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Objective To observe the therapeutic efficacy and safety of Chinese herbal fumigation combined with leflunomide (LEF) and prednisone (Pred) in treatment of systemic sclerosis (SSc) complicated pulmonary arterial hypertension (PAH). Methods Totally 99 SSc patients complicated early PAH were randomly assigned to the Western drugs group (WD, 49 cases) and the integrative medicine group (IM, 50 cases). Patients in the WD group took LEF (20 mg) and Pred (15 mg) , once per day. In addition to routine WD program, those in the IM group additionally received Chinese herbal fumigation. All treatment lasted for 6 months. Raynaud's phenomenon (RP) was observed in each group before and after treatment. RP score, erythrocyte sedimentation rate (ESR), C reactive protein (CRP), pulmonary arterial systolic pressure (PASP) , and pulmonary function were compared between the two groups before and after treatment. The clinical efficacy and adverse reactions were evaluated. Results Thirteen cases were lost due to various reasons. A total of 86 patients completed this study, 41 in the WD group and 45 in the IM group. Compared with the same group before treatment, RP score, levels of ESR and CRP all decreased in the two groups after treatment (P <0. 05). Compared with the WM group after treatment, RP score, levels of ESR and CRP were obviously lowered in the IM group after treatment (P < 0. 05). Besides, lowered differences between post-pre-values of ESR, CRP, and PASP were more obviously higher, while elevated differences between post-pre-values of total lung capacity (TLC) and carbon monoxide diffusing capacity (DLCO) were more obviously higher in the IM group (P <0. 05). The total effective rate was 93. 33% (42/45) in the IM group, obviously higher than that in the WD group [70. 73% (29/41) , P <0. 05 ]. There was no statistical difference in total adverse reaction rate between the two groups (x² =0. 019, P =0. 891). Conclusion Chinese herbal fumigation combined with WD had obvious efficacy with less adverse reactions, so it was worth clinical spread.

Objective: To investigate the differences of clinical features, pulmonary function tests, chest imaging and prognosis between patients with idiopathic pulmonary fibrosis (IPF) combined with emphysema and without emphysema., Methods: Of the 117 patients diagnosed as IPF for the first time in the First Affiliated Hospital of China Medical University from January 2002 to January 2007, 23 cases met the diagnostic criteria for combined pulmonary fibrosis and emphysema (CPFE). Thirty-three cases with isolated IPF from 2002 to 2003 were selected as the control group. Clinical characteristics, pulmonary function indexes, PaO₂, bronchoalveolar lavage fluid (BALF) results and survival time were retrospectively analyzed and compared between the 2 groups., Results: The smoking indexes (pack-years)were significantly higher in the CPFE group (28 ± 18) compared with the IPF group (18 ± 16), t = 2.10, P < 0.05. No significant difference was observed between the 2 groups in terms of age, gender and the percentage of smokers. The CPFE group showed significantly lower FEV₁/FVC% [(77 ± 7)%, (83 ± 8)%, t = 3.55, P < 0.05] and D(L)CO% pred [(44 ± 12)%, (54 ± 16)%, t = 2.48, P < 0.05] compared to those of the IPF group, while TLC%pred was significantly higher in the CPFE group [(77 ± 11)%, (64 ± 12)%, t = 3.93, P < 0.05]. VC% pred and PaO₂ showed no significant difference between the 2 groups. The total cell count, the percentages of macrophages, neutrophils, lymphocytes and eosinophils in BALF showed no significant difference between the 2 groups. There was no significant difference in median survival time between the 2 groups., Conclusion: Smoking was an important risk factor for IPF combined with emphysema. The pulmonary function of IPF combined with emphysema is characterized by a mild abnormality in the lung volume but a significant decrease in the diffusing capacity. Combined emphysema has no influence on the survival time of patients with IPF.

The receptor-binding domain(RBD) protein of HCoV-NL63 is a major target in the development of diagnostic assay and vaccine, it has a pivotal role in receptor attachment, viral entry and membrane fusion. In this study, we prepared 2 purified recombinant HCoV-NL63 RBD proteins using in E. coli system and identified the proteins by Western blotting. We first optimized codon and synthesized the RL (232-684aa)coding gene, then amplified the RL or RS(476-616aa) coding gene via PCR using different primers . The RL or RS coding gene was cloned into the pM48 expression vector fused with TrxA tag. The RBD (RL and RS) of HCoV-NL63 were expressed majorly as inclusion body when expressed in E. coli BL21pLys S under different conditions. The expressed products were purified by affinity chromatography then analyzed by SDS-PAGE and Western blotting. Our results showed that the recombinant RBD proteins were maximally expressed at 37 degrees C with 0. 8mM IPTG induction for 4h. RL or RS protein with 95 % purity was obtained and reacted positively with anti-sera from mice immunized with the recombinant vaccinia virus (Tiantan strain) in which HCoV-NL63 RL or RS protein was expressed. In conclusion, the purified recombinant RBD proteins(RL and RS)derived from E. coli were first prepared in China and they might provide a basis for further exploring biological role and vaccine development of HCoV-NL63.

The aim of the present study was to investigate the effect of the nuclear factor‑κB (NF‑κB) inhibitor, pyrrolidine dithiocarbamate (PDTC), on high‑mobility group protein B1 (HMGB1) expression in a rat model of chronic obstructive pulmonary disease (COPD). The COPD model was developed by administering lipopolysaccharides to the airways of rats and smudging (group B). In addition, a model of COPD complicated with hypoxia was established by administering lipopolysaccharides to the airways of rats, smudging and hypoxia (group C). PDTC was administered to the treatment groups by intraperitoneal injection. Reverse transcription polymerase chain reaction (RT‑PCR) and western blot analysis were used to detect the expression of HMGB1 and NF‑κB in lung tissue. RT‑PCR and western blot analysis demonstrated that HMGB1 mRNA and protein expression in groups B and C increased significantly (P<0.05) compared with control group A. In addition, HMGB1 expression in groups B and C gradually increased. HMGB1 mRNA and protein expression in groups B1 and C1 decreased (P<0.05) compared with B and C. NF‑κB protein expression in groups B and C increased significantly (P<0.05) compared with A. NF‑κB protein expression in groups B1 and C1 decreased compared with B and C. Therefore, HMGB1 mRNA and protein expression was identified to be positively correlated with NF‑κB protein expression. The NF‑κB inhibitor, PDTC, was demonstrated to significantly inhibit HMGB1 expression in lung tissues of rats with COPD and this mechanism may be associated with the NF‑κB signal transduction pathway.

