Objective To study the drug resistance mechanism and homologous relationship of four clinical strains of carbapen-em-resistant Klebsiella pneumoniae isolated from different sites of the same one patient.Methods Four carbapenem-resistant K .pneumoniae strains were isolated from blood,urine,sputum and pelvic drainage of a patient after cystectomy in clinical mi-crobiology lab of Xinhua Hospital in March 2012.① Modified Hodge test was used to detect the expression of carbapenemase.② PCR amplification assay and DNA sequencing were used to detect resistance-related genes.③ SDS-PAGE analysis was per-formed to analyze outer membrane porin components of the four carbapenem-resistant K .pneumoniae isolates.ERIC-PCR as-say was used to analyze the homology of these carbapenem-resistant isolates.Results Modified Hodge test demonstrated that all the four carbapenem-resistant K.pneumoniae expressed carbapenemase.KPC-2 gene was identified in all the four isolates by PCR test and DNA sequencing.SDS-PAGE analysis indicated an outer membrane porin alteration common to all the four iso-lates which is distinct from that of the strain sensitive to antibiotics.The identical DNA fingerprinting of the four strains was confirmed by ERIC-PCR analysis.Conclusions The antibiotic resistance mechanism of the four carbapenem resistant K.pneu-moniae associated with this case includes the expression of KPC-2 and altered outer membrane permeability induced by abnormal expression of outer membrane porin.These four strains belong to one common clonal group.

Objective To observe the effect of pulsed radiofrequency (PRF) on refractory infraorbital neuralgia. Methods From January, 2012 to December, 2014, 49 patients who received ineffective conservative therapy including medicine and nerve blockade underwent PRF treatment. Postoperative Numeric Rating Scale (NRS)=0 or 50%diminution of NRS was considered effective. NRS, effective rate, addition-al carbamazepine dosage and side effects were recorded on the first day, the first week, the second week, the first month, the third month, the sixth month, the first year, and the second year after operation. Results The effective rate were 67%, 67%, 65%, 59%and 51%on the first month, the third month, the sixth month, the first year, and the second year after operation, respectively. No serious side effect was observed, except that eight patients felt short-term (within one month) mild numbness. Conclusion PRF technique is safe and effective for refractory infraorbital neuralgia, and may become an alternative therapy.

The relationship between Bcl-2, Bax, Fas, caspase-3 and development of hemangioma and the molecular mechanism was investigated. By using immunohistochemical S-P method, proliferating cell nuclear antigen was detected. According to the classification of Mulliken in combination with PCNA expression, 27 cases were identified as proliferating hemangioma and 22 cases as involutive hemangioma. Five normal skin tissues around the tumor tissue served as controls. By using immunohistochemical technique, the expression of Bcl-2, Bax, Fax and Caspase-3 was detected. The cells expressing Bcl-2, Bax, Fax and cappase-3 were identified as hemangioma endothelia by immunohistochemical staining of VIII factor. The average absorbance (A) and average positive area rate of Bcl-2, Bax, Fas and caspase-3 expression were measured by using HPIAS-2000 imaging analysis system. The results showed that the expression of Bcl-2 in the endothelia of proliferating hemangioma was significantly higher that in involutive degenerative hemangioma endothelia and vascular endothelia of normal skin tissue (P < 0.01). The expression of Bax, Fas and Caspase-3 in the endothelia of involutive hemangioma was obviously higher than in the endothelia of proliferating hemangioma and normal skin tissue (P < 0.01). The expression of BAx and Fas in endothelia of proliferating hemangioma was higher than in those of normal skin tissue (P < 0.05). It was suggested that Bcl-2, Bax, Fas and caspase-3 might be involved in the development and involution of hemangioma. Bcl-2 could promote the growth of hemangioma by inhibiting apoptosis of endothelia. Bax, Fas and caspase-3 promote the switch of hemangioma from proliferation to involution by inducing the apoptosis of hemangioma endothelia.

