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1.
Chinese Journal of Dermatology ; (12): 176-181, 2019.
Article in Chinese | WPRIM | ID: wpr-745760

ABSTRACT

Objective To compare and analyze the differences in the transcriptomics between mycelium and early yeast phases of Sporothrix schenckii (S.schenckii),and to realize the changes in transcriptome expression profiles during mycelium-to-yeast transformation.Methods A standard strain of S.schenckii (ATCC 10268) was subjected to 96-hour culture with Sabouraud medium at 25 ℃ or 36-hour culture with brain-heart infusion medium at 37 ℃ to obtain the mycelium and yeast form of S.schenckii,and then,their transcriptomes were sequenced.Functional annotation was performed for screened unigenes by comparison using several databases (such as NR,Swiss-Prot,KEGG,COG,KOG,GO and Pfam),coding sequence prediction,and gene expression analysis in each sample.Finally,the differentially expressed genes were subjected to pattern clustering,functional annotation and enrichment analysis.Results A total of 14.76 Gb valid data (clean data) were obtained,and functional annotation results were acquired in 28 094 of 43 863 assembled unigene clusters.Compared with S.schenckii in mycelium phase,there were 10 969 up-regulated genes and 199 down-regulated genes in S.schenckii in yeast phase.These differentially expressed genes were involved in protein phosphorylation,intracellular protein transport,cellular protein modification,small guanosine triphosphate-mediated signal transduction,vesicle-mediated transport,translation,intracellular signal transduction,microtubule formation,adenosine triphosphate synthesis,coupled proton transport and so on.Sixteen genes in the mitogen-activated protein kinase (MAPK) pathway and two-component signaling pathway,which were two important signal transduction pathways involved in fungal morphogenesis,and 16 genes involved in chitin synthesis and metabolism were all confirmed to be up-regulated in S.schenckii in yeast phase.Conclusions Compared with S.schenckii in mycelium phase,great changes in gene expression profiles were observed in S.schenckii in yeast phase.These differentially expressed genes are involved in many functions,suggesting that the dimorphic transition of S.schenckii is regulated by a multi-gene network system.

2.
Article in Chinese | WPRIM | ID: wpr-510615

ABSTRACT

Objective To determine the effects of different doses of cisatracurium on motor e-voked potential of neurosurgery operation.Methods Sixty patients,36 males and 24 females,aged 18 to 65 years,ASA physical status Ⅰ or Ⅱ,scheduled for spinal surgery with motor evoked potential monitoring,were included and randomly assigned to three groups.A single dose of cisatra-curium besilate for injection was given by intravenous injection in 5 s after the induction of general an-esthesia,respectively 0.1 mg/kg (group A),0.1 5 mg/kg (group B)and 0.2 mg/kg (group C).Cas-cade Elite 32 channel monitor was used to monitor MEPs,the electrode was stimulated for once two minutes after given the muscle relaxant,and the leading time of the wave of MEPs was recorded. Cooper’s score was used to evaluate the intubation conditions.Results The appearance time of the wave of motor evoked potentials was significantly longer in group C [(39.60±1.79)min]than that in groups A [(20.10 ± 1.89 )min]and B [(20.50 ± 1.93 )min](P < 0.05 ).The intubation conditions was significantly better in group B (100%)and C (100%)than that in group A (65%)(P<0.05).Conclusion The shortest time to elicit waveform of MEPs using the dose of cisatracurium is 0.1 5 mg/kg at induction of general anesthesia,which is better for tracheal intubation.The dose 0.1 5 mg/kg of cisatracurim is recommended as the initial dose on neurosurgery operation with motor e-voked potential monitoring.

