Objective To explore the relative factors on delayed encephalopathy after acute carbon monoxide poisoning(DEACMP)and the signifiance of P 300 for judging the severity and prognosis of DEACMP.Methods Thirteen aspects such as contact time with carbon monoxide (CO)?coma time et al were analysed between 44 patients with DEACMP (DEACMP group)and 42 patients with acute CO poisoning but without DEACMP (ACMP group). P 300 s were studied in the patients with P 300 as compared with the patients had just caught acute CO poisoning (DEACMP group).and 44 normal controls (NC group).Results There was extremely significant difference in contact time with CO,coma lasting time,treatment time with hyperbaric oxygen,age,complication ( P

Objective To detect the strain features of regional wall abnormalities in patients with coronary artery disease during each diastolic period, and its possibility to evaluate regional left ventricular cardiocyte viability and diastolic function.Methods 54 patients with anterior myocardial infarction (MI group) and 78 normal subjects (NOR group) underwent Doppler tissue imaging, which were performed in 2-chamber-view by strain curves synchronously.Results In the NOR group, strain value of 66 cases (84.62%) showed an gradually increasing negative value from the apex to base to middle of left ventricle, while in the MI group, there were 9 cases (16.67%) with such a trend. IR phase: in the NOR group, 564 segments (90.38%) were upward wave bands, but in the MI group, there were 123 segments (28.47%) having such waves ( P<0.05). RF phase: in the NOR group, 576 segments (92.31%) were upward and steep wave bands, but in the MI group, the number of upward waves were obviously less (102 segments,23.61%) ( P<0.01). SF period: compared with the NOR group, which was horizontal, the MI group had upward wave bands ( P<0.05).Conclusion Regional myocardial ischemia and infarction can cause significant regional diastolic wall abnormalities of strain value in active diastolic phase. Regional diastolic wall motion abnormalities can be evaluated quantitatively and synchronously with high sensitivity by strain curve which has the potential value in cardiocyte viability and diastolic function.

