ObjectiveTo study the regularity of the changes and the role of serum ionized Calci um in the pathogenesis of hemorrhagic fever with renal syndrome(HFRS). MethodsSerum was detected at different stages of HFRS by selective electrode, and analized the relationship between serum with blood urea Nitrogen and creatinine. ResultsSerum was much lower (P＜0.05) during hypotension shock stage and diuretic stage, and significantly lower(P＜0.01) during oliguria stage. The decrease of serum level was correlated to the state of illness aggravated. There is a negative relativity between serum and blood urea Nitrogen and creatinine (P＜0.01) in the patients. ConclusionSerum levels in patients may be related to the severity of the disease and renal function.

Objective To observe the effect of ketamine preemptive analgesia on postoperative pain and recovery in laparoscopic cholecystectomy(LC). Methods Eighty LC patients under general anesthesia with propofol and remifentany were randomly divided into two groups of PR and PRK with 40 cases each. The patients in group PRK were given additional low-dose ketamine 0.3 mg/kg before skin incision. Recovery from anesthesia was evaluated at 30 min, 1 h and 12 h after surgery by SS and VRS scoring. Results There were no significant differences in the time for regaining spontaneous breathing,opening eyes and extubation between the two groups. But recovery of anesthesia was with less restlessness and VRS pain scores, better sedation in group PRK than those in group PR (P< 0.05). Conclusion Preemptive intravenous analgesia with low-dose ketamine has the advantage of quicker recovery from anesthesia with less incidence of restlessness and pain in LC patients.

Objective To analyze the direct interaction between mmu-miR-30b and mouse FoxO3 (mFoxO3) mRNA.Methods Three target gene fragments, which were respectively 402, 123 and 299 bp in length, were amplified from mouse cDNA by PCR using specific primers and site-direct mutant primers.A complete mutant fragment was obtained by joining together the 123 and 299 bp fragments.Recombinant plasmids, pmirGLO-mFoxO3 and pmirGLO-mFoxO3 mt, were constructed through inserting wild and mutant fragments of mFoxO3 into pmirGLO vector, respectively.HEK 293T cells were respectively co-transfected with the constructed recombinant plasmids and mmu-miR-30b/mmu-miR-30b inhibitor.Dual-luciferase reporter assay system was used to determine the Firefly-Renilla luciferase activity in different groups.Western blot assay was performed to evaluate the regulatory effect of mmu-miR-30b on mFoxO3 expression.ResultsRestriction enzyme analysis and gene sequencing showed that the two recombinant plasmids, pmirGLO-mFoxO3 and pmirGLO-mFoxO3 mt, were constructed successfully.The activity of Firefly-Renilla luciferase in HEK 293T cells transfected with pmirGLO, pmirGLO+mmu-miR-30b, pmirGLO+mmu-miR-30b inhibitor, pmirGLO-mFoxO3+mmu-miR-30b, pmirGLO-mFoxO3+mmu-miR-30b+mmu-miR-30b inhibitor, or pmirGLO-mFoxO3 mt+mmu-miR-30b was 13.18±0.97, 14.35±0.99, 12.46±1.20, 9.55±1.11, 13.71±0.89 and 10.99±0.92, respectively.Compared with pmirGLO+mmu-miR-30b group, the luciferase activity was significantly decreased in pmirGLO-mFoxO3+mmu-miR-30b group (P<0.05), but showed no significant change in pmirGLO-mFoxO3 mt+mmu-miR-30b group (P>0.05).In addition, the suppressed Firefly-Renilla luciferase activity in pmirGLO-mFoxO3+mmu-miR-30b group was restored by mmu-miR-30b inhibitor treatment (P<0.05).Enhancing the expression of mmu-miR-30b could markedly inhibit the expression of mFoxO3 at protien level (P<0.05), and that could be significantly attenuated by mmu-miR-30b inhibitor treatmeat (P<0.05).Conclusion mFoxO3 mRNA is a novel target gene of mmu-miR-30b.There is a direct interaction between mmu-miR-30b and mFoxO3 mRNA.

Objective To explore the methods of culture and identification of human placental microvascular endothelial cells(HPMVEC) in vitro,aiming to construct the polar model of human placental barrier.Methods By proteolytic enzymes,the tissular and cellular suspensions were collected from the chorionic villi of pregnant and centrifugated over a preformed discontinuous Percoll gradient.The highly purified human placental microvascular endothelial cells were obtained and cultured in HG-DMEM with fetal calf serum and identified by immunocytochemistry(SP methods) and transmission electron microscope.Results The highly purified human microvascular endothelial cells were cultured in vitro and passaged for three to five generations.Factor Ⅷ-related antigen and CD34 were both positive by immunocytochemistry.Weibel-Palade body(W-P body) was observed by transmission electron microscope.Conclusion Human placental microvascular endothelial cells can be isolated and cultured successfully in vitro by our method.