3.
Journal of Clinical Hepatology ; (12): 2016-2020, 2016.
Article in Chinese | WPRIM | ID: wpr-778442

ABSTRACT

Primary biliary cholangitis is an organ-specific chronic cholestatic autoimmune liver disease characterized by intrahepatic cholestasis, presence of anti-mitochondrial antibody in circulating blood, and progressive and non-suppurative damage of small intrahepatic bile ducts, which finally lead to extensive hepatic duct damage, biliary cirrhosis, and even liver failure. This article overviews the advances in the epidemiology, pathogenesis, diagnosis, and treatment of primary biliary cholangitis and points out that the safety and efficacy of the new drugs such as fibrates in the treatment of primary biliary cholangitis need to be confirmed by further studies.

4.
Article in Chinese | WPRIM | ID: wpr-499923

ABSTRACT

Objective To explore the clinical significance of monitoring the femoral artery hemodynamics of acute cerebral hemorrhage patients under general anesthesia by color doppler ultrasound .Methods Totally 30 patients who received emergency craniotomy for intracra-nial hematoma were performed the induction of general anesthesia and endotracheal intubation .The changes of femoral artery peak systolic ve-locity (Vs),early diastolic reverse peak velocity (Vd),systolic diameter (Ds),diastolic diameter (Dd),systolic blood pressure(SBP),dias-tolic blood pressure ( DBP) ,mean arterial pressure ( MAP) and heart rate ( HR) at each time point of before and after anesthesia induction , intubation,cutting the endocranium ,suturing the flaps were observed and recorded .Results Compared with those before anesthesia induc-tion,the MAP,VD and Vs were decreased (P0.05).Conclusion Color doppler ultrasound could reflect the femoral artery hemodynamics of patients who subjected to craniotomy for intracranial hematoma under general anesthesia ,which provides some guid-ance for the general anesthesia strategies .

5.
Article in Chinese | WPRIM | ID: wpr-491934

ABSTRACT

Objective To detect the protective effect of sufentanil preconditioning on hepatic ischemia-reperfusion injury in rats and the role of c-Jun N-terminal kinase signal pathway in the mech-enism.Methods One hundred and sixty-two SD rats(in either gender,weighing 250-300 g)were ran-domly divided into seven groups:Sham-operated group (group S,n = 30 ),ischemia-reperfusion group (group IR,n =30),sufentanil preconditioning group (group SF1:1 μg/kg,n =30;group SF5:5 μg/kg,n =30;group SF10:10 μg/kg,n =30),SP600125 group (group SP,n =30),and dimethyl sulphoxide control group (group DMSO,n =6),different doses of sufentanil was administered 30 min before hepatic ischemia in group SF1,SF5 and SF10.Blood and liver samples were collected from each group at 0(T1 ),1 (T2 ),2 (T3 ),4 (T4 ),and 6 (T5 )hours after reperfusion.Serum alanine amin-otransferase (ALT)and aspartate aminotransferase (AST)were measured by an automatic biochemi-cal analyzer.Malondialdehyde (MDA)and superoxide dismutase (SOD)in liver tissue was measured. Liver sample was stained with HE to observe the hepatic pathological changes.Immunohistochemical method was used to determine the expression of JNK and western blotting was used to detect the ex-pression of P-JNK.Results Compared with group S,levels of AST,ALT increased significantly in group IR,SF1,SF5,SF10 at T1-T5 and in group SP,DMSO at T3 (P <0.05 ).Compared with group IR,levels of AST,ALT decreased significantly in group SF1,SF5,SF10 at T1-T5 and in group SP at T3 (P <0.05).Compared with group S,levels of MDA,SOD increased significantly in group IR,SF1, SF5,SF10 at T1-T5 and in group SP,DMSO at T3 (P < 0.05 ).Compared with group IR,levels of MDA,SOD decreased significantly in group SF1,SF5,SF10 at T1-T5 and in group SP at T3 (P <0.05).Compared with group SF1 and SF5,levels of MDA,SOD decreased significantly in SF10 at T4 . Compared with T1 ,the expression of p-JNK in group IR increased significantly at T3 (P < 0.05 ). Compared with group S,the expression of p-JNK in groups IR,SF1,SF5,SF10,SP,DMSO increased significantly at T3 (P < 0.05 ).Compared with group IR,the expression of p-JNK in groups SF1, SF5,SF10,SP decreased significantly and that in groups SF5,SF10 were less than that in group SF1 (P <0.05 ).The expression of p-JNK in group SF10 was less than that in group SF5 (P < 0.05 ). Conclusion Sufentanil preconditioning can reduce the hepatic ischemia-reperfusion injury and the dos-age of 10 μg/kg was the most effective.The protective mechanisms may inhibit JNK pathway and re-duce the expression of JNK.

