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To explore the relation between apolipoprotein CIII (ApoCIII) gene polymorphism and coronary artery disease (CAD).Peripheral blood samples were collected from 400 healthy blood donors and 360 CAD patients. Total cholesterol (TC), triglycerides (TG), high-density lipoproteins cholesterol (HDL-C), low-density lipoproteins cholesterol (LDL-C), and ApoCIII were detected. Fluorescence quantitative PCR was used to detect the ApoCIII) gene polymorphisms. Genomic DNA was extracted from whole blood samples. We study the relation between variants of ApoCIII-625 and -455 and CAD by FQ-PCR. Then we explore the relation between variants of ApoCIII-625 and -455 and the concentration of lipid. All objects were measured about Total Cholesterol (TC), Triglycerides (TG), high-density lipoproteins cholesterol (HDL-C), low-density lipoproteins cholesterol (LDL-C) and ApoCIII.The ApoCIII-455C homozygote rate of the CAD patients was 70.1%, significantly higher than that of the healthy persons (53.1%). The ApoCIII-625del-455c phenotype rate of the CAD patients was 49.3%, significantly higher than that of the healthy persons. The ApoCIII-455C expression rates of the patients with hyperlipidemia and those with hyperlipidemia combined with CAD were high. There was no significant difference in the expression of ApoCIII-455c between the CAD patients and the normal persons.The gene polymorphisms of ApoCIII-455 increase the risk of CAD. ApoCIII-455C gene is positively related to the risk of CAD. The homozygotes of the ApoCIII-455C gene have high risk of CAD. ApoCIII-455C in cooperation with -625 (del) will increase the risk of CAD.

Chicken embryo fibroblast (CEF) is a primary cellular material to research the infectious bursal disease virus (IBDV). Constructing the cDNA expression library of CEF is the foundation to research cell tropism and find cell receptors of IBDV from CEF. In order to achieve that purpose, a high-quality cDNA expression library of CEF was constructed by Gateway technology, which could avoid using the restriction enzyme for cloning to solve technical limitation of roution method. The mRNA was extracted from chicken embryonic fibroblast. Moreover, single-strand cDNA and double-strand cDNA were synthesized by using biotin-conjugated Oligo (dT) primer in turn. The double-strand cDNA was ligated Adapter and then purified by the cDNA Size Fractionation Columns. After BP recombination reaction, a cDNA entry library was constructed with a titer of 1 x 10(6) cfu/mL, total clones of 1.2 x 10(7) cfu and an average insertion size of about 2243 bp. After LR recombination reaction, the cDNA entry library was transformed into expression library which took on a titer of 5 x 10(5) cfu/mL, total clones of 5.5 x 10(6) cfu and an average insertion size of about 2411bp. The results indicate that the constructed cDNA expression library performs a remarkable high value in both recombination rate and library coverage. As a result, the cDNA expression library, with its good quality, may facilitate to identify the receptors associated with the resistance against IBDV in chicken embryonic fibroblast and to cast new light on the mechanism of cellular tropism. Moreover, it may also provide data of chicken embryonic fibroblast in transcription level and may be helpful to study its biological functions.

The gamma-linolenic acid-producing fungus Mortierella isabellina 6-22 is an important industrial strain. To clarify the biosynthetic pathways for polyunsaturated fatty acids in this strain, a disruption vector pD4MI6, including 5' and 3' regions of the fatty acid Delta(6)-desaturase open reading frame as homologous recombination elements and the Escherichia coli hygromycin B (HmB) phosphotransferase gene (hph) as selectable marker, was successfully constructed. Following transformation of pD4MI6 into the hygromycin B-sensitive recipient strain M. isabellina 6-22-4, a Delta(6)-desaturase gene-defective mutant strain was selected that was unable to produce gamma-linolenic acid as determined by gas chromatography and molecular analysis. The morphology and physiology of the mutant, such as colony shape, color, and growth rate, were changed dramatically compared with that of strain M. isabellina 6-22-4.

To study the effect of Jinguo Weikang Capsule [see text] on the gene expression of H-ras, epidermal growth factor receptor (EGFR), P53 and C-myc of the gastric mucosa in rats with gastric precancerous lesions, and to investigate the action mechanism of JWC on gastric precancerous lesions.A rat model with paratypical proliferation of the gastric epithelium mucosa was established by using 60Co irradiation. Rats were divided into the normal group, model group, high-, medium-, low-dose JWC treatment groups, and the vitacoenzyme control group, and were treated for 30 days. The expression of H-ras, EGFR, P53 and C-myc genes of the gastric mucosa was detected by using immunohistochemical methods.The expression and over-expression rates of H-ras, EGFR, P53 and C-myc gene in the high-and medium-dose JWC treatment groups were significantly lower (P0.05) as compared with those of the model group.JWC can inhibit the expression of the H-ras, EGFR, P53 and C-myc genes expression of the gastric mucosa in rats, which may be one of mechanisms involved in suppressing or reversing gastric carcinogenesis.

