in the control group. The data of RT-PCR showed that mRNA levels of ADM, IMD/ADM2 and RAMP2 in hypoxia group increased by 155.1% (P<0.01), 80.9% (P<0.01) and 52.9% (P<0.05), respectively, compared with those in the control group. There were no significant differences in mRNA expressions of CRLR, RAMP1 and RAMP3 between the two groups (all P>0.05). Taken together, the results show that the level of IMD/ADM2 increases in the rats with chronic hypoxia-induced pulmonary hypertension.
Adrenomedullin
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metabolism
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Animals
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Calcitonin Receptor-Like Protein
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metabolism
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Heart Ventricles
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metabolism
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Hypertension, Pulmonary
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etiology
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metabolism
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physiopathology
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Hypoxia
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complications
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Male
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Neuropeptides
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metabolism
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Rats
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Rats, Sprague-Dawley
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Receptor Activity-Modifying Proteins
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metabolism
8.Effects and relationship between NO and HIF-1alpha in rats with pulmonary hypertension induced by hypoxia.
Jian-Feng LUO ; Xiao-Mai WU ; Xiao-Fang FAN ; Liang-Gang HU ; Hong HUANG ; Shan-Shan JIA ; Sun Zhong MAO ; Yong-Sheng GONG
Chinese Journal of Applied Physiology 2006;22(4):488-491
AIMTo investigate the expression of hypoxia-inducible factor-1 alpha (HIF-1alpha) in rats with chronic pulmonary hypertension induced by hypoxia and hypercapnia and its relationship with nitric oxide(NO).
METHODSFourty male Sprague-Dawley rats were randomly divided into four groups, normal control group (NC), hypoxia-hypercapnia group (HH), hypoxia - hypercapnia + L-arginine liposome group(HP) and hypoxia-hypercapnia+ N-nitro-L-arginine methylester group (HM). Colorimetric analysis, immunohistochemistry and in situ hybridization were used for detection of NO, HIF-1alpha and constitutive nitric oxide synthase (ecNOS).
RESULTS(1) The mean pulmonary arterial pressure (mPAP) and the weight ratio of right ventricular to left ventricle plus septum (RV/(LV + S)) of HH group were higher than those of NC group (P < 0.05), HP group much lower than HH group (P < 0.01), mPAP of HM higher than HH group ( P < 0.05). 2)0 Contents of NO in plasma and pulmonary tissue homogenates of HH group were much lower than those of NC group (P < 0.01), HP group higher than HH group (P < 0.01). There were no difference between HM group and HH group. (Expression of HIF-1alpha and HIF-1alpha mRNA in pulmonary arterioles of HH group were significantly higher than those of NC group( P < 0.01), HP group lower than HH group (P < 0.01) ,HM group higher than HH group (P < 0.01); Whereas expression of ecNOS and ecNOS mRNA in pulmonary arterioles of HH were lower than those of NC group( P < 0.05, IP group higher than HH group (P < 0.01), HM group lower than HH group (P < 0.05).
CONCLUSIONHIF-1alpha is involved in the pathogenesis of chronic pulmonary hypertension induced by hypoxia and hypercapnia. The protective function of NO in the pathogenesis might be partly depended on its effects on the expression/activity of HIF-1alpha in lung.
Animals ; Hypertension, Pulmonary ; etiology ; metabolism ; Hypoxia ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Male ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley
9.Changes of adrenomedullin 2/intermedin in the lung of rats with chronic hypoxic pulmonary hypertension.
Xiao-fang FAN ; Ping HUANG ; Yong-sheng GONG ; Xiao-mai WU ; Liang-gang HU ; Li-xian TIAN ; Chao-shu TANG ; Yong-zheng PANG
Chinese Journal of Applied Physiology 2007;23(4):467-471
AIMTo investigate the changes and probable roles of adrenomedullin2/intermedin (AIDM2/IMD), a novel micromolecular bioactive peptide, in the lungs of rats with chronic hypoxic pulmonary hypertension.
METHODSTwenty male SD rats were randomly divided into normal control group (NC) and normobaric hypoxia group (4H). The protein levels of ADM and ADM2/IMD) in the plasma and lung were measured by radioimmunoassay and immunohistochemistry. The mRNA expressions of ADM, ADM2/IMD and their receptors C (RLR, RAMP1, RAMP2 and RAMP3 in the lung tissue were determined by reverse transcription-polymerase chain reaction (RT-PCR).