6.
Chongqing Medicine ; (36): 3037-3039, 2014.
Article in Chinese | WPRIM | ID: wpr-455960

ABSTRACT

Objective To study the protective effect of sufentanil preconditioning on hepatic ischemia-reperfusion injury in rats and to investigate the mechanisms whether may be by activating p38 MAPK signal pathway to promote p38 MAPK phosphoryla-tion .Methods Thirty SD rats(in either gender ,weighing 220-270 g) were randomly divided into five groups :Sham-operated group (Ⅰ) ,ischemia-reperfusion group(Ⅱ);sufentanil preconditioning group(5 μg/kg ,Ⅲ) ,SB203580(an inhibitor of p38 MAPK) group (Ⅳ) ,and dimethyl sulphoxide (DMSO) control group(Ⅴ) .Sample specimens were collected from each group at 240 minutes after reperfusion .Serum alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were measured by an automatic biochem-ical analyzer .Malondialdehyde(MDA) and superoxide dismutase(SOD) in liver tissue was measured .HE staining was used to ob-serve the hepatic pathological changes ,and to examine the expression of phosphor-p38 mitogen-activated protein kinases (p-p38 MAPK)of hepatic tissues by western blotting .Results Compared with group Ⅰ ,levels of AST ,ALT and MDA showed signifi-cantly increased in group Ⅱ-Ⅴ ,but levels of SOD decreased ,and obvious pathological changes were observed in the liver .In GroupⅢ significantly decreased the elevated levels of ASL ,ALT and MDA but increased levels of SOD ,and lessened hepatic pathological changes ,caused promoted p38 MAPK phosphorylation at 240 minutes after reperfusion .The protective effects of sufentanil precon-ditioning were abolished by SB203580 pretreatment .There were no significant differences between group Ⅴ and group Ⅱ .Conclu-sion Sufentanil preconditioning can reduce the hepatic ischemia-reperfusion injury .The protective mechanisms may be by activating p38 MAPK signal pathway to promote p38 MAPK phosphorylation .

7.
Article in Chinese | WPRIM | ID: wpr-453243

ABSTRACT

Objective To investigate the genetic characteristics of E 1 gene of rubella virus strains isolated in Henan province for further investigation on rubella prevention and control .Methods RNA was extracted from rubella virus strains isolated from suspected measles cases in Henan during 2008 to 2012 .E1 gene of the isolates were amplified by RT-PCR and sequenced .The nucleotide ( nt ) and amino acid ( aa ) sequences of E1 gene of Henan isolates were aligned with the sequences of other reference strains downloaded from GenBank.Phylogenetic tree was constructed by using MEGA 6.0 software package.Results The pre-dominant genotype of rubella virus isolated in Henan was 1E genotype.No 1F genotype was detected.The 2B genotype emerged in 2013.The E1 gene of Henan isolates shared 87.8%-100% homologies in nucleo-tide sequences and 67.0%-100%in amino acids sequences .No variation was detected at hemagglutination inhibition and neutralization sites .A Leu to Phe mutation occurred at amino acid site 338 of 1E genotype in all isolates.Conclusion The results of this study demonstrated that genotype 1E was the predominant geno-type of rubella virus epidemic in Henan province .

