OBJECTIVE Toestablisharecombinantcelllinethatcanstablyexpressratpurinergic P2receptorP2X4(rP2X4R).METHODS ToconstructgreenfluorescentproteinandrP2X4recombinant plasmid (pEGFP-N1-rP2X4),lipofectamine was used to transfect pEGFP-N1-rP2X4 into human embry-onic kidney (HEK293)cells that were screened with G41 8 (1 g·L-1 ).The quantitative expression of rP2X4 receptor was verified by qRT-PCR and Western blotting analysis.Whole-cell patch clamp record-ing was used to investigate the function of the stably expressed rP2X4 receptor. RESULTS The sequence of plasmid pEGFP-N1-rP2X4 was verified by PubMed Blastn comparison.qRT-PCR and Western blotting analysis demonstrated that the expression of rP2X4 receptor in HEK293-pEGFP-N1-rP2X4 cell lines remained stable after 25 generations (P1 ,P3,P5,P10,P15,P20 and P25).Whole-cell patch clamp recording experiments showed that the rP2X4 receptor agonist,purine-5′-triphosphate (ATP,3.0 μmol·L-1 ),could activate rP2X4 receptors in HEK293-pEGFP-N1-rP2X4 cell lines.Specific activating current could be blocked by non-selective rP2X4 receptor antagonist TNP-ATP (30.0μmol·L-1).CONCLUSION rP2X4receptorisstablyexpressedinHEK293-pEGFP-N1-rP2X4cell line and maintains stable expression and function within 25 continuous generations.The establish ment of HEK293-pEGFP-N1-rP2X4 cell line can contribute to further investigations of the roles of rP2X4 receptors in neuropathic pain.

ObjectiveTo investigate the recent development of Acinetobacter baumannii infection and its drug-resistance.MethodsThe Acinetobacter baumannii infections of inpatients during the year 2001 to June,2005 were analyzed retrospectively. ResultsThe infection of Acinetobacter baumannii increased from 15 strains in 2001 to 68 strains in the first half of 2005.The rate of high drug-resistance strains also raised from 20% to 77%.ConclusionThe incident of infection of Acinetobacter baumannii continuously increased in the past 5 years and became more difficult to treat.

ObjectiveTo investigate the clinical features of chronic lung diseases patients combined with lung fungal infection.MethodsThe data of 216 hospitalized cases with chronic lung diseases were retrospectively analyzed.ResultsThe rate of lung fungal infection of chronic lung diseases patients was 28.1%, and the main pathologic fungal was Candida albicans, about 67.2 %.ConclusionThe chronic lung diseases patient has a higher rate of lung fungal infections compared with other diseases. The measures of preventing, diagnosing and treating lung fungal infection at early stage should be taken.

Objective To investigate the effects of preoperative parecoxib on the levels of inter-leukin 6 (IL-6)and postoperative analgesia for breast surgery.Methods Sixty breast cancer patients undergoing mastectomy were randomly divided into two groups:parecoxib group (group P)and con-trol group (group C),n=30 in each group.All patients received sevoflurane and fentanyl anesthesia. Group P was injected parecoxib 40 mg at 10 minutes before induction of anesthesia,meanwhile group C was injected saline 5 ml.All patients received postoperative patient-controlled intravenous analgesia (PCIA)with fentanyl.VAS scores for pain were assessed at postoperative 2,4,8,12,24 hours. The serum levels of IL-6 were measured by ELISA at 10 minutes before induction,4 h,8 h,and 24 h after surgery.Results Group P had lower VAS scores than group C at 2-12 h after surgery (P <0.05).Compared with 10 minutes before induction,the levels of IL-6 increased significantly at post-operative 4,8,24 h in two groups (P<0.01),while group P had lower levels of IL-6 than group C (P <0.01).Conclusion Preoperative administration of parecoxib has a stronger analgesic effect in breast cancer patients after mastectomy,and decreases the levels of IL-6.