A strain Mortierella isabellina M6-22-4, which was sensitive to hygromycin B, was selected by treating parental spores with N-methyl-N' -Nitro-N-nitrosoguanidine (MNNG). Protoplasts of the strain Mortierella isabellina M6-22-4 were transformed successfully to hygromycin B resistance using the PD4 plasmid, which contains the Escherichia coli hph gene under the control of Mortierella alpina his H4.1 promoter. The PD4 plasmid was introduced by PEG/CaCl2 treatment. Transformation frequencies of 1.6 - 2.8 transformants/microg of DNA were achieved. Then they were successively incubated to non-selected PDA plates for 10 generations. About 31.6% transformants only from digested plasmid were mitotically stable and showed different hygromycin B resistance when they were incubated back to selection plates. The results of PCR and Southern analysis in three transformants indicated that the plasmid PD4 had been integrated into the fungal genome with 1 - 2 copies. This is the first report of Mortierella isabellina transformation system and supplies an important tool for further research into genetic manipulation of this filamentous fungus.

The coding region of VP2 gene from Chicken Infectious Anemia was amplified from genome extracted from chicken liver tissue by PCR. PCR product was double digested with restriction enzymes BamH I and Sal I and cloned into pET-28a digested with BamH I and Sal I. Subsequently, the recombinant plasmid pET-28-VP2 was extracted and double digested with restriction enzymes BamH I and Sal I. After confirming its rightness by PCR and analysis of restriction endonucleases, the recombinant plasmid pET-28-VP2 was transformed into E. coli BL21 (DE3) strain. The culture was induced by 1 mmol/L IPTG at 37 degrees C for three hours and analyzed with SDS-PAGE. The result shows that gene encoding VP2 of CIAV was expressed successfully in E. coli and the fusion protein existed in supernatant, which was about 31kDa and showed specific immunoreactivity with anti-CIAV sera in Western blot. The fusion protein was purified by Ni2+ -affinity chromatography and quantitated by Bradford method. Then BALb/c mice were immunized with purified protein emulsified with Freund's complete adjuvant on day 0 and boosted twice on day 14 and 28 with the same dose of antigens emulsified with Freund's incomplete adjuvant, respectively. The serum isolated were examined by an enzyme-linked immunosorbant assay (ELISA) using the purified VP2 and CIAV as coating antigens and the serum could react with target protein and CIAV in ELISA detection test.

Following the requirements of the "National survey on the current status of the major human parasitic diseases", the investigation was conducted in June-October 2002 in 5 districts (counties) of Beijing with a sample of 7912 people. The overall prevalence rate of intestinal parasites was 2.9%, significantly lower than the result from the first survey in 1988-1989 (34.8%) (chi2=3 227.45, P0.05), revealing that intestinal parasitic infections are not an important risk for people in Beijing Municipality in general.

Tunable diode laser absorption spectroscopy has been applied in the fields of atmospheric chemistry and monitoring pollutant gases as a new method of measuring trace gases. The technique of remote sensing of CO and CO2 at 760 mm Hg pressure with tunable diode laser absorption spectroscopy in the near-infrared region is introduced. And the relationship between the second-harmonic spectrum of CO2 in Lorentzian line shape and the modulation amplitude is also presented.

To evaluate the effectiveness of directly observed treatment (DOT) implementation in tuberculosis (TB) control in China.Systematic literature review was carried out for published and unpublished articles regarding DOT, and meta analysis was used to compile results from selected papers. Also, sensitive analysis was carried out to explore the potential factors influencing the effectiveness of DOT.Two hundred and four articles were identified from China National Knowledge Infrastructure, Wanfang and Medline databases during 1994 - 2004 in which 120 articles were eligible according to our selection criteria. (1) Regarding TB case management: TB patients managed under DOT was compared to cases managed under the whole course treatment management (WCM). The combined RD of cure rate was 0.14 (95% CI: 0.06 - 0.22) and the two case management methods were significantly different (P0.00001). Data from stratified analysis showed that strict implementation of WMC had similar effectiveness with DOT (P = 0.06), and with RD 0.02 (95% CI: 0.00 - 0.03). However, the cure rate of DOT management was significantly better than that under un-strict WCM management (P0.05) with RD 0.16 (95% CI: 0.07 - 0.24). The cure rate of DOT was significantly better than those patients under self-administration (RD was 0.24 with 95% CI: 0.10 - 0.38 and P = 0.002). (2) Chemotherapeutics: when short-course and long course-treatments were compared, the combined RD became 0.04 (95% CI: 0.01 - 0.07) and P = 0.003.If the implementation was under strict management, the different models of patient management showed similar adherence rates. However, the effect of adherence would depend on the regimen, pattern of intake of the drugs and the methods of supervision being used.