8.
Chinese Journal of Trauma ; (12): 1088-1091, 2012.
Article in Chinese | WPRIM | ID: wpr-430746

ABSTRACT

Objective To investigate the risk factors for deep vein thrombosis (DVT) after bone trauma.Methods The study involved 118 patients with traumatic fractures (traumatic fracture group),21 DVT patients diagnosed by color Doppler (DVT group) and 56 healthy patients (control group).Anti-cardiolipin antibody (ACA) was determined by ELISA method.D-dimer and fibrinogen (Fib) were detected by coagulation analyzer and C-reactive protein (CRP) by rate nephelometry.Results Levels of D-dimer,Fib and CRP in traumatic fracture group were significantly increased,but were lower than those in DVT group.ACA positive rate in DVT group presented significant increase and three patients with positive ACA in traumatic fracture group all suffered from DVT.The positive rates of Fib,D-dimer and CRP in lower limb fracture group,multiple fracture group and pelvic fracture group were higher than those in upper limb fracture group (P < 0.05).Levels of Fib and D-dimer showed gradual rise with growth of age,but their levels in DVT group had different degree of reduction after thrombolytic therapy.Conclusions Positive ACA and enhancement of D-dimer,Fib and CRP are risk factors for DVT after bone trauma.Levels of Fib and D-dimer in patients with bone trauma are related with age and therefore risk of posttraumatic DVT increases with age.

9.
Chinese Journal of Dermatology ; (12): 407-410, 2011.
Article in Chinese | WPRIM | ID: wpr-416976

ABSTRACT

Objective To investigate the roles of hemopoietic stem cells (HSCs) in the pathogenesis of psoriasis. Methods HSCs were isolated from the bone marrow of a patient with psoriasis vulgaris and a normal human control. Forward- and reverse-subtracted cDNA libraries were constructed by using suppression subtractive hybridization (SSH) technique between HSCs from the patient and control. Bacterial PCR and dot hybridization were performed to screen for positive clones followed by gene sequencing, identification and functional analysis. Real time quantitative PCR was carried out to measure the mRNA expression of interferon-γ and thymosin 10 (TMSB10). Results Nine genes were screened from the forward-subtracted cDNA library which encoded interferon-γ, tyrosine phosphatase, SUMO1 activase, etc, and 8 genes including the TMSB10-encoding gene were screened from the reverse-subtracted cDNA library. The relative expression level of interferon-γ in HSCs from the patient was 47.5 times that in HSCs from the control, while the level of TMSB10 from the control was 22.6 times that from the patient. Conclusion The abnormal expression of 17 genes which encode interferon-γ, thyrosin, and so on, may be involved in the development of psoriasis.

10.
Chinese Journal of Dermatology ; (12): 441-442, 2011.
Article in Chinese | WPRIM | ID: wpr-416727

ABSTRACT

Objective To explore the significance of FSH1 in the pathogenicity of M. canis. Methods Thirty M. cam's strains from tinea capitis lesions and 30 M. canis strains from tinea corporis lesions were cultured, passaged, and induced by medium containing skin tissue of scalp or foreskin from children. Semi-quantitative reverse transcription (RT)-PCR was carried out to detect the expression of FSH1 mRNA in the firstand fifth-generation M. canis strains, as well as M. canis strains induced by the skin tissues. Results The mRNA expression of FSH1 was higher in M. canis strains derived from tinea capitis lesions than in those from tinea corporis lesions (P 0.05). The skin tissue from scalp and foreskin induced a significant elevation in the mRNA expression of FSH1 in these M. canis strains (F = 2025.713, 1833.139, both P< 0.01), and the inductive effect of the scalp tissue was different from that of the foreskin tissue (P < 0.01). Conclusions The FSH1 mRNA expression is different in M. canis isolated from different body sites. Local skin tissue has an inductive effect on the expression of FSH1 mRNA, and the inductive effect of scalp tissue is more apparent than that of foreskin tissue.