ObjectiveTo investigate the diagnosis and correlative factors of suspected severe acute respiratory synd rome (SARS) cases, and finally, complete the diagnosis and treatment.MethodsTo analyze the clinical, laboratory data,chest radiog raph and treatment of 100 suspected SARS cases.ResultsAmong 100 suspected SARS cases,66 were diagnosed as SARS,4 were mycoplasma l pneumonia, 2 were pulmonary tuberculosis,the others were general bacteria pne umonia.Conclusions SARS hasn't characteristic clin ical manifestation.Because there is not a gold diagnosis standard up to now, the disinfection and isolation of suspected SARS cases should be treated as SARS c ases. In order to prevent missed and misdiagnosis, the diagnosis and treatment s hould be synthetically thought.

Objective To investigate the mechanism and significance of low concentration nitric oxide (NO) inhalation in the treatment of pulmonary thromboembelism in swine. Method The pulmonary thromboem-bolism(PTE) model was made in 15 healthy infantile swines which were subsequently assigned to either control group (n = 8) or NO group (n = 7). Swines of the control group were not treated with any medicine, while 10 ppm of NO was administered by continuous inhalation for 2 hours to swines of NO group. Volume of physiological dead space (VDphy), volume of alveolar dead space (VDalv), intrapulmonary shunt (Qs/Qt), mean pulmonary arterial pressure (PAP), systolic blood pressure (SBP), heart rate (HR), cardiac output (CO), arterial blood pH (pH), arterial partial pressure of carbon dioxide (PaCO2) and arterial partial pressure of oxygen (PaO2) were measured 30 min before and 0 min, 30 min, 60 min, 120 min and 180 min after establishment of VIE. Results The post-FIE VDphy, VDalv, Qs/Qt and PAP in both groups increased markedly after PTE compared with the cor-responding pre-PTE measurements (P < 0.01). Post-FIE PaO2 of both groups decreased significandy (P <0.05 and P <0.01), while significance difference was found between pre- and post-PTE HR, SBP, CO, pH or PaCO2 in neither groups (P > 0.05). Both post-PTE PAP and VDalv in NO group were markedly lower(P <0.05 and P <0.01) and beth PaCO2 and PaO2 were much higher than those of the control group (P <0.05). No signi-fieant difference were found in other measurements between two groups. Conclusions Pulmonary arterial pressure may be lowered, alveoli dead space may be reduced and PaCO2 increased by low concentration NO inhalation for the treatment of PIE without decline in haemodynamic status.

OBJECTIVE To establish a new cell line that can stably express humanα2A-adrenoceptor (α2A- AR). METHODS Recombinant plasmid of α2A- AR with hygromycin B (Hygro) resistance (pcDNA3.1/Hygro-HA-α2A-AR)was stably transfected into Chinese hamster ovary(CHO)cells which had expressed protein kinase A catalytic subunits(PKAcat) with labeling of enhanced green fluorescent protein(EGFP)by a Lipofectamine based method. A single positive clone expressingα2A-AR was selected through cultivation in the presence of 200 mg · L-1 hygromycin B followed by PKA redistribution assay. The transcriptional expression ofα2A-AR was detected by quantitative real-time PCR(qRT-PCR). Time-resolved fluorescence resonance energy transfer immunoassay was used to identify the function of inhibiting cAMP accumulation of α2A-AR. RESULTS The CHO-PKAcat-α2A-AR cell line No.7 exhibited stable response in PKA redistribution assay. qRT-PCR analysis demonstrated that the high expression ofα2A-AR in the cell line remained stable after a few generations compared with CHO-PKAcat-EGFP cells (P<0.01). The cAMP accumulation caused by forskolin was significantly inhibited by α2A-AR agonist in CHO-PKAcat-α2A-AR cells(P<0.01). CONCLUSION CHO-PKAcat-α2A-AR cell line is constructed successfully, which provides an effective model for drug screening and studies of mechanisms.