Objective: To establish the fingerprint of Polygonum multiflorum from Guizhou and provide a standard for its quality control., Methods: HPLC analysis was performed on Agillent ZABAX-C18 (4.6 mm x 250 mm, 5 microm), gradient eluted composed of acetonitrile-0.4% water solution of phosphoric acid. Column temperature was set at 25 degrees C and the flow rate was 1 mL/min. The detection wavelength was 280 nm and the analysis time was 60 min., Results: 9 common peaks were identified. The RSD of the relative retention time and the relative peak area were less than 3% in analyzing its precision, stability and repeatability of the common peaks, and the similarity of the 16 batches of sample was more than 0.9., Conclusion: The method is simple and reliable, and it can provide a standard and guidance for quality control of Polygonum multiflorum.

Background: Timely diagnosis and prognostic assessment of ventilator-associated pneumonia remain major challenges in critical care., Objective: To explore the value of soluble triggering receptor expressed on myeloid cells 1, procalcitonin, and the Clinical Pulmonary Infection Score in the diagnosis and prognostic assessment of ventilator-associated pneumonia., Methods: For 92 patients, bronchoalveolar lavage fluid was cultured for detection of microorganisms, serum levels of the receptor and procalcitonin and levels of the receptor in exhaled ventilator condensate were measured, and the Clinical Pulmonary Infection Score was calculated., Results: On the day of diagnosis, patients who had pneumonia had higher serum levels of the receptor, procalcitonin, and C-reactive protein; higher white blood cell counts; and higher pulmonary infection and Sequential Organ Failure Assessment scores than did patients without pneumonia. White blood cell count (odds ratio, 1.118; 95% CI, 1.139-1.204) and serum levels of the receptor (odds ratio, 1.002; 95% CI, 1.000-1.005) may be risk factors for VAP. Serum levels of the receptor plus the pulmonary infection score were the most reliable for diagnosis; the area under the receiver operating characteristic curve was 0.972 (95% CI, 0.945-0.999), sensitivity was 0.875, and specificity was 0.95. For 28-day survival, procalcitonin level combined with pulmonary infection score was the most reliable for prognostic assessment (area under the curve, 0.848; 95% CI, 0.672-1.025)., Conclusions: In patients with ventilator-associated pneumonia, serum levels of the receptor plus the pulmonary infection score are useful for diagnosis, and procalcitonin levels plus the pulmonary infection score are useful for prognostic assessment.

Objective: To explore the diagnostic and prognostic roles of serum total RNA in patients with sepsis., Methods: Seventy-six patients with sepsis admitted to intensive care unit (ICU) of Chinese PLA General Hospital from October 2009 to November 2010 were enrolled. Blood samples were drawn from these patients and total RNAs of these blood samples were isolated. The concentration of total RNA in the samples was detected. C-reactive protein (CRP) was determined by scattering turbidimetry, and procalcitonin (PCT) was determined using enzyme-linked fluorescence analysis (ELFA). Acute physiology and chronic health evaluation II (APACHE II ) score and sequential organ failure assessment (SOFA) score were recorded. Twenty-four healthy individuals with matched age and sex were enrolled as normal controls. Receiver operating characteristic curve (ROC curve) was used to determine the optimal concentration of serum total RNA for the diagnosis of sepsis. Binary logistic regression analysis was used to analyze the risk factors of sepsis patients., Results: The concentration of serum total RNA (ng/μl) in sepsis group was significantly higher than healthy controls [4.233 (0.204, 37.733) vs. 1.593 (0.319, 5.913), P<0.01]. The area under curve (AUC) of the diagnosis of sepsis was 0.820. When the cut-off point was set at 2.027 ng/μl, the concentration of serum total RNA yielded a sensitivity of 95.9% and a specificity of 65.2%. Total RNA concentration in the healthy controls, the survival group (n=42) and non-survival group (n=34) were step wisely increased [1.593 (0.319, 5.913), 4.178 (0.204, 10.776), 5.006 (0.997, 37.733), respectively, F=8.061, P=0.001]. Binary multivariate logistic regression analysis showed that APACHE II score and SOFA score were independent risk factors of septic patients [APACHE II score: odds ratio (OR)=1.265, 95% confidence interval (95%CI) 1.031 to 1.553, P=0.025; SOFA score: OR=1.151, 95%CI 1.031 to 2.284, P=0.012]. But PCT was not independent risk factor of sepsis patients (OR=1.075, 95%CI 0.974 to 1.187, P=0.151). In the binary univariate logistic regression analysis model, the OR of total RNAs was 1.149, but P value was 0.061, because of the small sample size., Conclusions: The concentration of total RNA in sepsis patients' serum was significantly increased and correlated to the severity of sepsis, which can be used as biomarkers for sepsis patients.

Objective: To determine the antigen characteristics of different fragments of SARS-CoV N protein expressed in E. Coli and their application in the serological diagnosis., Methods: Based on preliminary analysis of 39 different segments of the N protein, We choosed six purified N protein for further antigenicity characterization in this study, including that PN360 (1 -360aa), PN301 (1-301aa), PN199 (30-228aa), PN185 (30-214aa), PN155b (60-214aa), and PN125 (90-214aa). We developed Western-Bolt and ELISA to detect antibody reactivity between truncated N fragments with sera from SARS-CoV-negative normal adults or SARS-CoV patient convalescent sera., Results: Western-Bolt results show that all the six fragments have reacted with the SARS patient convalescent sera, but the PN360 and PN301 showed obvious cross-reaction with sera from SARS-CoV-negative normal adults; sensitivity analysis using an ELISA coating with PN199, PN185, PN155b, PN125 as antigen showed that the PN185 and PN155b are better than PN125., Conclusion: Truncated N protein PN185 and PN155b expressed in E. Coli are better antigen candidates used for detection of SARS-CoV specific antibody.

The title compound, C(11)H(12)N(2)O(2), shows an S configuration, in which the pyrrolidinone ring is twisted with respect to the phenyl plane, making a dihedral angle of 70.73 (7)°. In the crystal, mol-ecules are linked by N-H⋯O hydrogen bonds, building up a layer parallel to (001).