Objective:To study the distribution of the literature and the trend of international research of proliferative vitreoretinopathy (PVR) in 10 years from 2009 to 2018.Methods:Using Web of Science database of the American Institute of Scientific Information as the data source to collect literature on proliferative vitreoretinopathy published from 2009 to 2018 and a statistical analysis on the distribution of the publication time, nation and funding agency was made to figure out the institution, periodical, author with highest citations respectively among the collected literature.Meanwhile, the bibliometric analysis software Citespace and VOSviewer were employed to carry out the distribution analysis and cluster analysis of keywords as well as the co-citation cluster analysis.Results:A total of 905 articles related to PVR were included, mainly in English, most of which were written by American authors, and the number of literature published by Chinese authors was ranked the second.Among all the article funding institutions, the National Natural Science Foundation of China ranked NO.1 with the most funded articles.Citations increased year by year since 2009 and increased obviously since 2013.The journal with the highest citations was Investigative Ophthalmology & Visual Science, and the research institution with the highest citations was Harvard University.The statistical analysis of the citation quantity also showed that the top five authors with the highest citations were all from Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School.The results of cluster analysis showed that there were 41 high-frequency subject terms, and they were mainly grouped in 4 categories, which were etiology, risk factors, molecular mechanism, treatment and management of PVR.Keywords network visualization and overlay visualization both showed that etiology and risk factors were the focus of current research, while epithelial mesenchymal transition, transforming growth factor-β and fibrosis were new research fields in recent years.The results of citation cluster analysis showed there were 12 co-citation clusters. Conclusions:The literature of PVR shows an upward trend, and the researches focus on the molecular mechanism, prevention, and neuroprotection of PVR, which may be the future PVR research direction.

【Objective】To investigate the expression of nitric oxide synthase 3 (NOS3) and vascular endothelial growth factor (VEGF) in hepatocellular carcinoma (HCC).【Methods】The expression of NOS3 and VEGF were detected by immunohistochemistry in 51 cases of HCC.【Results】The positive of NOS3 and VEGF in HCC were 53% and 63%.The expression rate of NOS3 in the recurrent group was significantly higher than that in the non-recurrent group (P<0.01).The expression rates of VEGF in the group with carcinoma embolus of portal vein and the recurrent group were significantly higher than those in the group without carcinoma embolus and non-recurrent group (P<0.05).The expression of NOS3 was closely related with the expression of VEGF positively in HCC.【Conclusions】NOS3 and VEGF are related with the biological behavior of HCC closely.

Abstract-Eleven mouse monoclonal antibodies (McAbS)specific for human alpha-fetopr-otein(AFP) have been produced by the hybridoma technique. Immunodiffusion was us-ed for determining the immunoglobulin class and subclass of the McAbS. Seven of them are IgG; two are IgM. At least 4 different antigenic determinants on human AFPcan be recognized by using the McAbS and cross-two-site sandwich solid radioimmun-oassay. One of the McAbS can specially react with one of the fragments of AFP dig-ested by pepsin. In addition, the cross-reactivity among the AFPs of different mam-malian species was also tested by using the McAbS.

Objective: To study the super high-dose chemotherapy in drug-resistance Ⅲ B osteasarcoma with autologous bone marrow transplantation. Methods: The chemotherapy was carried out in 4 patients with drug-resistance Ⅲ B osteosarcoma. The autologous bone marrow was harvested before chemotherapy and reinfused after chemotherapy when the serum concentration of methotrexate reduced to 1.0?10~(-7) mol/L. The chemotherapy included ,methotrexate 500 mg/kg , adriamycin 25 - 75 mg/m~2 and vincristine 1.5 mg/m~2 with low dose citrovorum factor rescue. Results: Four patients tolerated the chemotherapy well and felt better than before. The pulmonary metastases had various degrees of reduction in 2 patient and calcified in 1 patient, but progressed in 1 patient. Conclusion: Super high-dose chemotherapy has good response in drug-resistance ⅢB osteosarcoma and is safe with autologous bone marrow transplantation. This regimen is worth to be further studied and be used in ⅡB osteosarcoma.

Objective To determine the applicability of locally continuo u s application of alendronate for the prevention of prosthesis aseptic loosening. Methods Using the method of in vitro cell culture to detect the influence of a lendronate with different concentrations(1?10－11mol/L,1?10－9mol/L,1?10－7mo l/L,1?10－6mol/L,1?10－5mol/L) on the proliferation, secretion and osteogenesi s of human osteoblasts as well as the effect of the motility, respiration, phago cytosis and bactericidal ability. Results Higher concentration of alendronate(1 ?10－6mol/L,1?10－5mol/L) improved local antiinfection ability, but inhibited osteogenesis (1?10－5mol/L). Lower concentration(1?10－11mol/L,1?10－9mol/L,1 ?10－7mol/L) inhibited the bactericidal effect of neutrophils, but stimulated o steogenesis(1?10－11mol/L,1?10－9mol/L). Conclusion Locally continuously usin g alendronate around prosthesis to prevent and treat aseptic loosening must be h andled with discretion. Its concentration should not be below 1?10－6mol/L in the early period after arthroplasty operation, but the concentration should be 1?10－11mol/L or 1?10－9mol/L in the late period.

OBJECTIVE:To study the preparation technic and determination of the content and dissolubility of the fluconazole-?-cyclodextrin inclusion complex(FCIC) METHODS:The FCIC was prepared by coprecipitation method and ultr_asonic method The content and percentage of the accumulated dissolubility of FCIC were determined by UV-spectrophotometry RESULTS:Fluconazole and ?-cyclodextrin could form inclusion complex through either preparation technology The contents of fluconazole in the complex were(11 34?0 77)% and(11 23?0 24)% respectively The dissolubility met the standard in CP CONCLUSION:Stable complex could be formed between fluconazole and ?-cyclodextrin