BACKGROUND: Severe acute respiratory syndrome (SARS) is caused by a genetically novel coronavirus that is caused by acute infectious disease. It is not yet clear for the immunology function of SARS patients in their convalescent stage.OBJECTIVE: To study the effects on T lymphocyte, and the titer profiling of 24 T cell receptor (TCR) V β subfamilies expressions in SARS convalescent patients.DESIGN: A self-control observation.SETTING: Central Laboratory, Guangdong Provincial Hospital of Traditional Chinese Medicine.PARTICIPANTS: Seventy-six cured SARS patients who received treatment in the Second Hospital Affiliated to Guangzhou University of Traditional Chinese Medicine between January and April 2003. All the patients corresponded to "clinical diagnostic criteria of atypical pneumonia", " diagnostic criteria of severe atypical pneumonia and discharge criteria" and "clinical diagnostic criteria and discharge criteria of severe acute respiratory syndrome". The involved patients, 30 male and 46 female, averaged (32±11 (years old. Another 10 subjects who simultaneously received health examination in the same hospital, 5 male and 5 female, aged (32±7(years, were involved in the study. Informed consents of detected items were obtained from all the subjects.METHODS：①Detecting the expression of 24 T cell receptor(TCR)V β subfamilies in SARS convalescent patients：Peripheral blood(2 mL) was collected from the healthy convalescent subjects,and EDTA-K2 was used as anticoagulant．In the flow cytometry delection tubes．10 μL various fluorescein-labeled mAb，such as anti-CD3,anti-CD4，anti-CD8,anti-CD25，anti-CD28，anti-HLA-DR，anti-CD3mAb conjugated with PC5，TCR Vβ1(PE+FITC)．Vβ2(PE+FITC)。Vβ3 (FITC)，Vβ4(PE+FITC)，Vβ5．1(PE+FITC),Vβ5．2(PE),Vβ5．3(PE)，Vβ7．1(PE+FITC)，Vβ7．2(FITC),Vβ8(FITC),Vβ9 (PE)，Vβ11(PE)，Vβ12(FITC)，Vβ13．1(PE),Vβ13．2(PE),Vβ13．6(PE+FITC)，Vβ14(FITC),Vβ16(FITC),Vβ17 (PE+FITC)，Vβ18(PE)，Vβ20(FITC)，Vβ21．3(FITC),Vβ22(PE+FITC)and Vβ23(PE)，was added in special flow tubes,and then 50 μL whole blood was added．The mixed solution was incubated away from light for 15 minutes．After erythrocytolysin being added，mixed solution was washed．Finally．cell deposit was dissolved in 300 μl phosphate buffer solution (PBS)．Coulter ESP flow cytometer was used for detection．For the analysis of TCR expression,an electronic gate was set on these cells and at least 5000 events per sample were collected．Three-color cytofluodmetric analysis was performed using a Coulter ESP flow cytometer．②Detecting the T cell subset,activated T and B cells,and the percentage of Ts and Tc cells：5000 cells were collected and used to calculate the expression of T cells (CD3,CD4 and CD8),the activated T and B cells(CD3+／CD25+,CD3+/HLA-DR+ and CD3-／HLA-DR+),as well as the percentage of Ts and Tc cells by Coulter ESP flow cytometer and its software．MAIN OUTCOME MEASURES：①The change of T cell subset(CD3，CD4，and CD8)from SARS convalescent patients．②The change of activated T and B cells(CD3+／CD25+，CD3+／HLA-DR+ and CD3-／HLA-DR+)．③The percentage of Ts and Tc cells(CD8+／CD28+，CD8+/CD28-)in convalescent patients．④Analysis of the 24 TCR V β subfamilies from SARS patients in convalescence．RESULTS：All data were explored to analyze the expression profiling of 24 TCR Vβ subfamilies,the data from 74 SARSpatients and 10 healthy controls were explored to other result analysis．①The detecting results of T celI subset：The percentage of CD4+T cell mean value was lower than the reference value[(33．33±6．64)％ vs．(43±9)％，P＜0．01]．The percentage of CD8+T cell mean value was higher than the refefence value[(34．07±6．40)％ vs．(30±9)％,P＜0．01]．② The expression of activated T and B cells：Percentage of HLA-DR+ T and B cell was Increased while the percentage of CD25+ T-cell was decreased compared with reference values．In 53 out of 74 patients,the percentage of CD25+ T cells was lower than the reference value,and 64 patients had a lower percentage in CD3+/CD25+ T cells．The percentages of CD3+/HLA-DR+ and CD3-／HLA-DR+ cells were higher than the normal reference value．T cells expressing higher CD3+／HLA-DR+ were found in 36 patients，and T cells expressing higher CD3-/HLA-DR+ were found in 30 patients．③The ratios of Ts and Tc cells：The percentage of Ts cells which expressed CD8+／CD28- was increased compared with reference value [(28．75±7．31)％ vs．(15．99±5．1)％，P＜0．01],while the percentage of Tc cells which expressed CD8+／CD28+ was decreased [(5．99±3．60)％ vs．(13．2±4．1)％，P＜0．01]．Thirty-nine patients were found to possess the lower Tc cells and forty-eight patients were found to possess the higher Ts cells．The ratios of both CD4+ and CD8+ T cells were in the normal reference value．④24 TCR Vβ subfamilies expressions in T cells：It was noteworthy that Vβ14 had a highest percentage in all 24 Subfamilies,and followed by Vβ 5．3,and Vβ 23 in the convalescent patients．The percentage of Vβ 14 was the highest in the normal controls,which was consistent with the results of SARS patients．But the other subfamilies expression patterns were different．There were significant differences between Vβ1,Vβ5．2,Vβ5．3,Vβ7．2，Vβ9，Vβ11,Vβ13．1,Vv13．2，Vβ17,Vβ18，Vβ22 and Vβ23．In the convalescent period，each TCR Vβ expression of SARS patients was higher than that of controls(P＜0．05-0．01)．CONCLUSION:In SARS convalescent patients,the increased CD8+CD28- T cell may elevate CD8+ T cell number;Meanwhile.the reduced CD3+ and CD4+ T cell number may be corresponding to the increased Ts cell number．For some inhibiting factor secreted by Ts cell was also increased．The usage pattern of 24 TCR Vβ subfamilies in SARS patients is different from that of control group．The increase of percentage of CD3+／HLA-DR+ and CD3-／HLA-DR+ T cell may be related to the late response of activated T and B cells.