Objective: To investigate the impact of prior cerebral infarction (PCI) on in-hospital mortality in patients with Acute Myocardial Infarction (AMI)., Methods: A retrospective analysis of documents of a total of 3572 consecutive patients with AMI admitted to Xuanwu Hospital of Capital Medical University from 2002 Jan. 1 to 2009 Dec. 31 were performed., Results: There were 564 patients (15.8%) with PCI. Compared with the group of without PCI, the group with PCI were substantially older [(69.4 ± 9.9) vs (64.2 ± 12.9) years, P = 0.000], and had a higher prevalence of hypertensive disease, diabetes mellitus, prior myocardial infarction (MI) and non-ST-segment elevation myocardial infarction (NSTEMI) (respectively, 71.0% vs 57.3%; 41.0% vs 25.7%, 12.9% vs 9.5%; 14.9% vs 10.7%, P < 0.01), and a higher in-hospital mortality (16.5% vs 10.0%, P = 0.000). Univariate analysis demonstrated that in-hospital mortality associated with age, gender, extensive anterior MI, anterior MI, diabetes mellitus, prior cerebral infarction, prior myocardial infarction, coronary angiography and percutaneous coronary intervention. Logistic regression analysis found that risk factors were age, extensive anterior MI, anterior MI, diabetes mellitus and prior cerebral infarction, and protective factors were coronary angiography and percutaneous coronary intervention. PCI was independently associated with in-hospital mortality, OR 1.368, 95%CI 1.047 - 1.787, P = 0.022., Conclusion: In patients with acute myocardial infarction, the presence of PCI increases the risk of worse in-hospital outcome.

Objective: To evaluate the value of non-invasive positive pressure ventilation (NPPV) in immunocompromised host (ICH) complicated by acute respiratory failure (ARF), and to investigate predictive variables of success with NPPV in ICH with ARF., Methods: A retrospective study of immunocompromised patients with ARF, who were admitted to respiratory intensive care unit (RICU) from March 2008 to March 2011, was performed. Based on clinical data, univariate Logistic regression was done for prediction for independent factors affecting the success of NPPV treatment. Immunization status was assessed according to clinical outcome., Results: NPPV was instituted in all 33 cases with ARF initially. Among these patients, 9 patients (27.3%) received sequential invasive mechanical ventilation (IMV, failure group) and all of them died finally; among 24 cases (72.7%) who only received NPPV (success group), 7 patients died (29.2%). There was significant difference between the two groups in mortality (P<0.01). The simplified acute physiology scoreII (SAPSII) in the success group was lower than that in the failure group (33±9 vs. 43±5, P<0.01). However, other clinical data showed no statistical significance between two groups. Univariate Logistic regression analysis identified SAPSII was the independent factor associated with the success of NPPV treatment [odds ratio (OR) =0.83, 95% confidence interval (95% CI) 0.709-0.964, P<0.05]. And SAPSII≥38 was a risk factor for the failure of NPPV [area under receiver operating characteristic (ROC) curve 0.73]. In addition, the lung injury scores (LIS) in the survival group was significantly lower than that of the death group (1.95±0.48 vs. 2.57±0.52, P<0.01), the difference was statistically significant. CD3(+) and CD8(+) T counts in the survivors were higher than that of non-survivors (CD3(+):0.73±0.16 vs. 0.41±0.20; CD8(+): 0.51±0.18 vs. 0.21±0.15, both P<0.01), and the difference was statistically significant., Conclusion: As an early treatment for ICH with pulmonary infections suffering from ARF, NPPV can be effective for the ICH patients suffering from severe pulmonary infection through improving hypoxemia, ameliorating respiratory distress symptoms, and avoiding complications associated with IMV when SAPSIIis less than 38. CD3(+), CD8(+), and the LIS can be used to evaluate the prognosis of those patients.

The spike (S) glycoprotein of HCoV-NL63 is a major target in the development of diagnostic assays and vaccines, but its antigenic and immunogenic properties remain unclear. Four fragments coding spike proteins (S1, S2, RL and RS) from HCoV-NL63 were amplified and cloned into the expression vector derived from vaccinia virus (Tiantan strain), and recombinant vaccinia viruses expressing four segments of spike proteins were generated (vJSC1175-S1; vJSC1175-S2; vJSC1175-RL; vJSC1175-RS), respectively. Their expression location in cell and level were characterized using indirect immune fluorescence assay (IFA) and Western-Blot, respectively. The expressions of four segments of spike proteins in recombinant vaccinia viruses were showed at appropriate level and with posttranslational modification (glycosylation), and S1, RL and RS were mainly distributed in the cell membrane, while the S2 was mainly distributed in the cytoplasm. Our results provide a basis for further exploring diagnostic role and vaccine development of different spike segments from HCoV-NL63.

Based on the synergistic effect of pulsed discharge plasma and titanium dioxide (TiO2) photocatalysis, a coupling system of pulsed discharge plasma/TiO2 was set up in the present paper, by using the ultraviolet radiation produced during the pulsed discharge process. TiO2 film immobilized on the glass beads was used as the photocatalyst in the synergistic system. In the research, the synergistic mechanism of pulsed discharge plasma and TiO2 photocatalysis was reviewed by the spectral diagnosis of oxygen radical (* O radical) produced in different reaction systems. The obtained results showed that the characteristic emission spectrum of * O radical could be observed at 777 nm and the corresponding transitions was 3p5 P --> 3s5 S. When air was used as bubbling gas, the relative emission intensity of * O radical recorded in the synergistic system was stronger than that in the plasma alone system, and the same result was also obtained when oxygen (O2) was used as bubbling gas. This result revealed that the photocatalytic activity of the immobilized TiO2 in the synergistic system could be induced by the ultraviolet radiation formed during the pulsed discharge process. In the systems of pulsed discharge plasma alone and pulsed discharge plasma combined with TiO2 photocatalysis, the relative emission intensity of * O radical in distilled water was stronger than that in phenol solutions. The results showed that * O radical was one of responsible radicals for phenol degradation in the pulsed discharge plasma alone system as well as the synergistic system.

