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Objective To investigate the clinical significance of plasma neuropeptide Y (NPY)and oxytocin (OXT) and the mRNA level of OXT gene in first-episode schizophrenia (FES) patients and healthy controls (HC).Methods The plasma concentration of NPY and OXT in 55 FES patients and 33 HC were measured by enzyme linked immunosorbent assay (ELISA) and the mRNA level of OXT gene were performed by real-time quantitative polymerase chain reaction (RT-qPCR).Results The NPY levels in FES patients ((1386.88±254.57) pg/ml) were significantly higher than that in HC ((1 140.62±266.63)pg/ml)(P＜0.01),and both the males and females in FES patients ((1 489.97±231.06)pg/ml and (1 279.97±236.30) pg/ml) were higher than their counterparts in HC ((1 305.40 ± 238.80) pg/ml and (965.55 ±165.45) pg/ml) (P＜0.05,P＜0.01).The OXT level in FES patients ((553.26± 180.49) pg/ml) was lower than that in HC ((696.27±280.77) pg/ml) with significant difference (P＜0.05),especially the females FES patients ((597.49±178.63)pg/ml vs (785.51±329.60)pg/ml);but the mRNA expression of OXT gene in FES patients (0.2075 (0.1653,0.3388)) were significantly higher than that in HC (0.1615 (0.1321,0.2200)) (P＜0.05),and there were no differences between the females of the two groups (P＞0.05).Conclusion The FES patients presented abnormal expression of NPY and OXT,and the concentration changes of plasma NPY and OXT will provide referential significance for the diagnosis of schizophrenia(SZ).
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Objective To investigate the clinical significance of plasma neuropeptide Y (NPY)and oxytocin (OXT) and the mRNA level of OXT gene in first-episode schizophrenia (FES) patients and healthy controls (HC).Methods The plasma concentration of NPY and OXT in 55 FES patients and 33 HC were measured by enzyme linked immunosorbent assay (ELISA) and the mRNA level of OXT gene were performed by real-time quantitative polymerase chain reaction (RT-qPCR).Results The NPY levels in FES patients ((1386.88±254.57) pg/ml) were significantly higher than that in HC ((1 140.62±266.63)pg/ml)(P＜0.01),and both the males and females in FES patients ((1 489.97±231.06)pg/ml and (1 279.97±236.30) pg/ml) were higher than their counterparts in HC ((1 305.40 ± 238.80) pg/ml and (965.55 ±165.45) pg/ml) (P＜0.05,P＜0.01).The OXT level in FES patients ((553.26± 180.49) pg/ml) was lower than that in HC ((696.27±280.77) pg/ml) with significant difference (P＜0.05),especially the females FES patients ((597.49±178.63)pg/ml vs (785.51±329.60)pg/ml);but the mRNA expression of OXT gene in FES patients (0.2075 (0.1653,0.3388)) were significantly higher than that in HC (0.1615 (0.1321,0.2200)) (P＜0.05),and there were no differences between the females of the two groups (P＞0.05).Conclusion The FES patients presented abnormal expression of NPY and OXT,and the concentration changes of plasma NPY and OXT will provide referential significance for the diagnosis of schizophrenia(SZ).
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Objective:To construct the lentiviral vector over-expressing Staphylococcus aureus enterotoxin C3 and detect the expression of target gene in vitro.Methods:SEC3 gene were amplificatied by polymerase chain rcaction( PCR).The GV365 lentiviral vectors were digested by AgeⅠenzyme,which was linked to SEC3 gene and then constructed the GV365-SEC3 lentiviral vetor.Positive clones of vectors were identificd by PCR.Then the positive lentiviral vectors were transfected into 293T cells for lentivirus package.The expression of lentiviral vectors was tested by observating cell fluorescence and Western blot.The virus titer was determined by HIV-1 p24 ELISA.Results: SEC3 gene was amplified and successfully bound to the GV365 lentivirus vectors.The sequences of the recombinant plasmid were confirmed correct by PCR and DNA scqucncing.A large mass of green fluorescent cells were observed after transfecting.And the resulting size of 29 kD protein band of protein electrophoresis, which was consistent with the target gene protein.Viral vector titer was 5×108 TU/ml by ELISA detection.Conclusion: Lentiviral vector over-expressing Staphylococcus aureus enterotoxin C3 was successfully constructed,laid the foundation of observing its effect and mechanism against to tumor in vivo and in vitro for later research.