Objective To explore the changes of plasma brain natriuretic peptide(BNP) and N-terminal pro-B-type natriuretic peptide (NT-proBNP)of sepsis combined with myocardial injury in newborns.Methods According to neonatal sepsis treatment program,45 cases of sepsis newborns in NICU of the Second Hospital in Lanzhou University from Jul.2007 to Jun.2008 were collected.According to the myocardial injury diagnostic criteria,45 cases neonatal sepsis were divided into myocardial injury group(n=22) and non-myocardial injury group(n=23).Myocardial injury group was also divided into congenital heart disease group and non-congenital heart disease group accor-ding to echocardiography.At the same time,30 healthy newborns were collected as healthy control group.Every newborns were tested the level of plasma BNP,NT-proBNP,creatine kinase-MB(CK-MB) and cardiac-troponin I (cTnI).Results There were significant difference between myocardial injury group,non-myocardial injury group and healthy control group in the levels of plasma BNP,NT-proBNP,CK-MB and cTnI,those in congenital heart disease group were higher than those in non-myocardial injury group and the healthy control group(Pa0.05).Conclusions BNP and NT-proBNP can be early used to diagnose myocardial injury and heart failure of neonatal sepsis associated with CK-MB and cTnI.In NICU,infants with sepsis should normally test BNP and NT-proBNP in order to early diagnose myocardial injury of neonatal sepsis.

Objective To observe the effect of radiofrequency thermocoagulation on patients with refractory supraorbital neuralgia Methods From February, 2007 to September, 2014, 36 patients with refractory supraorbital neuralgia following ineffective conservative ther-apy including medicine and nerve blockade underwent radiofrequency thermocoagulation. Numeric Rating Scale (NRS), Patient Satisfaction Scale (PSS), onset time, effective rate, recurrence rate, additional carbamazepine dosage and side effects were recorded before, and one day, one month, three months, six months, one year, two years after treatment. NRS=0 or 50%diminution was considered effectively. Results It worked two days on average (zero to seven) after treatment. The NRS score decreased (P<0.05) and the PSS score increased (P<0.05) com-pared with the data before treatment. The effective rate was 100%within six months. The recurrence rate was 11.1%within two years. Sec-ond radiofrequency thermocoagulation treatment still worked for the recurrent patients. No other side effect was observed, except permanent frontal numbness, short-time palpebral edema and ecchymosis. Conclusion Radiofrequency thermocoagulation is effective on refractory su-praorbital neuralgia, and the side effects are tolerable so it is an alternative choice for patients experiencing invalid conservative therapy.

The essential medium of B115 composed of 1% tryptone, 0.25% yeast extract and 0.5% sodium chloride was determined by using an orthogonal design. The orthogonal design was also employed in testing the optimum additions. It was composed of 0.1%(NH_(4))_(2)SO_(4),1.4%K_(2)HPO_(4), 0.6% KH_(2)PO_(4) and 0.1% (Na_(3)C_(6)H_(5)O_(7)). The yield of B115 cultured in optimum medium was compared with the one in essential medium. Statistic analysis showed that the growth of B115 was most significantly improved by adding K~(+)、NH~+_(4) and (Na_(3)C_(6)H_(5)O_(7)) to essential medium. The antibacterial effect of Bacillus subtilis strain B115 on pathogenic Aeromonas was studied. The results showed different antibacterial effects of B115 on different aeromonads. There were obvious antibacterial effects on BSK-10 and CL990920, while no effect on the growth of TL970424.