AIM: To evaluate the safety and tolerance of ibandronate in Chinese healthy volunteers. METHODS: The trial protocol was designed according to the Good Clinical Practice(GCP). After physical examination and laboratory tests were performed, 36 healthy volunteers were divided randomly into 6 dose groups, including 1 mg , 2 mg , 3 mg , 4 mg , 5 mg and 6 mg , with 6 subjects in each group(3 male and 3 female). Clinical symptoms, vital signs, routine blood tests, routine urine tests, hepatic function, renal function, blood electrolytes, electrocardiogram, and electroencephalogram were observed or examined before and after a single intravenous infusion of ibandronate. RESULTS: After single intravenous infusion doses of 1- 6 mg , the vital signs, clinical symptoms and laboratory tests were all in the normal range, but there were some slight ADRs concerned with the drug, such as hypophosphataemia, increased body temperature, perspiring,pain of bone or muscle and hypocalcaemia. But the ADRs were found vanishing in one or two weeks. CONCLUSION: Single intravenous infusion (up to 6 mg ) of domestic ibandronate in 36 chinese healthy volunteers is safe and tolerable.

OBJECTIVETo explore the neuro-protective effect and mechanism of qingkailing injection (QKL) against cerebral injury caused by E. coli-meningitis (CM).

METHODSThe CM model rabbits were treated by ampicillin with QKL as adjuvant. The leukocyte count and protein content in cerebral spinal fluid (CSF), the contents of water, sodium, potassium and calcium in cerebral tissues were measured before, 16 h and 26 h after Bacillus coli injection respectively. The expression of matrix metalloproteinase-9 (MMP-9) was determined at the same time.

RESULTSAdjunctive treatment with QKL can not only inhibit the increase of leukocyte cells, protein content in CSF, and water, sodium, calcium content in cerebral tissues, but also the decrease of potassium content revealed during simple antibiotic treatment. It also can decrease the expression of MMP-9 in cerebral tissues of rabbits with CM.

CONCLUSIONAs an adjunctive treatment, QKL can prevent transient inflammatory reaction and aggravation of brain injury in CM induced by simple antibiotic treatment, its mechanisms might relate with calcium antagonism and attenuation of MMP-9 expression in brain tissues.

Ampicillin ; therapeutic use ; Animals ; Anti-Bacterial Agents ; therapeutic use ; Brain ; metabolism ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Injections ; Male ; Matrix Metalloproteinase 9 ; biosynthesis ; Meningitis, Escherichia coli ; drug therapy ; Neuroprotective Agents ; therapeutic use ; Phytotherapy ; Rabbits

BACKGROUNDTo study the significance of the expression of HCV core protein in PBMC of patients with chronic hepatitis C and to evaluate the relationship between HCV core protein expression and clinical states.

METHODSIdentification of HCV protein antigen (Ag) in PBMC of 66 hepatitis C patients by immunohistochemical method and clinical status of the patients with HCV protein positive expression were investigated. In 27 out off 66 patients the HCV RNA and HCV Ag in PBMC were detected with RT-PCR and immunohistochemical method.

RESULTSThe HCV Ag (core+NS3) was identified in PBMC in 51 out of 66 patients (77.27%). It was also demonstrated that HCV core protein in nucleus showed strong expression and NS3 protein in cytoplasm showed weak expression. The expression of core protein in nucleus of PBMC were 35.29% in advanced chronic hepatitis patients, which was significantly higher than that from moderate cases (5.88%).

CONCLUSIONSThis study suggested that the expression of PMBC-HCV core protein may be related to the clinical state of the patients. The nuclear expression of HCV core protein in PBMC of patients with hepatitis C may be related to the persistence and activity of the chronic hepatitis C virus and play an important role in the pathogenesis of cirrhrosis and hepatocellular carcinoma.

Hepatitis C Antigens ; blood ; Hepatitis C, Chronic ; virology ; Humans ; Immunohistochemistry ; Leukocytes, Mononuclear ; metabolism ; RNA, Viral ; blood ; Viral Core Proteins ; blood