RESULTS(1) The rat model of chronic pulmonary hypertension was confirmed by the increased mean pulmonary arterial pressure (mPAP) and weight ratio of right ventricle to left ventricle plus septum [RV/(LV + S)] in 4H group compared to NC group. (2) The concentrations of ADM in the plasma and lung homogenate of 4H group were 2.3 and 3.2 folds of NC group, respectively (all P < 0.01). The levels of ADM2/IMD were higher 89.6% and 45.0% in the plasma and lung homogenate of 4H group than those of NC group (respectively, P < 0.01, P < 0.05). (3) The mRNA expressions of ADM2/IMD and ADM in the lung of 4H group were up-regulated (respectively, P < 0.01, P < 0.05 vs. NC group). The expressions of CRLR and RAMP1 mRNAs were down-regulated (all P < 0.01 vs. NC group), while the levels of RAMP2 and RAMP3 mRNAs were no significant difference between the two groups. (4) The strong ADM2/IMD immunostaining was detected in the endothelial and adventitial cells of the rat pulmonary arteriole.
CONCLUSIONADM2/IMD, like its paralog ADM, might be closely related to the chronic hypoxic pulmonary hypertension in rats. The disorders of the gene expression and/or the synthesis and metabolism of ADM2/IMD and its receptor CRLR/RAMP1 possibly take part in the pathogenesis of chronic hypoxic pulmonary hypertension in rats.
Adrenomedullin ; metabolism ; Animals ; Hypertension, Pulmonary ; etiology ; metabolism ; Hypoxia ; complications ; metabolism ; Lung ; metabolism ; Male ; Neuropeptides ; metabolism ; Rats ; Rats, Sprague-Dawley
10.Dynamic changes of urotensin II receptor in pulmonary artery and arterioles of rats chronically exposed to hypoxia-hypercapnia.
Yong-Sheng GONG ; Xiao-Fang FAN ; Xiao-Mai WU ; Yu-Qi GAO ; Liang-Gang HU ; Hong HUANG ; Shan-Shan JIA
Chinese Journal of Applied Physiology 2005;21(4):377-381
AIMTo investigate the dynamic changes and functions of urotensin II (U lI) receptor (UT) in pulmonary arteries of rats chronically exposed to hypoxia-hypercapnia.
METHODSIn rats with hypoxia-hypercapnia at 1, 2 and 4 weeks U II receptor binding of pulmonary arteries sarcolemma was determined by radioligand assay. U II mRNA and UTmRNA in various grades of pulmonary arterioles were measured by in situ hybridization.
RESULTS(1) Mean pulmonary pressure (mPAP) and weight ratio of right ventricle to left ventricle and septum (RV/LV + S) of 1-week group were higher than those of normal control (NC) group by 26.2% and 21.6% (P < 0.01), respectively, and 2-week group higher than 1-week group by 22.5% and 14.1% (respectively, P < 0.01). However, no significant changes were found between 4-week and 2-week group. (2) U Il receptor (Bmax) of 1-week group was higher than NC group by 38.8%, 2-week group higher than 1-week group by 23.2%, and 4-week group increased 7.3% compared with 2-week group (respectively, P < 0.01). The UT changes were time-dependent, while the affinity to U II (Kd) was no different among each group. (3) UII mRNA in each grade of pulmonary arterioles of 2-week group and 4-week group were higher than NC group (respectively, P < 0.01), and those of 2-week group were higher than 1-week group by 5.9% (P > 0.05), 16.4% and 9.1% (respectively, P < 0.01), while no differences existed between 2-week group and 4-week group. (4) UT mRNA in each grade of pulmonary arterioles of all hypoxia-hypercapnia groups was higher than NC group (respectively, P < 0.01), and those of two abaxial grade vessels in 1-week group were the highest. No differences existed between 2-week group and 4-week group. (5) The pulmonary vessels remodeling were time-dependently aggravated by hypoxia-hypercapnia.
CONCLUSIONThe dynamic changes of UT in pulmonary arterioles might have important contribution to the development of pulmonary hypertension and pulmonary arteriole remodeling induced by chronic hypoxia-hypercapnia in rats.
Animals ; Arterioles ; metabolism ; physiopathology ; Hypercapnia ; metabolism ; Hypertension, Pulmonary ; metabolism ; physiopathology ; Hypoxia ; metabolism ; Male ; Pulmonary Artery ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Receptors, G-Protein-Coupled ; metabolism