One-photon fluorescence(OPF) and two-photon fluorescence (TPF) of two chromophores 1,4-bis(9-ethyl-vinyl carbazole) naphthalene (abbreviated as BECVN) and 1,4-bis(4-N, N-methylamino styryl) naphthalene (abbreviated as BMABN) were resported. Strong emission at 492-541 nm and at 500-556 nm, respectively, were obtained under the excitation of -370 nm (Xe lamp) and of 730-800 nm femto-second laser pulses, respectively. Although the relative fluorescence quantum yield (phi(f)) of BECVN is as high as 7.4 times that of BMABN, the latter possesses higher TPF intensity and larger two-photon absorption (TPA) cross-section, suggesting that BMABN containing naphthalene as pi-center end-capped with N,N-methylamino styryl group exhibits more effective two-photon process. Lippert equation showed that the difference of dipole momnet (deltamu(ge)) between the ground state and the excited state for BECVN is nearly twice that for BMABN, conforming to that the deltamu(ge) value is not the key for the enhancement of two-photon absorption cross-section.

To compare the prognostic value of different electroencephalography (EEG) grading standards of cerebral dysfunction after cerebrovascular diseases.The EEG of all the 40 patients who suffered from acute cerebrovascular disease of the cerebral hemisphere were recorded continuously, and three different EEG grading standards (Lavizzari, Synek and Young) were analyzed and compared.All three different EEG grading standards showed significant correlation with outcome (all P0.001), i. e. the higher the EEG grade was, the poorer the outcome was. Logistic regression showed that the Synek standard was the best for survival rate and comprehensive prognostic accuracy (90.9%, 89.2%, respectively), Young standard (81.8%, 83.8%) was lower than Synek standard in predicting accuracy but higher than Lavizzari standard (75.%, 80.% respectively).The EEG grading standard of Synek might well reflect the cerebral dysfunction and appear to be a reliable predictor for outcome.

Incubate golden hamster spermatozoa and NF (kappa) B-luc(+) in co-culture and liposome mediated methods, and then process the IVF with oocytes, And two groups of samples were detected by PCR, Southern Blot and FISH. Analyzed the relative data by statistics. The results showed that spermatozoa of Golden hamster epididymis can take the foreign DNA voluntarily, and the foreign DNA exist in the head of the spermatozoa can enter into zygotes by IVF. The exist of liposome can improve the foreign gene transfect ratio and transgenic ratio.

We present a modified \emph{susceptible-infected-susceptible} (SIS) model on complex networks, small-world and scale-free, to study epidemic spreading with the effect of time delay which is introduced to the infected phase. Considering the topology of the network, both uniform and degree-dependent delays are studied during the contagion process. We find that the existence of time delay will enhance both outbreaks and prevalence of infectious diseases in the network., Comment: 10 pages, 5 figures. to appear in Physica A

To evaluate the effect of dynein inhibitor on mouse oocyte in vitro maturation and its cyclin B1 transcription level.Immature mouse oocytes were cultured in vitro with a known dynein ATPase activity inhibitor-sodium orthovanadate (SOV) to detect the changes of maturation rate, and semi-quantitative RT-PCR and single cell RT-PCR were performed to detect the changes of cyclin B1 mRNA level.In dose-dependent experiments, the maturation rates of oocytes were significantly different between 5 micromol/L SOV and control groups (P0.05), and decreased with SOV increasing doses. However, the elevation of cyclin B1 mRNA level of immatured oocytes cultured for 12 h depended on SOV concentrations ranging from 50 to 500 micromol/L. In incontinuity exposed SOV experiments, the maturation rates of oocytes markedly reduced after the first incubation with 400 micromol/L SOV at least for 1 h and were first cultured in SOV-free medium for 4 h or 8 h before exposure to SOV (P0.05). In time-course experiment, the opposite changes of cyclin B1 mRNA level in oocytes between SOV and control groups were observed.Dynein inhibitor might delay oocytes meiosis process, and cause ectopic expression of cyclin B1 in oocytes. Most Oocytes incubated with SOV blocked at germinal vesicles (GV) stage or M I to anaphase transition due to dynein dysfunction and ectopic transcription level of cyclin B1.