This study examined the effect of P85 (a pluronic block copolymer) and microbubble (MB) ultrasound contrast agents under ultrasound irradiation on gene transfection and expression. The pEGFP plasmids that can encode enhanced green fluorescent protein (pEGFP) served as a report gene and were mixed with different concentrations of MB/0.05% (w/v) P85. Then the plasmids were transfected into human hepatoma G2 (HepG2) cells. The HepG2 cells treated with MB/P85 or without treatment were exposed to ultrasound (US parameters: 1 MHz, 1.0 W/cm(2), 20 s, 20% duty cycle). Twenty-four hours later, the transfection efficiency was assessed by fluorescence microscopy and fluorescence activated cell sorting (FACS) analysis. The cell viability was evaluated by Trypan blue exclusion test. The results showed that the gene transfection efficiency in HepG2 cells under ultrasound irradiation was significantly higher than that without ultrasound irradiation. HepG2 cells in the MB or P85 group in the absence of ultrasound expressed less amount of green fluorescent protein. The expression efficiency reached (22.14±3.06)% and the survival rate was as high as (55.73±3.32)% in the 30% MB plus P85 group. It was concluded that MB and P85 in the presence of ultrasound can enhance gene transfection and expression.

Adult ; Heart Diseases ; etiology ; Hemorrhagic Fever with Renal Syndrome ; complications ; Humans ; Male

One influenza H3N2 virus, A/swine/Shandong/3/2005 (Sw/SD/3/2005), was isolated from pigs with respiratory disease on a farm in eastern China. Genetic analysis revealed that Sw/SD/3/2005 was a triple-reassortant virus with a PB2 gene from human-like HIN1, NS from classical swine H1NI, and the remaining genes from human-like H3N2 virus. These findings further support the concept that swine can serve as reservoir or mixing vessels of influenza virus strains and maintain genetic and antigenic stability of viruses. Furthermore, we have successfully established a reverse genetics system based on eight plasmids and rescued Sw/SD/3/2005 through cell transfection. HI tests and RT-PCR confirmed that the rescued virus maintained the biological properties of the wild type Sw/SD/3/2005. The successful establishment of the reverse genetics system of Sw/SD/3/2005 will enable us to conduct extensive studies of the molecular evolution of H3N2 influenza viruses in swine.

OBJECTIVEThe present study was undertaken to design antisense oligodeoxynucleotides (AS-ODNs) of glial glutamate transporter-la (GLT-1a) and to evaluate the effectiveness of the designed AS-ODNs on the expression of GLT-1a.

METHODSFive sequences of GLT-1a AS-ODNs were designed according to the C terminus specific sequences of GLT-1a mRNA using antisense design software of IDT Com- pany. Western blot analysis was used to evaluate the inhibition effects of the five GLT-1a AS-ODNs on the expression of GLT-la.

RESULTSThe sequence of GLT-1a AS-ODNs with sequence of 5'-GGTTCTTCCTCAACACTGCA-3' could specifically inhibit the expression of GLT-1a in the hippocampal CA1 subfield of rats, while it had no effect on the expression of GLT-1b. This sequence showed similar inhibition on the expression of GLT-la in sham and ceftriaxone (Cef)-treated rats. It could also significantly inhibit the cerebral ischemic preconditioning (CIP)-induced up-regulation in the expression of GLT-1a. The magnitude of the inhibition in sham, Cef- or CIP-treated rats was similar by more than 60%.

CONCLUSIONFrom the designed five sequences of GLT-1a AS-ODNs, we obtained an effective sequence which can specifically inhibit the expression of GLT-1a.

Animals ; CA1 Region, Hippocampal ; metabolism ; Excitatory Amino Acid Transporter 2 ; antagonists & inhibitors ; metabolism ; Ischemic Preconditioning ; Oligonucleotides, Antisense ; genetics ; RNA, Messenger ; Rats ; Up-Regulation

