Objective To analyze the 23S rRNA gene partial sequences of common bacteria, and establish molecular biologic techniques to identify bacteria by the difference of gene sequences. Methods Analyzing the sequences of variable region of bacterial 23S rRNA genes, primers and oligonucleotide probes were designed accordingly. Thereafter, bacteria were identified by PCR gel electrophoresis and PCR reverse hybridization. Results There exists significant sequence difference between Gram negative bacteria and Gram positive bacteria and it could be used to differentiate these 2 kinds of bacteria quickly with PCR gel electrophoresis. Meanwhile, sequence variety in different species of bacteria was also observed and PCR reverse hybridization could be used to identify different bacterial species further.Conclusions There exist significant sequence differences among 23S rRNA genes in different common bacteria. By the sequence differences, a specific, sensitive and rapid molecular biologic techniques could be established to quickly identify the pathogens of bacterial infections.

Objective To explore the expression of sterol regulatory element binding protein 1C (SREBP-1C) and glucose-regulated protein 94(GRP-94)in hyperhomocysteinmia and to evaluate the effects of endoplasmic reticulum stress proteins on hepatocytes lipids metabolism. Methods After hyperhomocysteinmia C57BL/6 mice model being induced by high methionine diet, TGE and CHO of Hepatocytes were determined, and the expression of SREBP-1C and GRP-94 was assessed by RT-PCR and Western blot. All data were compared to those in control group′s. Results The level of plasmic homocysteine(Hcy) and hepatocytes TGE or CHO of high methionine diet mice at different time point significantly ascended(P

Fluorescence probe Fluo-3AM can be luminary in confocal microscope after combining Ca2+,So nomadic Ca2+ in cytoplasm can be measured and tested by this way.Some confocal laser scan images are obtained after calcium in muscle fibers signed with F1uo-3AM. Then mapping function is used to convert the gray level image into colorful image by pseudo-color enhancement theory in digital image.Compared to the original gray level image,the processed pseudo-color image could be more easily distinguished.So the methods had a great value in Ca2+ detection fields.

Objective To explore effect of CYP3A4 gene polymorphism and genetic testing on the efficacy of fentanyl in patients with extensive burns.Methods A prospective randomized controlled study, 149 patients with extensive burns were picked from Feburary to July in 2015.Patients were randomly divided into matched group ( n=56 ) and experimental group ( n=93 ) . The matched group was treated with fentanyl 5 g/kg to finish implementation of anesthesia induction.Depends on the gene detection results, the patients in experimental group were treated with 6 g/kg, 5 g/kg or 4 g/kg fentanyl.The VAS score at different time after operation in the experimental group and the control group were compared , and the dosage of fentanyl was observed.Patients whose VAS score was greater than 7 should treated with intramuscular injection of pethidine 100 mg, recorded all patients with pethidine,s additional cases and the times.At the same time, the adverse reactions of the two groups were recorded before and after hospital discharge. ResuIts The score of VAS was not significantly different after wake up immediatly after the surgery , and the score of VAS was significantly lower in the experimental group patients in 6h,12h after the surgery (P<0.05).The dosage of fentanyl in the experimental group and the number of the cases and the times were significantly decreased (P<0.05).The adverse reactions after the surgery such as nausea, vomiting, itching, drowsiness, and adverse reactions were significantly lower in the experimental group (P<0.05).ConcIusion CYP3A4 gene detection in patients with large area of burn in the individual drug use is important, because of patients’ genotype adjusting fentanyl dosagecan, it enhances significantly the analgesic effect, reduces the amount of drug use, and effectively reduces the adverse reactions.

Objective To compare the uniformity between spine MRI and 18F-FDG PET/CT in detection of spinal metastatic tumor, and to analyze the reasons for the discrepancy between them.Methods Data of 42 malignant tumor patients underwent spine MRI and 18F-FDG PET/CT were analyzed retrospectively.And the two tests were underwent within 30 days.The author censused and analyzed the number of centrums with metastatic tumor detected by spine MRI and 18F-FDG PET/CT,and supposed that 18F-FDG PET/CT was gold standard,to evaluate the sensitivity and specificity of spine MRI.The inconsistent between two tests was analysed.Results The study included 509 centrums of 42 patients.123 positive centrums (24.2%) were detected on spine MRI,and 114 positive centrums (22.4%) on 18F-FDG PET/CT(P=0.22>0.05).The uniformity between two tests was good (Kappa=0.76).Supposing that 18F-FDG PET/CT was gold standard,the sensitivity,specificity,positive predictive value, negative predictive value,false negative rate and false positive rate of spine MRI was 85.1%(97/114),93.4%(369/395),85.1%(97/114),93.4% (369/395),14.9%(17/114) and 6.6%(26/395) respectively.43 centrums of 14 patients were diagnosed inconsistently by spine MRI and 18F-FDG PET/CT.The main reasons for the inconformity were as following: types of primary tumors, insensitivity of spine MRI for diffuse scattered spinal metastatic tumors, the existence of misdiagnosis on spine MRI, misdiagnosis of spine MRI for metastatic lesions located in the vertebral appendix and paramedian vertebral body,misdiagnosis of 18F-FDG PET/CT for metastatic lesions located in the sacral centrum, existence of false positive on 18F-FDG PET/CT and so on.Conclusion The uniformity between spine MRI and 18F-FDG PET/CT is good.The inconformity still existes in some cases.Carefully comparative analysis of them contributes to find metastatic lesions.