To evaluate the reporting and transferring system for tuberculosis (TB) patients carried out by city General Hospitalstownship clinics in the last 10 years in China.Systematic review and Meta-analysis were carried out.After 5 years of follow-up on the outcomes of intervention, it was found that both the rates of newly registered smear positivity and the new case registration on smear positivity had significantly been increasing during the last five years and the scale of increase was growing annually during the first three years. However, the scale of increase started to decline on the fourth year. The combined RR on the failure of transfer rate was 0.36 (95% CI: 0.25 - 0.53).The integrated outcomes showed that the ongoing reporting and transferring system in general hospital was benefit for TB case detection.

The thermophilic microorganisms (50-60 degrees C) are introduced and the contact oxidation technology is adopted to treat with the high temperature effluents from an oil field in this paper, which solves a problem that the effluents need to be cooled in routine biotreatment. The results of the pilot test show that the HRT of the biotreatment system adjusted to 8-18h can make CODCr of the effluents stable at 175-263mg/L and oil content under 10mg/L on condition that the temperature of the influents keeps at 60-65 degrees C, CODCr 300-590mg/L and the quality of the oil wastewater is greatly fluctuant. Flocculating process, then, can effectively flocculate and precipitate the suspending and residual chroma pollutant from the effluents of biotreatment and CODCr in the effluents of flocculating process can reduce to 136-229mg/L. And CODCr even reduces to under 150mg/L after flowing by sand and activated carbon filtration equipments. Under the high temperature condition, furthermore, combined stuffing is an ideal carrier for the thermophilic microorganisms.

To investigate the methods of evaluating brain death accurately and objectively.Various kinds of clinical examination [including conscious state, Glasgow coma score (GCS), brain stem reflection, cranial nerve dominated actions, spinal reflection, spinal nerve dominated automatic action, respiratory and apnea testing] and laboratory methods [such as electroencephalography (EEG), brainstem auditory evoked potentral (BAEP), brainstem auditory sensery evoked potentral (SLSEP) and transeranial Doppler (TCD)] were used to evaluate 11 cases with brain death.Eleven cases presenting with deep coma were conssedered to have brain death. All of them had GCS score of 3 and disappearance of brain stem reflection and cranial nerve dominated automatic action. Six of them patients showed anisocoria. Nine of them pharyngeal reflex could not be detected because of tracheal cannulization, but this did not influence the evaluation of brain death. None of them showed "flat" EEG and disappearience of BAEP dominant wave. None of the SLSEP show any wave behind N13. TCD showed blood stream characteristic of brain death except in one case blood stream characteristic of increased intracraniai pressure. Apnea test and atropine test provided the final proof brain death. The former had more influence on heart rate, blood pressure and oxygen saturation while the latter were more safe and reliable.Combined and continuous observation of clinical and laboratory indices can enhance the accuracy of the evaluation of brain death.

The neuroprotective effect of docosahexaenoic acid (DHA) on the glutamate-induced cytotoxicity in rat hippocampal cultures was investigated in the present study. DHA at 5-50 microg/ml successfully protected neurons against the cytotoxicity, markedly increased the cell viability, inhibited both nitric oxide (NO) production and calcium influx, and increased the activities of antioxidant enzymes of glutathione peroxidase (GSH-Px) and glutathione reductase (GR). However, it did not alter the levels of glutathione (GSH) as compared to the control. These results suggest that DHA might be a potent neuroprotector. In addition, they may help to improve our understanding of the effect of DHA on neurodegeneration.

To investigate the effect of ionizing radiation on the expression of p16, CyclinD1, and CDK4 in mouse thymocytes and splenocytes.Fluorescent staining and flow cytometry analysis were employed for the measurement of protein expression.In time course experiments, it was found that the expression of p16 protein was significantly increased at 8, 24, and 48 h for thymocytes (P0.05, P0.01, and P0.05, respectively) and at 24 h for splenocytes (P0.05) after whole body irradiation (WBI) with 2.0 Gy X-rays. However, the expression of CDK4 protein was significantly decreased from 8 h to 24 h for thymocytes (P0.05-P0.01) and from 8 h to 72 h for splenocytes (P0.05-P0.01). In dose effect experiments, it was found that the expression of p16 protein in thymocytes and splenocytes was significantly increased at 24 h after WBI with 1.0, 2.0, and 4.0 Gy (P0.05-P0.01), whereas the expression of CDK4 protein was significantly decreased with 2.0 Gy for thymocytes (P0.05) and 0.5-6.0 Gy for splenocytes (P0.05-P0.01). Results also showed that the expression of CyclinD1 protein decreased markedly in both thymocytes and splenocytes after exposure.The results indicate that the expression of p16 protein in thymocytes and splenocytes can be induced by ionizing radiation, and the p16-CyclinD1/CDK4 pathway may play an important role for G1 arrest of thymocytes induced by X-rays.