Objective: To investigate the role of homeobox gene Nkx2-5 in cardiac myogenesis., Methods: Two P19 cell lines, namely cells transfected with exogenous expression of Nkx2-5 gene and non-transfected cells, were cultured in suspension for 4 days to induce cell aggregation, and the cell aggregates were transferred to the Petri dish for further adherent culture. On days 4, 8, 12 and 16 of adherent culture, the expressions of α-sarcomeric actin (α-SA) and cardiac troponin T (cTnT) protein were detected by immunocytochemistry, and the mRNA expressions of GATA-4, α-myosin heavy chain (α-MHC) and atrial natriuretic factor (ANF) genes by RT-PCR., Results: In the transfected cells, α-SA and cTnT protein expressions were detected on days 8, 12 and 16 of adhere culture, and their expressions increased gradually with time. α-SA and cTnT expression was significantly higher on day 16 than on day 8 of culture (P<0.01). RT-PCR analysis of the transfected cell showed the presence of GATA-4 expression on day 4 of adherent culture, and the expression increased on days 8 and 12 but decreased on day 16. ANF and α-MHC expressions were found on days 8, 12, and 16, increasing gradually over time and showing significant differences from those on day 4 (P<0.05 or P<0.01). The expression of α-MHC was significantly higher on days 12 and 16 than on day 8 (P<0.05 or P<0.01), and ANF expression was significantly higher on day 16 than on days 8 and 12 (P<0.01). The non-transfected cells were negative for the expressions of all these genes., Conclusion: Exogenous expression of Nkx2-5 gene can induce P19 cells to express cardiac markers in vitro.

Background: Recurrence of hallux valgus is considered to be the most common problem experienced postoperatively. We designed and carried out operations to correct hallux valgus by transferring the extensor hallucis longus (EHL) tendon to reduce the likelihood of recurrence., Methods: Twenty-five patients (38 feet) with the average age of (46.3 ± 12.3) (range, 22 to 60) years underwent the operation. The American Orthopaedic Foot and Ankle Society score and weight-bearing radiographs of the foot were applied to assess the feet pre- and postoperatively with a mean duration of follow-up of (38.2 ± 3.2) months. The surgical procedure includes the release of the distal soft tissues, excision of the medial eminence, plication of the medial part of the capsule, and transfer of the EHL tendon, and reconstructing its insertion., Results: At follow-up, 35 feet (23 patients, 85%) were free from pain at the first metatarsophalangeal (MTP) joint. In three feet (two patients), the pain was alleviated but persisted. The mean hallux valgus angle decreased significantly from a preoperative 38.3° ± 8.0° to 7.3° ± 2.0° at the time of the most recent follow-up (P < 0.0001), and the mean intermetatarsal (IM) angle decreased significantly from preoperative 12.5° ± 3.4° to postoperative 6.5° ± 2.4° (P < 0.0001). The mean score according to the American Orthopaedic Foot and Ankle Society had increased from 46.5 to 84.8 points (P < 0.0001)., Conclusions: Hallux valgus can be corrected by transferring the EHL tendon medially and reconstructing its insertion. The technique can achieve stress balance of metatarsophalangeal joints and therefore prevent the recurrence of hallux valgus.

Objective: To express the nuclear capsid protein (N protein) and the spike protein (S protein) of HCoV-HKU1, and to develop the corresponding serum assay for antibody detection., Methods: The N protein of HCoV-HKU1 was expressed in E. Coli, anti-N antibody assay was established using Western Blotting with turn-based membrane. HCoV-HKU1 S protein was constructed in the eukaryotic expression plasmids, and confirmed by Western Blotting, S antibody assay was established using indirect immunofluorescence assay (IFA). We analyzed anti-S and anti-N antibody among 100 normal adult serum., Results: Expression of S and N protein were confirmed; 100 normal adult serum were analyzed using the established serological detection assay, in which HCoV-HKU1 S antibody positive rate was 47%, N antibody positive rate was 48%, Both S and N antibodies positive were 21%, Both S and N antibodies negative were 22%. Co-detection S and N antibody was achieved 74% positive rate., Conclusion: The methods we established here could be used for serological analysis of HCoV-HKU1. Either detection of HCoV-HKU1 S or N antibodies achieved good results. Higher positive detection rate of anti-S or anti-N antibody was found in the normal adults.

To study IgG antibody persistence and temporal change in SARS coronavirus (SARS-CoV) infected patients, 22 patients recovered from SARS in Beijing were recruited and followed-up from 2004 to 2008, serum samples from patients were collected every year. We checked and analyzed the SARS-CoV IgG antibody (Ab) for five consecutive years using the commercial ELISA test kit. The results showed that: all of the serum were SARS-IgG antibody-positive the first year after recovery, the titer of most serum remained at high levels at the 2ed and 3rd year post infection. The Ab titers significantly declined at 4th year post infection. The IgG Ab was almost undetectable after 5 years post infection. In conclusion, SARS-CoV IgG Ab can be maintained for more than 3 years post infection, however, the titer of IgG Ab has declined markedly 4 years later. These data provide a useful reference for diagnosis and control of SARS infection, the evaluation of immune response and vaccine efficacy.

To understand the effect of various gene structures of HIV B'/C subtype on the gene expression and immunity in DNA vaccine, replicating DNA vector pSCK2 was used to construct seven DNA vaccines carrying one or more of HIV B'/C subtype genes: gagpol, gp160 and rtn (rev, tat and nef fusion gene). Immunofluorescence staining indicated that Gag, Gp160, Rev, Tat and Nef could be expressed from the seven DNA vaccines. Stronger expression was observed with the gene in single-gene expression plasmid or with the gene located at upper-IRES in double- or multi-gene expression plasmid. ELISA test showed that Gag induced higher antibody response, but the antibody titers stimulated by Gp160, Pol, or RTN were very low. Both Gag single-gene expression plasmid and Gag-RTN double-gene expression plasmid separately inoculating induced stronger antibody response against Gag than Gag-Gp160 double-gene expression plasmid and Gagpol-Gp160-RTN multi-gene expression plasmid or combined inoculation of Gag and Gp160 single-gene expression plasmids did. ELISPOT detection showed that all the seven DNA vaccines could stimulate cellular immune response against Gag, Pol, Gp160, Tat, and Nef, respectively. Gagpol or Gp160 single-gene expression plasmid separately inoculating stimulated the strongest cellular immune response. Tat and Nef expressed in all the plasmids induced similar immune response. These results indicated that HIV B'/C subtype genes gagpol, gp160 and rtn could be efficiently expressed in the replicating DNA vaccine vector, single-gene expression plasmid had the higher gene expression level and induced stronger immune response; combined immunization of Gagpol and Gp160 had dramatically lower immunity than Gagpol or Gp160 separated immunization did. Immunity of RTN had no difference between combined and separated immunizations. Therefore, in case of immunization with DNA vaccines containing different HIV genes, it is necessary to optimize the combined immunization procedure, especially for the combination of Gag and Gp160-containing vaccines.

