Objective To establish the standardization process of acid-base balance analysis, analyze cases of acid-base balance disorder with the aid of acid-base balance coordinate graph.Methods The acid-base balance theory were reviewed systematically on recent research progress, and the important concepts, definitions, formulas, parameters, regularity and inference in the analysis of acid-base balance were studied. The analysis of acid-base balance disordered processes and steps were figured. The application of acid-base balance coordinate graph in the cases was introduced.Results The method of four parameters-four steps analysis was put forward to analyze the acid-base balance disorders completely. Four parameters included pH, arterial partial pressure of carbon dioxide (PaCO2), HCO3- and anion gap (AG). Four steps were outlined by following aspects: ① according to the pH, PaCO2 and HCO3-, the primary or main types of acid-base balance disorder was determined; ② primary or main types of acid-base disorder were used to choose the appropriate compensation formula and to determine the presence of double mixed acid-base balance disorder; ③ the primary acid-base balance disorders were divided into two parts: respiratory acidosis or respiratory alkalosis, at the same time, the potential HCO3- should be calculated, the measured HCO3- should be replaced with potential HCO3-, to determine whether there were three mixed acid-base disorders; ④ based on the above analysis the data judged as the simple AG increased-metabolic acidosis was needed to be further analyzed. The ratio of ΔAG↑/ΔHCO3-↓ was also needed to be calculated, to determine whether there was normal AG metabolic acidosis or metabolic alkalosis. In the clinical practice, PaCO2 (as the abscissa) and HCO3- (as the ordinate) were used to establish a rectangular coordinate system, through origin (0, 0) and coordinate point (40, 24) could be a straight line, and all points on the straight line pH were equal to 7.40. The acid-base balance coordinate graph could be divided into seven areas by three straight lines [namely pH = 7.40 isoline, PaCO2 = 40 mmHg (1 mmHg = 0.133 kPa) line and HCO3- = 24 mmol/L line]: main respiratory alkalosis area, main metabolic alkalosis area, respiratory+ metabolic alkalosis area, main respiratory acidosis area, main metabolic acidosis area, respiratory+ metabolic acidosis area and normal area. It was easier to determine the type of acid-base balance disorders by identifying the location of the (PaCO2, HCO3-) or (PaCO2, potential HCO3-) point on the acid-base balance coordinate graph.Conclusions Four parameters-four steps method is systematic and comprehensive. At the same time, by using the acid-base balance coordinate graph, it is simpler to estimate the types of acid-base balance disorders. It is worthy of popularizing and generalizing.

Objective: To research three-dimensional computed tomography angiography (3D-CTA) combined three-dimensional digital subtraction angiography (3D-DSA) in the diagnosis of intracranial aneurysms. Methods: 3D-CTA and 3D-DSA were performed in 27 cases with Highly suspected in patients with intracranial aneurysms, And compared with the final outcome of surgery. To evaluate 3D-CTA and 3D-DSA in the detection rate of aneurysms, three-dimensional shape and spatial relations ability to display. Research 3D-CTA combined 3D-DSA in the diagnosis of intracranial aneurysms. Results: Twenty-seven cases of suspected patients, 25 cases of surgical treatment of confirmed aneurysms, 28 aneurysms found. 3D-CTA and 3D-DSA rate of aneurysms was no significant difference. 3D-CTA of the aneurysm can be accurately detected clearly shows its three-dimensional shape and spatial relationship with the surrounding skull, 3D-DSA for aneurysm neck, the tumor and the aneurysm and parent artery relationship shows clear and accurate. Conclusion: Imaging of intracranial aneurysm diagnosis, 3D-CTA and 3D-DSA have their respective advantages, combined two methods can improve the detection rate of aneurysms. 3D-CTA combined 3D-DSA have support of design for operation.