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Objective To investigate the infection situation of Pseudomonas aeruginosa (PA) in patients with the lower respira‐tory tract infection in the department of respiratory intensive care units (RICU ) in recent five years ,and to analyze the changing trend of antibacterial resistence ,in order to guide rational selection of antimicrobial agents .Methods Strains of bacteria were isola‐ted and identified from sputum specimen of patients in the department of RICU and common ward from Jul .2008 to Jul .2013 .The situations of PA infection and antibacterial resistence were analyzed ,and differences of infection rates of PA and antibacterial resis‐tence were compared between RICU and common ward .Results 517 strains of bacteria were detected from sputum specimens of patients in the department of RICU from 2008 to 2013 ,including 141 strains of PA (accounted for 27 .3% ) ,and ticarcilli/clavulanic acid(61 .0% ) was with the highest rate of resistance among 14 drugs and colistin B(9 .2% ) was with the lowest rate of resistance . 378 strains of bacterias were detected from sputum specimens of patients in common wards ,including 125 strians of PA(accounted for 33 .1% ) ,and ticarcilli/clavulanic acid(28 .0% ) was with the highest rates of resistance and colistin B (4 .0% ) was with the low‐est rate of resistance .Conclusion In recent 5 years ,PA might be one of the main pathogenic bacterias of respiratory tract infections in the depatment of RICU in this hospital ,and antibiotic resistance may increse gradually .The antibiotic resistance in the depatment of RICU may be higher than that in cionmon wards .Clinical effective measures should be taken to prevent nosocomial infection ,and rational use of antibiotics should be taken to reduce the emergence of resistant strains .
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Objective To investigate the clinical significance of red blood cell distribution width in the early diagnosis of patients with chronic heart failure.Methods Seventy eight cases of chronic heart failure patients from November 2011 to November 2013 in our hospital were recruited in this study,and were divided into three different heart failure groups (Ⅱ,Ⅲ,and Ⅳ) according to the degree of heart failure; 40 healthy cases were used as control group.The red blood cell distribution width was measured among four groups.The relationship of the red blood cell distribution width and chronic heart failure was analyzed.According to the red blood cell distribution width data,patients were grouped into ＜ 13.1％,13.1％ ～ 14.5％,and ＞ 14.5％ groups.The eventual fatality rates were compared among three groups with different red blood cell distribution width.Results (1) The red blood cell distribution width was (11.63 ±0.71)％,(13.11 ±1.08)％,(14.31 ±1.52)％,and (16.83 ±0.71)％ in the healthy group,andⅡ,Ⅲ,and Ⅳ heart failure groups,respectively.The red blood cell distribution width was gradually increased with the degree of heart failure,with a statistically significant difference among four groups (P ＜ 0.05).(2) The mortality was 0 (0/32),14.8％ (4/29),and 41.2％ (7/ 17) in different red blood cell distribution width groups (＜ 13.1％,13.1％ ～ 14.5％,and ＞ 14.5％),respectively.The mortality was gradually increased with the increased red blood cell distribution width,with a statistically significant difference among three groups (P ＜ 0.05).Conclusions Red blood cell distribution width is of great importance to detect disease degree of chronic heart failure and can guide one to effectively evaluate disease processes and therapeutic effect.