Objective: To investigate the effect of vitamin E on renal ischemia/reperfusion injury (RI/RI) in young and aged rats., Methods: The model of (RI/RI) was induced by bilateral clamping the renal artery and vein for 45 min followed by reperfusion. The contents of BUN, Scr, MDA, SOD, NO and iNOS were measured. Proteins of HSP70 were observed by immunohistochemistry. Flow cytometer was used to estimate the apoptosis rate of renal cortex cells., Results: After ischemia/reperfusion injury, the contents of BUN, Scr and iNOS were increased. Compared with young ischemia/reperfusion group, the contents of MDA were higher and the contents of SOD were lower in aged ischemia/reperfusion group. The expression of HSP70, the contents of NO and the apoptosis rate of renal cortex cells were higher in aged ischemia/reperfusion group than that in control group. Vitamin E significantly decreased the contents of BUN, Scr, MDA and iNOS, and increased the contents of NO and SOD after RI/RI. The expression of HSP70 was increased in both VE groups than that in both I/R groups. The apoptosis rate of renal cortex cells was less in both VE groups than that in both I/R groups., Conclusion: Vitamins E may up-regulate the expression of HSP70, increase the contents of SOD and NO, and enhance an ability of clear free radicals, which may contributes to decreasing renal ischemia/reperfusion injury in young and aged rats.

Objective: To investigate the effects of clobenpropit and histidine on reinstatement of morphine-induced conditioned place preference (CPP) in rats., Methods: The persistence, extinction and reinstatement of morphine-induced CPP were established.In clobenpropit group three different doses of clobenpropit (2, 5 and 10 microg/rat, i.c.v.) were administered 15 min after morphine (1 mg/kg, i.p.) was injected. In histidine group histidine (100, 200, 500 mg/kg) was given 1 h prior to morphine treatment (1 mg/kg i.p)., Result: The CPP was reinstated by priming injection of 1 mg/kg morphine. Clobenpropit (5, 10 microg/rat) significantly inhabited the reinstatement by a priming dose of morphine-induced CPP compared with the morphine control group; histidine (100, 200, 500 mg/kg) significantly inhibited the reinstatement in a dose-dependent manner., Conclusion: Clobenpropit and histidine inhibit the revival of morphine-induced CPP in a dose dependent manner, indicating that endogenous histamine may inhibit relapse of morphine to some extent.

Objective: To evaluate the influence of Chinese drugs for cooling blood and dissolving stasis (CBDS) in different concentrations on morphology of krypton laser induced choroidal neovascularization (CNV) in brown Norway (BN) rats., Methods: Forty-eight rats received laser irradiation (659 nm) on fundus of one eye (power 360 mW, spot diameter 50 microm, time 0.05 s). They were divided into four groups equally: the control group (A) treated with normal saline, and the three CBDS groups treated respectively with high (B, 5.0 g/kg), median (C, 2.5 g/kg) and low (D, 1.25 g/kg) dosage of CBDS, twice every day via gastric perfusion for 21 successive days. Fundus fluorescein angiography (FFA) on 4 selected rats in each group was performed at the 7th, 14th and 21st day after photocoagulation, and histopathologic examination using light microscope with immuno-histochemical stain was conducted on them as well., Results: FFA showed that CNV was firstly appeared on day 7 after photocoagulation, in Group A, it expanded gradually and reached the peak on day 21, but showed no significant expansion in the three CBDS groups. The fluorescein leakage in Group C (52343.13 +/- 12973.92 dots) and D (66252.78 +/- 20659.71 dots) was significantly less than that in Group A (91457.19 +/- 29309.11 dots) and B (95973.40 +/- 53950.43 dots) on day 21, all showing statistical significance (P<0.05). The variation of CNV in thickness showed that in Group A it increased gradually from day 7 and reached the peak on day 21 (55.3383 +/- 8.5036 microm); but in the CBDS groups, the peak was reached on day 14, then became thinned gradually, on day 21, it was 40.0913 +/- 13.3448 microm in Group B, 38.8473 +/- 7.9862 microm in Group C and 38.9372 +/- 5.1728 microm in Group D, all thinner than that in Group A significantly (P<0.05)., Conclusion: CDBS can effectively suppress the krypton laser induced CNV proliferation and prevent the CNV leakage in BN rats.

HA and M2 genes derived from human highly pathogenic avian influenza H5N1 virus (A/Anhui/ 1/2005) isolated from China, were amplified and cloned into the DNA vaccine expression vector pVRC. In order to improve the expression of hemagglutinin, the human codon usage preference was made and the whole length of HA gene of H5NI (A/Anhui/1/2005) influenza virus was synthesized,named HA/YH/K, and inserted to pVRC vector, the expression of HA/YH/K protein in eukaryotic cells was significantly improved according to internal control of actin protein. Furthermore, the M2 and HA/YH/K genes were cloned into bicistronic eukaryotic expressing vector pIRES to yield the recombinant plasmid pIRES-HA/ YH/K-M2/YS/K, which could expressed HA and M2 protein simultaneously by transfection of one plasmid. Western blot and IFA showed that the recombinant pIRES-HA/YH/K-M2/YS/K plasmid was successfully expressed in several mammalian cells (Hela, MDCK and 293FT). The above results may help to identify the function and pathogenic mechanism of HA, M2 genes derived from HPAI H5N1 (Anhui strain) and pave a way for the development of novel bivalent vaccines against human highly pathogenic avian influenza virus and for preparedness for influenza pandemic.

NOD/SCID (non-obese diabetic/severe combined immunodeficient) mice are immune deficient mice which are made by backcross of severe combined immunodeficient mice with non-obese diabetic mice strains. NOD/SCID mice are both innate immune deficiencies and lack of T and B lymphocytes. Various tumor cells can be implanted in this kind of mice, the rejection and graft-versus-host disease (GVHD) occur fewer. Therefore, NOD/SCID mice gradually become a useful tool for the study on Experimental Hematology. This paper comprehensively reviews the biological characteristics of NOD/SCID mice, the establishment of human leukemia model, stem cell transplantation, drug research, deficiency and improvement of NOD/SCID mice in application for study.