Objective To identify the effects of virus specific amino acids in the fusion active domains of paramyxovirus fusion proteins on the specific membrane fusion. Methods Site-directed mutagenesis was used to obtain mutants in the identified fusion active domains of Newcastle disease virus (NDV) fusion protein (F) and human parainfluenza virus (hPIV) fusion protein (F). All the mutant F genes were co-expressed with their homol-ogous or heterogenous hemagglutinin-neuraminidase (HN) genes in eukaryocytes. The fusion functions of mutants were assayed by Giemsa staining and reporter gene method. The expression efficiencies of mutants were assayed by fluorescence-activated cell sorter (FACS). Results In the NDV F mutants, N150D-L152D had 46.31% fusion activity of wide type. The fusion activities of N257D-N258D-Q259E, G271D-N272D and Q279E-Q281E almost disappeared, and they had only 1.25%, 3.14% and 2.23% of fusion activities, respectively, compared with wide type. N296D-N297D had 97.68% fusion activity of wide type. In the hPIV F mutants, D143A-E145A had 32.63% fusion activity of wide type. The fusion activity of E223Q-K224A almost disappeared, and it had only 1.91% fusion activity of wide type. K263A-R265A, D268A-D270A and R475A-R476A had 14.63%, 19.52% and 28.95% of fusion activities respectively compared with wild type. The analysis of FACS indicated that proteins of NDV F N257D-N258D-Q259E, G271D-N272D, Q279E-Q281E and hPIV F E223Q-K224A were not expressed on the cell surface, while proteins of the rest mutants were expressed nearly as the same as the wide types. Con-clusion As to NDV F, the amino acids of N257, N258, Q259, G271, N272, Q279 and Q281 were significant to the specific membrane fusion, and N150 and L152 were also important, but N296 and N297 were not. For hPIV F, the amino acids of E223 and K224 were significant to the specific membrane fusion, and D143, E145, K263, 11265, D268, D270, R475 and R476 were also important.

Objective:To clone human interleukin-33(hIL-33)and express it in E.coli efficiently.Methods:The primers were synthesized according to the hlL-33 cDNA sequence in GeneBank.The hIL-33 was amplified by RT-PCR from human fibroblast cell line(L929),the PCR product was inserted into pUC19 vector.The IL-33 cDNA confirmed by sequencing was inserted into expressing vector PQE30 and expressed in E.coli M15 strain.IL-33 protein expression was induced by IPTG and purified by Ni-NTA affinity chromatography.The recombinant IL-33 was identified by Immunoblot and its biological activity was analyzed.Results:DNA sequencing confirmed that the cloned cDNA was identical to the published sequence of hIL-33.The recombinant plasmid PQE/hIL-33 was transformed into M15.An expected 18KD protein of hIL-33 found mainly in the induced host strains about 25% of total bacteria lysis by SDS-PAGE and coomassie blue staining.The 18 KD protein could be recognized by anti-IL33 antibody in western blot.The recombinant protein was purified to more than 95% of total protein and induces the production of IL-4 and IL-5 in human peripheral blood mononuclear cells.Conclusion:We have successfully expressed hIL-33 protein in E.coli and the expressed product has IL-33 specific bioactivity.

Objective:To investigate the predictive value of P-selectin and D-dimer in the early diagnosis of portal vein thrombosis(PVT) in patients submitted to portal hypertension operation. Method:Eighty-two patients were assigned into two groups(PVT group and non-PVT group)after the operative treatment of portal hypertension.The levels of P-selectin and D-dimer were detected dynamically in the patients of two groups.Results:The preoperative levels of P-selectin and D-dimer in the PVT group were higher than those in the non-PVT group respectively(P

BACKGROUND:Cerebrovascular stent may destroy the vessel walls,which can lead to vascular restenosis.There are different versions about the safety,pathologic pharmacology reasons and clinical effect.OBJECTIVE:To evaluate the safety and short-term effect of endovascular stent implantation for symptomatic artery stenosis.METHODS:Totally 20 patients with total 22 lesions diagnosed symptomatic artery stenosis were treated with endovascular stenting.The vascular stenotic lesions involved middle cerebral artery in 6 cases,internal carotid artery in 6 cases,vertebral artery in 4 cases,basilar artery in 3 cases and vertebro-basilar artery in 3 cases.The length of vascular stenotic lesions was 3-10 mm with the average of 7 mm.Both balloon and self-expandable stents were used in 12 cases with embolus protection device.RESULTS AND CONCLUSION:All of the 22 stents in 20 patients were placed successfully in one time.After stent implantation,the vascular angiography showed that the vessels were reformed obviously with the degree of stenosis no more than 20%.The perfusion in cerebrum was improved in parenchymal phase and the symptom was also improved clearly.At 6-24 months follow-up,20 patients never had cerebral ischemia.With follow-up for 12-24 months in 7 patients,digital subtraction angiography displayed that intima hyperplasia was occurred in stent in one patient with no symptom.The stenotic vascular were unobstructed and no intima hyperplasia by transcranial Doppler examinations.The results demonstrated that endovascular stent implantation is a safe and effective treatment for intracranial symptomatic artery stenosis,while its long-term effect needs further study.