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ObjectiveA HPLC-FLD method was developed to determine the levels of serum AAA in CRIpatients, and to studythe variationof serum AAAinCRI patientsanditsclinical significances. MethodsSerumsampleswerecollected from100healthcontrolsand 80CRI patients. According to 2002 National Kidney Foundation (NKF) staging diagnosis, CRI patients included 4 of stage 2, 12 of stage 3, 12 of stage 4, 52 of stage 5. According to pathogenesis, CRI patients were also divided into 3 groups :chronic nephritis group ( n = 32), DM group ( n = 36), hypertension group ( n = 12 ).Serums were deproteinized by equal volume of 5％ (v/v) PCA and supernate were analyzed direcdy. External standard method was used as quantitative method. The analytical column was Megres C18. 10％ acetonitrile in water was used as mobile phase. Flow rote was 1.0 ml/min. The wavelengths of fluorescence excitation and emission were changed with specific time. The levels of Tyr, Phe and Trp in CRI groups, different CKD stages and different pathogenesis were compared with healthy control groups to evaluated the sensitivity and specificity of serum AAA for CRI diagnosis. ResultsThe linear ranges of the method were 0. 550 -275.000, 3. 050 - 1220. 000 and 0. 049 -49. 000 pμmol/L for Tyr, Phe and Trp, respectively. The limit of detection (LOD) was 0.014 μmol/L for Tyr, 0.500μmol/L for Phe, and 0.005 μmol/L for Trp. The average recovery was 100. 9％, 101.3％ and 98. 5％ for Tyr, Phe and Trp, respectively. Intra-day CVwas 3. 18％ -4. 20％ ( mean was 3. 13％ )and inter-day CV was 3. 18％ -4. 20％ ( mean was 3. 58％ ). The concentration of serum AAA, Tyr and Trp and the ratio of Tyr/Phe in CRI patients were( 135.74 ±23.23 )μmol/L, (52.27 +8.25) μmol/L, (21.49 ±4.25) μmol/L and[0.87(0.68 - 1.05)]μmol/L. which were lower than that in healthy groups (t value was -14. 709, 4.452, 22. 100, U value was 266.000,respectively, P＜0. 05). The concentration of serum AAA, Tyr and Trp and the ratio of Tyr/Phe in healthy groups were ( 174. 47 ± 11.57 ) μmol/L, ( 63.53 ± 4. 68 ) μmol/L, (44. 22 ± 3. 67 ) μmol/L and[0. 97(0. 94 - 1.00)]μmoL/L. There were no statistically significant difference between the different stage of CRI. Compared with the concentration of Tyr, Phe and Trp among chronic nephritis group, DM group,hypertension group, the concentration of Tyr had no significant changes among these three kinds of diseases (P ＞ 0. 05 ). The concentration of Phe had significant changes between Chronic nephritis group and DM group, Chronic nephritis group and hypertension group ( U = 395.00, 114. 00, P ＜ 0. 05 ) ; the concentration of Trp haad significant changes between Chronic nephritis group and DM group ( U = 349.00, P ＜ 0. 05 ).The diagnostic sensitivity and specificity of serum AAA for CRI were 90％ (72/80) and 100. 0％ (100/100).ConclusionsThe method of high-performance liquid chromatography with fluorescence detection ( HPLC-FLD) is simple, rapid, sensitive and specific. Simultaneous determination of serum AAA was benefit to the diagnosis and evaluation of CRI patients.
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OBJECTIVE@#To detect the active efflux gene qac gene in methicillin-resistant Staphylococcus aureus (MRSA) by hem-nested polymerase chain reaction (PCR) and to learn the carrier condition of qac gene.@*METHODS@#The active efflux gene qacA/B and qacB of 80 strains MRSA isolated from clinical specimens from Aug 2006 to March 2008 were amplified in vitro by hem-nested PCR with the primers designed by computers based on qac information of Genbank, and the PCR fragments were sequenced and analyzed.@*RESULTS@#We detected qacA/B in 19 out of the 80 MRSA strains (23.75%) and qacB in 18 out of the 80 MRSA strains (22.5%). Compared with sequences of qacA (NO.X56628) and qacB(NO.AF535087) in the Genbank, 98% and 97% were identical, respectively.@*CONCLUSION@#The active efflux gene qac gene in MRSA is detected by hem-nested PCR. The proportion of qac gene positive strains is high in clinical practice, which is related to its multi-antibiotic resistance.