11.
Article in Chinese | WPRIM | ID: wpr-402780

ABSTRACT

Objective To investigate the effect of midazolam as premedication on stress response in patients undergoing cervical plexus block.Methods Sixty female patients undergoing selective thyroid surgery were equally randomized into groups of A and B.The patients in group A were intramuscularly injected midazolam 0.08 mg/kg before cervical plexus block and those in group B were not as the controls.Cervical plexus block was performed with 20 ml mixeture of 1% lidocaine and 0.25% ropivacaine 20 min later.BP and HR were recorded,rate-systolic pressure product (RPP) was calculated,and serum glucose(Glu),cortisol (Cor) and angiotensin Ⅱ (AT-Ⅱ) were detected before enter(T_0),before cervical plexus block(T_1),at 5 min(T_2),15 min(T_3) and 25 min(T_4) after cervical plexus block injection.Results In group B BP,HR,RPP,Glu,Cor and AT-Ⅱ all higher at T_1-T_4 than those at T_0 BP,HR,RPP,Glu,Cor and AT-Ⅱ were all lower in group A than those in group B at T_1-T_4 (P<0.05).Conclusion Intramuscularly premedication with midazolam is effective in reducing stress response in patients undergoing cervical plexus block.

12.
Chinese Journal of Dermatology ; (12): 108-110, 2010.
Article in Chinese | WPRIM | ID: wpr-391259

ABSTRACT

Objective To observe the growth and biological features of bone mesenchymal stem cells (BMSC) from psoriatic patients. Methods Peripheral blood mononuclear cells were isolated from 5 patients with active psoriasis vulgaris and 5 normal human controls, and BMSC were obtained and purified using plastic adherence method followed by primary culture and passage in vitro. The cell morphology, density and growth were observed with microscopy. Cell growth pattern was evaluated by MTT assay. Flow cytometry was applied to identify surface antigens, including CD29, CD34, CD45 and CD106, on these cells. Results No significant difference was observed in the morphology of primary or descendant BMSC between the patients and controls.The primary BMSC from psoriatic patients tended to adhere to the plastic wall later, confluence and grow more slowly compared with those from the controls. The BMSCs from both psoriatic patients anti healthy donors were positive for CD29, but negative for CD34 or CD45. On the 4th day of culture, the BMSC from psoriatic patients exhibited a decrease in proliferation, with the absorbence at 470 nm (A470) being 0.081±0.0066 and 0.095±0.0130, respectively for BMSC from the patients and controls (t=2.358, P<0.05). Conclusion There is a decrease in the proliferation of BMSC from psoriatic patients which show a morphological similarity to those from healthy controls.

13.
Chinese Journal of Dermatology ; (12): 622-624, 2009.
Article in Chinese | WPRIM | ID: wpr-393052

ABSTRACT

ific primer pair S2-R2 can be used to detect S. schenckii in paraffin-embedded tissue.

14.
Chinese Journal of Biotechnology ; (12): 1516-1523, 2009.
Article in Chinese | WPRIM | ID: wpr-296896

ABSTRACT

The heterologously expressed L1 protein of human papilomavirus 16 can assembly into virus-like particles (VLPs), which has been used as prophylactic vaccine for cervical carcinoma. To express L1 protein in Hansenula polymorpha, we analyzed the codon usage of the native gene of L1 protein and redesigned the encoding sequence according to the codon bias of H. polymorpha. We used assembly PCR to synthesize the native gene HPV16L1-N and the codon optimized gene HPV16L1. The synthesized genes were cloned into pMOXZa-A vector to generate plasmids pMOXZ-HPV16N and pMOXZ-HPV16. The expression cassettes MOXp-HPV16L1(N)-AOXTT were cloned into YEp352 vector and transferred into H. polymorpha. After methanol inducement, the expression of L1 protein in H. polymorpha was detected from the codon optimized gene HPV16L1 rather than the native gene HPVI6L1-N. The parameters for induced cultivation for strain HP-U-16L with HPV16L1 were investigated in shaking flask cultures. After induced cultivation in YPM (pH 7.0) medium supplemented with methanol to a final concentration of 1.0% every 12 h at 37 degrees C for 72 h, the recombinant produced 78.6 mg/L of L1 protein. This work offers the possibility for the production of prophylactic vaccine for cervical carcinoma by H. polymorpha.