In the title compound, [Pr(C(32)H(30)N(2)O(6))(NO(3))]·CHCl(3), the Pr(III) ion is ten-coordinated by eight O atoms and two N atoms from the acyclic crown-type Schiff base ligand and the bidentate nitrate group. The coordination polyhedron around Pr(III) is a distorted bicapped square anti-prism. The chloro-form solvent mol-ecule is not involved either in coordination to the Pr(III) center or in hydrogen bonding to the complex. The Pr-O(phenolate) bonds are significantly shorter than the Pr-O(ether) and Pr-O(nitrate) bonds, which suggests that the Pr-O(phenolate) bond is stronger than these other bonds. In the crystal structure, the acyclic crown-type Schiff base ligand wraps around the Pr(III) centre, forming a pseudo-ring.

In the title compound, [Ni(C(16)H(8)N(2)O(4))(C(5)H(5)N)(3)]·C(5)H(5)N, the Ni(II) atom is six-coordinated by two O atoms and one N atom from the Schiff base ligand and by three N atoms from three pyridine mol-ecules, forming a distorted octa-hedral geometry. The Ni-O(phenolate) bond [1.9750 (16) Å] is shorter than the Ni-O(carbon-yl) bond [2.0840 (16) Å] and the Ni-N bonds (mean 2.120 Å).

A gas-liquid hybrid pulsed discharge system with a multi-needle-to-plate electrode geometry was used in the present study. A multiphase (gas-liquid-solid) pulsed discharge system was then formed by adding glasses beads immobilized with TiO2 photocatalyst into the discharge system. In the present paper, ultraviolet light produced during the pulsed discharge process was used as the lamp-house to induce the photocatalytic activity of the TiO2 photocatalyst. The synergistic effect of pulsed discharge and TiO2 photocatalysis was reviewed by the spectral diagnosis of hydroxyl radical ( *OH) in the pulsed discharge system. The obtained results showed that the emission spectrum of *OH could be observed at 306 nm (A2Sigma+-->X2II), 309 mn (A2Sigma+ (v' = 0) --> X2II (v" = 0)) and 313 nm (A2Sigma+ (v' = 1) --> X2II (v" = 1) transition). The relative emission intensity of *OH at 313 nm in the discharge system was the strongest among the three characteristic spectra. The relative emission intensity of *OH at 313 nm was stronger by adding TiO2 photocatalyst into the pulsed discharge system than that in the sole pulsed discharge system. In the case of experiments that changing the gas bubbling varieties and initial solution pH values, the results revealed that the relative emission intensity of *OH at 313 nm in the synergistic system was stronger when Ar was used as bubbling gas compared with that when air and oxygen were bubbled into the reaction system. Furthermore, the acidic solution system was favorable for producing more *OH, and therefore the corresponding emission intensity of *OH at 313 nm was stronger than that in the neutral and basic solution.

The severe acute respiratory syndrome-associated coronavirus (SARS-CoV) spike protein (S) is a major target for neutralizing antibody. To develop and apply a safe neutralization assay for SARS-CoV, lentiviral SARS-CoV S pseudotypes had been constructed based on a three plasmid system, which contained pVRC8304 (harboring codon optimized full-length SARS-CoV S protein), pCMV delta 8. 2 (HIV-1 gag/pol construct) and pHR'CMV EGFP (the green fluorescent protein reporter construct). The pseudo-typed lentiviral particles were used to develop an in vitro microneutralization assay that was both sensitive and specific for SARS-CoV neutralizing antibody. We used this assay to determine the titers of the neutralizing antibodies (Nabs) in serum samples from mice immunized with various rVVs expressing different S fragments of SARS-CoV. The serum antibodies derived from S and various segments of S1 region neutralized SARS-CoV in vitro. No cross-neutralization occurred with the goat antiserum prepared with inactivated HCoV-OC43 or HCoV-229E. Neutralization titers measured by this assay were highly parallel with those measured by the assay using live SARS-CoV. Because the pseudotype assay does not require handling live SARS virus, it is a useful tool to determine serum neutralizing titers during natural infection and the preclinical evaluation of candidate vaccines.

Background: The use of camptothecin (CPT) derivatives is limited by severe toxicity associated with the instability of their chemical structure. We have developed a stable CPT derivative (CPT417) and have investigated its biological activity in comparison to the currently used CPT analogs., Materials and Methods: The anticancer effects of CPT417 were assessed in vitro, with glioblastoma and colon cancer cell lines, and in vivo with mice bearing mammary adenocarcinoma tumors implanted subcutaneously in the flank. Cytotoxicity was assessed using vital dye exclusion, timelapse microscopy and colony formation., Results: CPT417 and topotecan inhibited glioblastoma cell growth at comparable levels and both compounds inhibited clonogenicity of colon cancer cells more effectively than irinotecan. CPT417 showed a much greater inhibition of mammary tumor growth compared to topotecan, both by intraperitoneal and oral administration., Conclusion: CPT417 shows dramatically reduced toxicity and an enhanced antitumor activity compared to topotecan.

Objective: To study the therapeutic effect and possible mechanism of total panax notoginseng saponins (PNS) for treatment of rheumatoid arthritis (RA), and to observe its safety and influence on RA immune related inner environment., Methods: Eighty-four patients were randomly assigned to two groups. All were treated with the routine therapy with diclofenac sodium, Leflunomide and prednisone, but for the 43 patients in the treatment group PNS was given additionally. The therapeutic course was 28 days for both groups. Clinical efficacy and change of indexes including platelet counts, immnuoglobulins (IgG, IgA, IgM), complement (C)3, rheumatoid factor (RF), C-reactive protein (CRP), ceruloplasmin (CER), haptoglobin (HPT), and alpha1-acid glycoprotein (AAG) were observed., Results: Significant improvement of clinical symptoms, including the joint swelling index, joint tenderness index, joint pain index, time of morning stiffness and VAS revealed in both groups after treatment, and the effect in the treatment group was better (P<0.05 or P<0.01). PLT, CER, AAG, HPT, CRP, IgG, IgA, IgM, C3 and RF were lowered in both groups (P<0.01), but the lowering in PLT, CER, AAG and CRP in the treatment group was more significant than that in the control group respectively (P < 0.05 or P < 0.01)., Conclusion: PNS can significantly improve the condition of patients, enhance the therapeutic effect in treating RA, through regulating the disordered immunity and improving the effect of anti-inflammatory and analgesia.