Objective:To further explore the relationship between Type Ⅰ hypersensitivity reaction and the mechanism of the viral hepatitis B through the research of soluble intercellular adhesion molecular-1(sICAM-1) and soluble vascular cell adhesion molecular-1(sVCAM-1) levels in serum in patients with viral hepatitis B related to Type Ⅰ hypersensitivity reaction and the damage of liver cells, and therefore, provide new theory for the perfection of the immunological mechanism of hepatitis B, especially acute hepatitis B.Methods:Serum sICAM-1 and sVCAM-1 levels were measured in 45 patients with viral hepatitis B and 15 normal cases using double antibody sandwich ELISA method. ALT and AST levels were studied using omni automatic biochemistry analyzer and its correlation with sICAM-1 and sVCAM-1 were observed.Results:The sICAM-1 and sVCAM-1 levels of 7 acute hepatitis B were significantly higher than that of the normal cases. ②The sICAM-1 and sVCAM-1 levels of 38 chronic hepatitis B were also significantly higher than that of the normal cases. ③The sICAM-1 and sVCAM-1 levels of 13 cases moderate chronic hepatitis B were significantly higher than that of 16 cases mild chronic hepatitis B. ④The sICAM-1 and sVCAM-1 levels of 9 serious chronic hepatitis B were significantly higher than that of the group of mild chronic hepatitis B. ⑤Levels of sICAM-1 were significantly positively correlated with serum ALT and AST. ⑥sVACM-1 levels were significantly positively correlated with serum ALT and AST. In conclusion, sICAM-1 and sVCAM-1 levels in patients with acute hepatitis B were most remarkably increased, and then serious chronic hepatitis B, moderate chronic hepatitis B, light chronic hepatitis B in sequence.Conclusion:Serum sICAM-1 and sVCAM-1 levels in patients with viral hepatitis B may reflect the damage of liver. ②Examination of serum sICAM-1 and sVCAM-1 levels in patients with hepatitis B can be used to judge the patient’s condition and to diagnose. ③As an important indication of the inflammatory in Type Ⅰ hypersensitivity reaction, sICAM-1 and sVCAM-1 may interfere the appearence of hepatitis B and the procedure of immunological damage of liver cells.

To prepare the RNAse-resistant virus particles containing the partial gene fragments of avian influenza virus H5N1 for use as RNA standard and control in RNA virus detection, the genes coding the coat protein and maturase of E.coli bacteriophage MS2 were amplified by PCR and then cloned into prokaryotic expression vector pET32a to construct the intermediate vector pET32a-MS2. In addition, the gene sequences coding hemagglutinin (HA), neuraminidase(NA) and M protein of the H5N1 virus were also cloned separately to the down-stream of plasmid pET32a-MS2, thus constructing the prokaryotic expression vectors pET32a-NS2-HA, pET32a-MS2-NA and pET32a-MS2-M. These recombinant plasmids were then transformed separately to E.coli BL21(DE3) with induction by IPTG. to express the virus-like particles. The virus-like particles observed under electron microscopy were identified by RT-PCR ,while their stability was confirmed by real-time RT-PCR. In this way, the virus-like particles were successively constructed and identified through PCR amplification, enzymolysis identification and sequencing analysis. These virus-like particles observed under electron microscopy appeared to be circular in shape with a diameter of about 50 nm. Their stability was proved to be rather good. From these observations, it is apparent that these virus-like particles can be used as RNA standard and quality control in the detection of avian influenza virus H5N1.

OBJECTIVETo explore whether fecal calprotectin (f-calprotectin, FC) may be an early marker for the identification of gastrointestinal injury in preterm infants by measuring FC concentration and changes of FC concentration in infants with different perinatal factors.

METHODSFC concentration was measured using ELISA in 76 samples (50-100 mg) obtained from 38 preterm infants (gestation 29 to 33 weeks), at birth and on the third day after birth (the 1st and the 2nd FC levels). The infants were classified into three groups according to the reason for preterm birth: premature rupture of membranes (PROM; n=13), spontaneous preterm birth (SPB; n=5) and indicated preterm birth (IPB; n=20).

RESULTSThere were no significant differences between the 1st and 2nd FC levels in the 38 infants. The 1st FC level in the PROM group was significantly higher than that in the IPB group (P<0.05). The 1st FC level in infants whose mothers received antenatal antibiotics treatment was significantly lower. Infants born by cesarean section had a significantly lower 1st FC level than those born by vaginal delivery (P<0.05). Both the 1st and 2nd FC levels in infants with feeding intolerance were significantly higher than in infants with feeding tolerance (P<0.05). The 2nd FC level was negatively correlated with 1 min Apgar score (r=-0.3, P<0.05).

CONCLUSIONSPremature rupture of membranes and perinatal asphyxia may lead to an increase in the excretion of FC in preterm infants. FC may be used as a marker for early evaluation of gastrointestinal conditions in preterm infants.

Biomarkers ; Feces ; chemistry ; Female ; Humans ; Infant Nutrition Disorders ; metabolism ; Infant, Newborn ; Infant, Premature ; metabolism ; Leukocyte L1 Antigen Complex ; analysis ; Male