AIM:To explore the effect of homocysteine (Hcy)-mediated endoplasmic reticulum stress on lipid metabolism in hepatocytes.METHODS: HepG2 cells used in the study were treated with 5 mmol/L Hcy. The concentrations of Hcy, triglycerides (TGE) and cholesterol (CHO) in the cells were measured. In high methionine diet-induced hyperhomocysteinemia C57BL/6 mice, the concentrations of TGE and CHO in hepatocytes were analyzed. The mRNAs and proteins expressions of glucose-regulated protein 78 (GRP-78) and sterol regulatory element binding protein (SREBP-1) were also assessed.RESULTS: The concentrations of Hcy and lipids (TGE, CHO) in HepG2 cells at different time point were elevated after treated with 5 mmol/L Hcy (P0.05,vs 0 week). The mRNAs and proteins expressions of GRP-78 and SREBP-1 in mice at different time point after high methionine diet were higher than that at 0 week (P

This paper gives the techniques and steps of creating remote control instruments system using Windows API functions,and explains its hardware and software structures.The whole system is proved to be stable and reliable by experiment.It can provide a new experimental method for biomedical research,which enable the construction of multifunctional experiment platform.

Objective To characterize clinical feature, frequency of isolation and antimicrobial susceptibility of pathogens isolated from patients with nosocomial bloodstream infections in Huashan Hospital, Fudan University from 1995 to 2004. Methods The clinical data of all patients who were diagnosed with nosocomial bloodstream infections based on national diagnostic criteria of nosocomial bloodstream infections were retrospectively analyzed. The pathogens were routinely isolated and identified. Susceptibilities against antimicrobial agents were determined by Kirby-Bauer methods and analyzed by WHONET 5.0 software. Results During the 10-year study period, a total of 395 patients were diagnosed with nosocomid bloodstream infection with 435 strains isolated from blood specimen.Gram positive bacteria, Gram negative bacilli and fungi, accounted for 47.4%, 45.1 % and 7.6%,respectively. Coagulase-negative Staphylococci (21.4%), S. aures (17.9%), E.coli (13.6%), K. pneumoniae (10.8%), Candidaspp (7.4%), Enterococci (6.0%), Pseudomonasspp (6.0%) and Acinetobacter spp (3.7%) were frequently identified isolates. S. aures and coagulase-negative Staphylococci resistant to methicillin were 62.8% and 87.1%, respectively. The susceptibilities of cefotaxime and ceftazidime against E. coli and K. preumonine were 46%-78% and 27.7%-40.4%, respectively. Conclusions The Gram positive cocci are slightly more prevalent than Gram negative bacilli in nosocomial bloodstream infections and resistance to the first line antibiotics is common among all pathogens isolated. Candida spp is the fifth leading cause of nosocomial bloodstream infections.

Objective To understand the resistance mechanism and clinical feature of linezolid-resistant S.capitis isolated from blood samples.Methods Antimicrobial susceptibility testing was carried out to determine the susceptibility of clinical strains.PCR and sequencing analysis were used to analyze cfr gene and 23S rRNA mutation,which were associated with linezolid resistance.Patterns of pulsed-field gel electrophoresis (PFGE) were analyzed in combination with clinical data to understand the clinical feature of S.capitis strains.Results Five linezolid-resistant S.capitis strains were isolated from blood samples of 3 patients.These strains were resistant not only to linezolid,but also to most of the commonly used antimicrobial agents except glycopeptides,rifampin,and trimethoprim-sulfamethoxazole.Mutation was identified in 23S rRNA genes of all the five strains and cfr gene was found in four of the five strains.PFGE typing showed the same type,which supported the homology of the 5 strains.Three patients had deep vein indwelling catheter and two of them were treated with linezolid.Conclusions Linezolid-resistant S.capitis isolates showed the phenotype of resistance to multiple antimicrobial agents.Linezolid resistance may be mediated by cfr gene and 23S rRNA mutations in S.capitis.Long-term use of deep vein indwelling catheter and linezolid treatment may increase the risk of linezolid-resistant S.capitis infection.

AIM: To inhibit the expression of tyrosinase gene in murine B16 melanoma cells by antisense nucleotide. METHODS: The antisense recombinant pcDNA3.1(-)-tyr was constructed and was used to infect murine B16 melanoma cells for expression of tyr antisense nucleotide. The effect of antisense nucleotide of tyr on the expression of tyr gene was detected by determination of the activity of tyrosinase and of the production of melanin, Dopa staining and electronic microscope. RESULTS: The tyr antisense recombinant was successfully constructed and injected into murine B16 melanoma cell. The activity of tyrosinase in B16 cells infected with pcDNA3.1 (-)-tyr decreased to 0.0498?0.0036, compared to the tyrosinase activity of 0.0916?0.0132 in the control cells without treatment (P