In this study the traditional Chinese medicine compound recipe (TCMCR) Shuxiong sustained-release capsules (SXSRC) were prepared by multiparticulate time-controlled explosion technology. First, Shuxiong pellets were prepared with the refined medicinal materials containing in the recipe of Shuxiong tablets. Then, the pellets were coated sequentially with an inner swelling layer containing low-substituted hydroxypropylcellulose as the swelling agent and an outer rupturable layer of ethylcellulose. Finally, SXSRC were developed by encapsulating five kinds of pellets whose respective coating level of outer layer was 0%, 9%, 15%, 18% and 20% at equivalent ratio in hard gelatin capsules. Under the simulated gastrointestinal pH conditions, the in vitro release test of SXSRC was carried out. The value of similarity factor (f2) of hydroxysafflor yellow A and Panax notoginseng saponins, hydroxysafflor yellow A and ferulic acid, Panax notoginseng saponins and ferulic acid was 90.1, 77.3, 87.0, respectively. The release profiles of these three compositions from SXSRC showed a characteristic of obvious sustained-release and no significant difference between them. The results indicated that using multiparticulate time-controlled explosion technology various components in TCMCR with vastly different physicochemical properties could be released synchronously while sustained-releasing. That complies with the organic whole conception of compound compatibility of TCMCR.

The phylogenetic relationship of Shennongjia white bear has been an open question. Total DNA was extracted and sequenced from hair and feces of Shennongjia white bear. Based on the partial Cyt b gene sequence obtained from the samples, the authors aligned them using the Clustal W software program. The MEGA software was used to analyze the divergences and base substitutions of the partial Cyt b gene among the 11 species: Shennongjia white bear, Selenarctos thibetanus, Euarctos americanus, Helarctos malayanus, Ursus arctos, Thalarctos maritimus, Melursus ursinus, Procyon lotor, Ailuropoda melanoleuca, Ailurus fulgens and Tremarctos ornatus. The phylogenetic trees constructed by multiple methods (NJ and MP) supported nearly the same topology. Our molecular results show that the sequence divergence between Shennongjia white bear and Asiatic black bear (Selenarctos thibetanus) is lower than that between other species.

Using our lab's improved protocol for faecal DNA extraction, the entire 753-bp DNA coding sequence of the nuclear brain-derived neurotrophic factor (BDNF) gene was cloned for the first time from Asiatic Black Bear Selenarctos thibetanus faecal samples with primers based on the reported sequence of the Malayan Bear BDNF gene. Hair was used as a positive control and the experiments were repeated several times to obtain reliable and identical results. Sequence analysis showed that the BDNF gene of Asiatic Black Bear was highly conserved compared to those of human and giant panda, with an identity of 94.5% and 98.9%, respectively. The deduced amino acid sequence of the mature protein was found to be identical to those of all the reported mammalians. According to gene sequence alignment, the giant panda appeared to be phylogenetically closer to Asiatic Black Bear than the lesser panda. This study represents the first time that a non-invasive method such as faecal sampling was used to analyze a functional nuclear BDNF gene of Asiatic Black Bear. It will not only provide important reference for the conservation and breeding of Asiatic Black Bear and open up new avenues of non-invasive sampling in the study of endangered wildlife, but also provide another molecular evidence for the study of relationship of Asiatic Black Bear and its related species.

Aim: To evaluate in vitro release of lycopene microcapsules. Pharmacokinetic parameters of lycopene microcapsule and lycopene powder as reference were estimated after a single dose of oral administration to dogs. The relationship between in vitro dissolution and in vivo absorption was investigated., Methods: The content of lycopene in the release medium was determined by UV spectroscopy method. Health hybrid male dogs were used as experiment subjects and lycopene powder used as standard to estimate the pharmacokinetics of lycopene microcapsules. HPLC method was used to assay the concentration of lycopene in dog plasma. Pharmacokinetics parameters were estimated by 3P87 program. The drug release percentage in stimulated intestinal fluid was compared with the absorption at a given time point., Results: The release profiles of lycopene from microcapsule showed that the lycopene gelatin microcapsule exhibited enteric property. The pharmacokinetics parameters estimated after oral administration of lycopene powder and lycopene microcapsule in a single dose of 2.5 mg x kg(-1) body weight to dogs were 7.30 h, 15.06 h for T1/2alpha; 28.10 h, 46.76 h for T1/2beta; 22.32 h, 41.03 h for T(max); 1.67 microg x h x L(-1), 2.08 microg x h x L(-1) for AUC(0-infinity), respectively. The concentration-time curves could be fitted to a two-compartment model for both the lycopene powder and the lycopene microcapsule analyzed by 3P87 program. The relationship between in vitro dissolution and in vivo absorption was found to have good correlation (r = 0. 981 9) was found., Conclusion: It could be concluded that lycopene microcapsule was a sustained release dosage form. The result of release in vitro could be used to predict the absorption in vivo.

Taxol and taxotere are two of the most promising anticancer drugs. To determine the mechanisms responsible for cell death after exposure to low doses of taxane, PC3 cells were treated with taxol and taxotere, and observed with immunofluoroscence microscopy. Pericentriolar material dissociation and blockage of normal centrosome separation were found to result in two different abnormal spindle types; multipolar and monopolar spindles, respectively. The majority of abnormal spindles induced by taxol were monopolar spindles, whereas taxotere mostly induced abnormal multipolar spindles. Consequently, monopolar spindle mitosis resulted in cleavage failure, while multipolar spindle mitosis led to the formation of both cleavage failure and multipolar cell division. Multinucleation characterized interphase cells which had undergone cytokinesis defects. These cells subsequently became giant multinucleated cells after several rounds of cell cycle with sustained cleavage failure, and were gradually eliminated through cell death.

Aim: To evaluate the effects of surfactants on the pharmacokinetics and distribution in rats after intravenous administration of SOD liposomes., Methods: The liposomes were prepared by reverse phase evaporation method. The activity of SOD was assayed by method of xanthine oxidase., Results: The T1/2 of SOD solution, common SOD liposome, SOD liposomes modified by DSPE-PEG2000 and Tween 80 were 0.25, 0.34, 0.66 and 0.41 h, respectively; AUC were 12.48, 24.66, 41.16 and 33.02 microg x h x mL(-1), respectively. Compared with the common liposome, the liposomes modified by DSPE-PEG and Tween 80 decreased the content of SOD in liver and spleen, but increased in brain., Conclusion: The three kinds of liposomes could increase T1/2 and AUC in some extent, especially in PEG-L group. Tween-L could increase the SOD content in brain, and PEG-L could decrease the SOD content in the liver and spleen compared with the common liposome.