Subject(s)
Capsid Proteins , Genetics , Cloning, Molecular , Codon , Genetics , Genetic Vectors , Genetics , Human papillomavirus 16 , Genetics , Oncogene Proteins, Viral , Genetics , Pichia , Genetics , Metabolism , Recombinant Proteins , Genetics
15.
Clinical Medicine of China ; (12): 828-830, 2009.
Article in Chinese | WPRIM | ID: wpr-391339

ABSTRACT

Objective To evaluate the clinicophatholgic benefits and safty of antivirus therapy in patients with liver cirrhosis resulting from hepatitis B.Methods 80 patients with HBV-ralated liver cirrhosis were divided into three groups by the histopathology of liver:group of lamivudine treated with lamivudine 100 mg once daily;Adefovir group treated with Adefovir 10 mg once daily;control group treated with liver protective treatment only.Liver and renal function,PTA and HBV DNA were regularly measured.The Child push-Turotte sore and histopathology wag compared before and after treatment.All courses of treatment were 36 weeks.Results The scores of Child Pugh-Turotte sore in groups of lamivudine and Adefovir were lowered sinificantly (3.9 and 2.1 respectively),the load of HBV-DNA was decreased also[(4.1±0.9) copies/ml and(2.8±1.0) copies/ml],liver inflammmation decreased by more than 2 scores and liver fiber was improved by more than one score,with obviously significant difference(P<0.05) as compared with control group.Conclusion Patients with HBV-related cirrhosis treated with lamivudine and adefovir for antivims are improved and antivirus is important and safe to those during cirrhosis decompensation.

16.
Article in Chinese | WPRIM | ID: wpr-525404

ABSTRACT

Objectives To identify genotypes of 31 Sporothrix schenckii (S.schenckii) strains by Southern blotting and to explore the relationship between genotypes and geographic distributions and clinical manifestations.Methods Total DNA was extracted by cetyltriethyl ammonium bromide (CTAB).The polymorphisms were detected by hybridization of ApaⅠ-digested S.schenckii genomic DNA with a probe amplified from the small-subunit rDNA and adjacent internal transcribed spacer (ITS) regions.The band patterns manifested by Southern blotting were employed to investigate genotypes of 31 strains of S.schenckii collected from five different areas in China.Results Of 31 strains of S.schenckii,15 individual patterns (DNA Type A-O) were recognized.Type A to C accounted for 51.61% of all strains.Conclusion The Southern blotting provides a highly sensitive and reliable means for DNA typing of S.schenckii.It is also found that there is an obvious correlation between DNA patterns and different geographic distribution and clinical manifestations.

17.
Article in Chinese | WPRIM | ID: wpr-531403

ABSTRACT

AIM:To investigate ischemia reperfusion (I/R)-induced proteomic changes in rat skeletal muscle. METHODS: Healthy male Wistar rats were randomly divided into two groups as follows (n=6): sham group and I/R group. I/R of right hind limb was induced by 4 h ischemia followed by 24 h reperfusion. The 2-DE was applied to separate the proteins extracted from skeletal muscle tissue at the end of experiment, followed by Coomassie Brillant blue R-250 staining. Computer image analysis was used to determine the differential expression of proteins between the two groups, and 7 protein spots expressed differentially were picked out and subjected to in-gel digest and MALDI-TOP for identification. RESULTS: 354?13 proteins were detected and the match rate was (78.7?1.4)%. 10 proteins displayed significant changes after I/R, of which, 6 proteins increased and 3 proteins decreased in expression. Moreover, 2 spots in I/R group were observed, only 1 spots of which in control. 5 proteins were identified after mass spectrometry. Mitochondrial aldehyde dehydrogenase (ALDH) precursor, heat shock 27 kD protein (HSP27), an unnamed protein product (increased in I/R group), ?-actin (decreased in I/R group), and nuclear transport factor 2 (NTF-2) W7a mutant were found in I/R group. CONCLUSION: I/R injury induced differential proteomic changes in rat skeletal muscle. ALDH, ?-actin and HSP27 expression, and NTF-2 mutation are involved in I/R injury.

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