The construction of biosafety laboratory was highly concerned these years because of the spread of high pathogenic diseases including SARS and avian influenza. In the past several years, the construction of many BSL-2/3 laboratories played an important role in preventing the dissemination of pathogens involved in the research. Meanwhile, serious problems were also raised because some project designers or constructors were not clear with the compulsory safety equipments in these laboratories so that the requirement of zero-leakage could not be achieved. The compulsory safety equipments emerging and disposing solid, liquid and gas offal in the biosafety laboratory were reviewed. The R&D of special equipments in biosafety laboratory in China was also introduced.

Objective To evaluate the international blood smear review criteria for improvement using Beckman-Coulter hematology analyzers and find out proper slide review criteria suitable for Chinese population. Methods 3 600 random-selected blood samples were tested in three hospitals using MAXM, GENS and LH750 5-diff automated analyzers and a manual differential with a smear review was performed on all samples in the study. True positive rate, true negative rate, false positive rate and false negative rate were calculated according to the international blood smear review criteria. We have set up smear review criteria for Chinese population by analyzing false positive and false negative cases according to Chinese clinical conditions. Another 240 blood samples were tested in three hospitals using the same analyzers to verify the new slide review rules. Results According to international blood smear criteria, the true positive rate was 4.9%, false positive rate was 24.2%, true negative rate was 70.4% and false negative rate was 0.5%. The international smear review criteria were modified into 23 criteria by analysis of above statistical data. In addition, we added four WBC differential ratio rules in positive smear criteria. After modification, the true positive rate was 9.9% (355/3 600) ,false positive rate was 17.1% (617/3 600), true negative rate was 71.2 % (2 563/3 600) and false negative rate was 1.8% (65/3 600). No blast cell was missed using both smear review criteria. A little higher false negative rate after modification was caused by supplemented differential ratio roles in positive smear criteria. Verification results were satisfactory;The tree positive rate was 13.7% (33/240) ;false positive rate was 15.8% (38/240) ;false negative rate was 2.5% (6/240) and true negative rate was 68.0% (163/240). Conclusions Although the smear review criteria suggested by the International Consensus Group is clinically important, the false positive rate increases when they are used in Chinese population. The modified slide review criteria used on Beckman-Coulter hematology analyzers in this study are more suitable for Chinese laboratories.

Objective To observe the clinical efficacy of metronidazole tablets combined with sophora gel in treatment of bacterial vaginitis ( BV) . Methods Eighty-seven cases of BV patients were randomly divided into the study group(45cases) and the control group(42cases).The control group was given metronidazole tablets (400 mg, two times a day), oral for seven days, while the study group was given sophora gel (vaginal implantation) on the basis of control group.Seven days as a course of treatment.Pared the leukocyte esterase test positive rate in vaginal fluid and the recurrence rate within a year of the two groups before and after treatment.ResuIts After 7 days’treatment, the positive rate of leukocyte esterase (LE) in two groups were all significantly declined, which was more significant in the study group (P<0.05);the total effective rate of the study group was 95.6%, which was significantly higher than that of the control group 78.6%(P<0.05).The recurrence rate within a year of the study group was 6.7%, which was significantly lower than that of the control group 26.2%( P<0.05 ) .ConcIusion The therapy of metronidazole tablets combined with sophora gel in treating BV can significantly decline the LE positive rate and recurrence rate, and improve the clinical curative efficacy as well.