Aim: To Study the interaction of macrophage colony-stimulating factor (M-CSF) and interleukin-10 (IL-10) in productions of IL-12 and IL-18 and expressions of CD14, CD23, and CD64 by human monocytes., Methods: Purified adherent human monocytes were cultured with M-CSF or IL-10 alone, or with M-CSF+IL-10 and 2-3d later, the culture supernatants and cells were separated and collected. IL-12P40 and IL-18 levels in the supernatants were determined by ELISA and the percentages of CD14, CD23, and CD64 positive cells were examined by flow cytometry., Results: (1) IL-10 decreased M-CSF-induced IL-18 levels, while M-CSF further reduced IL-12P40 level in the culture supernatants of IL-10-treated monocytes; (2) IL-10 alone had no effect on the percentage of CD14-positive cells, but further increased the percentage of CD14-positive cells induced by M-CSF; M-CSF alone had no effect on the percentage of CD64-positive cells, but further increased the percentage of CD64-positive cells induced by IL-10; (3) IL-10 decreased the percentage of CD23-positive cells induced by M-CSF., Conclusion: Between M-CSF and IL-10, there were antagonistic effects on inducing IL-18 and CD23 expressions by monocytes; there were also synergistic effects on inhibiting IL-12P40 production and inducing CD14 and CD64 expressions by monocytes., (Copyright 2004 Acta Pharmacologica Sinica)

Objective: To determine the role of Rho associated kinase (ROK) in contraction of rat femoral arteries after 14 d tail suspension., Method: Male Wistar rats were randomly divided into control group (CON) and 14 d -30 degrees tail suspension group (TS14d). Analysis was performed on the contractile responses of perfused femoral arterial rings from both TS14d and CON rats to KCl, phenylephrine (PE), and U-46619 in the presence of Y-27632., Result: Arterial rings from TS14d rats displayed a reduced contractile response to KCl and PE but not significantly to U-46619. In the response to KCl, Y-27632 caused a concentration-dependent relaxation and significantly larger decrease of contraction in tissues from TS14d rats. Y27632 nearly abolished the tonic component of KCl-induced contraction when its dose was increased from 10(-6)mol/L to 10(-5) mol/L. It had also an inhibitive effect on the PE and U-46619-induced contraction and caused significantly larger decrease in U-46619 but not in KCl or PE induced contraction in tissues from TS14d rats., Conclusion: ROK activity may be enhanced and play a compensational role in rat femoral arteries after TS14d.

Aim: To survey the morphology and size distribution of silymarin microemulsion, to investigate the absorption of silymarin microemulsion in rat intestine compared with the absorption of silymarin micelle in rat Jejunum., Methods: The intestine in rats was canulated for in situ recirculation., Results: The absorption rate constants (Ka) of silymarin microemulsion at the entire intestine, ileum jejunum, duodenum and colon were 6.22 x 10(-2), 2.27 x 10(-2), 1.9 x 10(-2), 1.9 x 10(-2), 1.05 x 10(-2) and 0.43 x 10(-2) h(-1), respectively. The absorption rate constants of two kinds of silymarin micelle at jejunum were 0.36 x 10(-2), 0.65 x 10(-2) h(-1)., Conclusion: Silymarin microemulsion was well absorbed at the middle and lower segments of intestine in rats. The absorption was a first-order process with the passive diffusion mechanism.

This study aimed to investigate the apoptotic effects of novel paclitaxel analogs on NCI/ADR-RES breast cancer cells. Using the colony formation assay, the cytotoxicity of three novel paclitaxel analogs were evaluated on NCI/ADR-RES cells which overexpress multidrug-resistant gene (MDR1). All three novel paclitaxel analogs exhibited significantly higher cytotoxicity on NCI/ADR-RES cells than paclitaxel. One analog, TL139, was 140 times more effective than paclitaxel. Using TUNEL and DNA fragmentation assay, remarkably increased apoptosis in the paclitaxel analog-treated cells was observed at 48-72 hours, but not in paclitaxel-treated cells. Caspases-3/7 were dramatically activated at 48-72 hours by the novel paclitaxel analogs. The enhanced activity of caspases-3/7 was evidently verified by the measurement of the cleavage of poly(ADP-ribose) polymerase (PARP). The increased activity of caspases-3/7 significantly correlated with the enhanced apoptosis and cell survival data. Treatment with paclitaxel analogs resulted in a significant amount of mitotic arrest. Using Western blot, the phosphorylation of Bcl-2 protein was found in palictaxel analog-treated cells in a time-dependent manner similar to that of mitotic arrest, thereby indicating that there existed a close correlation between Bcl-2 phosphorylation and mitotic arrest that preceded apoptosis. We conclude that novel taxane analogs could effectively kill MDR1-positive breast cancer cells via the mode of apoptosis.

Objective: To investigate the effect of thalidomide on bone marrow cells gene expression in multiple myeloma (MM) patients with suppression subtractive hybridization (SSH) and explore the molecular mechanism of thalidomide therapy for MM., Methods: In a MM patient receiving thalidomide therapy and bone marrow cell from himself, total RNA extraction, mRNA isolation and cDNA synthesis were carried out respectively with routine procedures. SSH were performed in A and B group respectively. The subtracted cDNA was selectively amplified by suppression PCR. The product was inserted into T vector, and then transfected into the competent host JM109. So two subtractive libraries were constructed. After blue-white screening, colonies were selected and plasmids extracted. Homologous comparation was conducted in GenBank., Results: In group A, seven clones were isolated, including ribosomal protein L19 (HUMAN), IgG lambda chain v-v region (HUMAN), contains Alu repetitive element, NADH-ubiquinone oxidoreductase chain 2, elongation factor 1-gamma, human beta globin region, and 40S ribosomal protein S4 (HUMAN). In group B, six clones were isolated, including cytochrome B, up-regulated by 1,25-dihydroxyvitamin D(3) (VDUP1), NADH- ubiquinone oxidoreductase 20 Kd subunit, mu-calpain large subunit, tumor protein, translationally-controlled 1 (TPT1) and COATOMER alpha subunit., Conclusion: Thalidomide induces apoptosis and antiangiogenesis by down-regulating some genes and up-regulating some others genes.