Sujet(s)
Protéines bactériennes , Génétique , Séquence nucléotidique , Multirésistance aux médicaments , Génétique , Protéines de transport membranaire , Génétique , Staphylococcus aureus résistant à la méticilline , Génétique , Données de séquences moléculaires , Réaction de polymérisation en chaîne , MéthodesRÉSUMÉ
OBJECTIVE To analyze the significance of serum infectious markers in different sources and investigate the prevetion strategy of nosocomial infection in clinical laboratory. METHODS The serum infectious markers(HBsAg,anti-HCV,anti-HIV and syphilis antibody) were detected in outpatient,inpatient and physical check-up people from Jun 2007 to Jun 2008. RESULTS The total percentage of HBsAg,anti-HCV,anti-HIV or syphilis antibody positive in outpatient,inpatient and physical check-up people was 43.30%,13.56% and 7.26%,respectively.The positive rate of HBeAg in outpatient,inpatient and physical check-up people was 12.29%,1.78% and 0.71%,respectively.The average infection rate of syphilis was 1.75% in inpatients,but in inpatients above 60 years old it could be 3.75%.Twenty-one cases were reconfirmed of HIV positive by the reconfirmation laboratory of CDC in Human Province. CONCLUSIONS The positive rates of four serum infectious markers in outpatient or inpatient are markedly higher than that in physical check-up people(? 2=10 117.6,P
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Objective To detect the mecA gene and tst gene of toxic shock syndrome toxin 1 (TSST-1)of Staphylococcus aureus by using PCR and to learn the carrier condition of tst gene.Methods The mecA gene and tst gene of Staphylococcus aureus strains that isolated from clinical sources in our hospital during August 2006 to May 2007 were amplified in vitro using PCR,and to establish the rapid,specific,and sensitive method of detecting tst gene of methicillin resistant Staphylococcus aureus(MRSA).Results The mecA gene and tst gene were detected,and were made the gene sequencing successfully.Forty-one of 84 strains had mecA gene(48.81%),16 of 84 strains had tst gene(19.05%),10 of 84 strains had both of them,and the positive rate was 24.39%(10/41).Conclusion The proportion of tst gene positive strains of MRSA iS high in clinic,and it must be paid more attention.
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Objective A method was developed for the simultaneous determination of tryptophan(Trp) and its metabolites kynurenine(Kyn) and kynurenic acid(Kyna) by high performance liquid chromatography with fluorescence detection(HPLC-FD),and testing serum levels of Trp metabolites in systemic lupus erythematosus(SLE) patients.Methods Serum samples were deproteinized by equal volume of 0.624 mol/L perchloric acid.The analytical column was Hypersil C18 column,and the mobile phase was 0.20 mol/L zinc acetate,8.3 mmol/L acetic acid,and 2.5% acetonitrile;flow rate was 1.5 ml/min.The excitation and emission wave length of fluorescence detector were 365 nm and 480 nm in 0~11 min,344 nm and 404 nm in 11~15.5 min,254 nm and 404 nm in 15.5~20 min,respectively.Results The linear range of Trp was 0.610~196 ?mol/L,the detection limit was 0.005 ?mol/L,and the average recovery was 103.71%.The linear range of Kyn was 0.049~98 ?mol/L,the detection limit 0.025 ?mol/L,and the average recovery was 97.45%.The linear range of Kyna was 1.050~1047 ?mol/L,the detection limit was 0.050 nmol/L,and the average recovery was 100.60%.Inter-and intra-day precisions were both less than 5%.Phenylalanine,tyrosine,and 5-hydroxytryptamine had no interference.The assay was employed to analyze serum samples of SLE patients.The result showed significant difference in Trp,Kyn,and Kyna content,Kyn/Trp ratio between SLE patients and control group.Conclusions A new method was established for simultaneous determination of Trp,Kyn,and Kyna in serum.The method is simple,fast,sensitive,specific,and suitable for applicability to clinical measurement.
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Objective To analyze the relationship between serum Th1/Th2 cytokines levels and autoantibodies against thyroid, and explore the role of Th1/Th2 cellular immunity imbalance in the pathogenesis of autoimmune thyroid diseases(AITD). Methods 21 patients with Graves'desease(GD), 18 cases with Hashimoto's thyroiditis(HT), 17 cases with non-toxic nodular goiter(NTNG) and 20 healthy subjects were enrolled in this study. The serum concentrations of their Th1 cytokines (IFN-?,IL-2) and Th2 cytokines (IL-4,IL-10) were assayed by ELISA. The serum levels of their thyrotropin receptor antibodies(TRAb), thyroglobulin antibodies (TGAb) and thyroid peroxidase antibodies(TPOAb) were measured by routine methods. The relationship between the serum Th1, Th2 cytokines levels and serum TRAb, TGAb, TPOAb levels were analyzed. Results The serum levels of IL-4 and IL-10 in patients with GD were significantly higher than those in patients with HT,NTNG and healthy subjects(P