Photochemotherapy employing 8-methoxypsoralen and long-wavelength ultraviolet radiation (UVA, 320-400 nm) is widely used in the treatment of psoriasis. The photoactivation of psoralens in skin cells leads to formation of DNA photoadducts which may be responsible, at least in part, for the efficacy of these photochemotherapies. However, mutations arising from these adducts may also lead to the well-characterized increased incidence of squamous cell carcinoma. Mutations in the p53 tumor suppressor gene have been detected in many human cancers. To determine whether p53 mutations occur in squamous cell carcinomas in PUVA patients, PCR was used to amplify the exons (5-9) in which other studies have found a high frequency of point mutations. Gel electrophoresis was used to detect single-strand conformational polymorphisms. Aberrantly migrating bands were excised, reamplified and sequenced. Thirty-four specimens from 10 patients were examined. Specimens from one patient who had received no phototherapy as well as from normal controls were also analyzed. Five of the 10 patients showed at least one p53 mutation. In contrast to previously reported psoralen-induced p53 mutations in mice, the expected psoralen type mutations at alternating AT sites were not detected. All but two of the altered sequences occurred at dipyrimidine sites which is typical of solar type mutations. Two C-->T mutations and two dipyrimidine mutations (CC-->TT) were found. Other mutations included: C-->G, G-->T, C-->A and an 18 bp deletion. A review of therapeutic history of these patients showed that some had also received UVB phototherapy. Furthermore, because sunlight is thought to be beneficial for psoriasis, nontherapeutic, casual UVB exposure cannot be excluded. Our observations suggest that the SCC may have arisen from the solar mutations and that PUVA may enhance tumor progression or immune suppression.

To investigate the risk factors of scrub typhus infection in Beijing, China, a case-control study was carried out. Cases (n = 56) were defined as persons who were diagnosed by PCR and serological method within three years. Three neighborhood control subjects were selected by matching for age and occupation. Living at the edge of the village, living in the houses near grassland, vegetable field or ditch, house yard without cement floor, piling weeds in the house or yard, all of these were risk factors for scrub typhus infection. Working in vegetable fields and hilly areas, and harvesting in autumn posed the highest risks, with odds ratios (ORs) and 95% confidence intervals (CIs) of 3.7 (1.1–11.9), 8.2 (1.4–49.5), and 17.2 (5.1–57.9), respectively. These results would be useful for the establishment of a detail control strategy for scrub typhus infection in Beijing, China.

This study presents a novel method that direct intramyocardial injection of low-dose plasmid DNA and microbubbles combined with insonation could further augment gene expression in normal and ischemic canine myocardium. Plasmids encoding enhanced green fluorescent protein (pEGFP) and hepatocyte growth factor (pHGF) (500 μg) were individually mixed with 0.5 ml of microbubble solution (MB) and injected into the normal or acute ischemic canine myocardium. The dogs in the plasmid + MB/US group underwent insonation (US). Other dogs were randomly divided into three treatment groups: plasmid and insonation, plasmid and MB injection, and plasmid injection only. The EGFP and HGF mRNA expressions were assessed in the myocardium at the injection site and at sites 0.5 and 1 cm remote from the injection site. Compared to plasmid transfer alone, a mean 13.4-fold enhancement of gene expression was achieved in the EGFP + MB/US group at 48 h (p

The title compound, C(20)H(32)ClNO(3), was obtained via a tandem asymmetric Michael addition-elimination reaction of (5S)-3,4-dichloro-5-(l-menth-yloxy)furan-2(5H)-one and 4-methyl-piperidine in the presence of potassium fluoride. The furan-one ring is approximately planar [maximum atomic deviation = 0.022 (2) Å] while the cyclo-hexane ring adopts a chair conformation. Weak inter-molecular C-H⋯O hydrogen bonding is present in the crystal structure.

To observe the effect of Xuesaitong (XST) soft capsule in treating postpartum hemorrhage. Sixty parturients were randomly divided into the treated group (A) and the control group (B), 30 cases in each, and they were treated with XST and placebo respectively from the delivery day for 7 days to observe the changes of hemorrhagic conditions. After delivery, the hemoglobin, RBC count, hematocrit and cardiac index were lower at 3rd day than those before delivery in both groupsP