Objective To evaluate the effect of endoplasmic reticulum stress in trophocytes,in patients with intrahepatic cholestasis of pregnancy (ICP).Methods Sixty-one pregnant women who were hospitalized in Women's Hospital,School of Medicine,Zhejiang University from January to December 2015 were recruited.Thirty-one women who were diagnosed as ICP were defined as the ICP group and 30 healthy pregnant women were defined as the control group.The localization and expression intensity of glucose regulated protein 78 (GRP-78) in placental tissues were detected by immunohistochemistry technique.Electronic microscope was used to observe ultra-microstructure change of the endoplasmic reticulum in trophocytes and cell line Swan71.Reverse transcription (RT)-PCR and western blot were used to investigate the expression of GRP-78 mRNA and protein in Swan 71 cell.Results (1) GRP-78 protein was mainly expressed in the cytoplasm of cytotrophoblasts and syncytiotrophoblasts.The protein expression of GRP-78 in placentas of the ICP group (13.2±2.4) was significantly higher than that in the control group (7.8±1.3,P＜0.01).(2) The volume of endoplasmie reticulum did not increase and the microvilli developed well,with no swelling and no expansion of endoplasmic reticulum in the control group.In the ICP group,microvilli injury,endoplasmic reticulum edema were found;the volume of endoplasmic reticulum increased,with dilation,vacuolation and significant degranulation.After treated with 100 μmol/L cholyglycine for 24 hours,universal dilatation of the endoplasmic reticulum were seen in the Swan71 cells.(3) In Swan71 cells,cholylglycine displayed a concentration-dependent up-regulation on the expression of GRP-78.The expressions of GRP-78 mRNA in 0,25,50,100 μmol/L cholylglycine experimental group were 1.01±0.17,2.17±0.16,5.47±0.36,5.65 ± 0.82,respectively.The expression of GRP-78 protein in 0,25,50,100 μmol/L cholylglycine experimental group were 1.01±0.04,1.17±0.15,1.33±0.13,1.73±0.13,respectively.The expression of GRP-78 mRNA and protein in 100 and 50 μ mol/L cholylglycine experimental group were significantly higher than 0 μmol/L (all P＜0.01).Conclusion The obvious expansion of endoplasmic reticulum and the increased expression of GRP-78 in trophocytes indicated that endoplasmic reticulum stress of trophocytes may be involved in the pathogenesis of ICP.

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To investigate the relationship between sex hormone binding globulin (SHBG) and metabolic syndrome in elderly males in Shanghai,all the subjects (≥ 60 years old,male) underwent measurements of weight,height,waist and hip circumferences,and blood pressures,serum levels of fasting plasma glucose,total cholesterol,triglyceride,high density lipoprotein-cholesterol (HDL-C),and low density lipoprotein-cholesterol were determined (Hitachi,7600),while the levels of serum insulin,total testosterone,and SHBG were determined by using chemiluminescence methods.Free testosterone was calculated by using the Vermeulen equation.Metabolic syndrome was defined according to the criteria of the Chinese Diabetes Society (CDS 2004).The SHBG level in the metabolic syndrome group was significantly lower than that in non-metabolic syndrome group [(40.50 ± 26.16) nmol/L vs (47.80± 20.34) nmol/L,P＜0.01].With increasing number of metabolic syndrome components,the level of SHBG became lowered progressively.The subjects were divided into four subgroups according to SHBG Quartiles.From Quartile 1 to Quartile 4,body mass index,waist-hip ratio,diastolic blood pressure,fasting plasma glucose,triglyceride,fasting insulin,homeostasis model assessment for insulin resistance(HOMA-IR),free testosterone,free androgen index,and free testosterone percentage became progressively lowered,while age and HDL-C became raised (P＜0.05).SHBG was correlated significantly with age,body mass index,waist-hip ratio,diastolic blood pressure,fasting plasma glucose,HDL-C,and triglyceride.Age,HDL-C,and body mass index remained independently associated with SHBG in the multivariate regression analysis.In a logistic regression taking metabolic syndrome as the dependent variable,SHBG and HOMA-IR were included in the final model with statistical significance.Lowered SHBG is a risk factor of metabolic syndrome in elderly males.SHBG may be an independent predictor of metabolic syndrome,but the mechanism of how SHBG is involved in the metabolic syndrome needs to be further studied.

s were 0.573±0.016, 0.707±0.008 and 0.711±0.013). Conclusions CXCL10 can express stablely in MCF-7 cell lines, which resulted in down-regulation of expression of VEGF and STAT3 gene. CXCL10 played an important role in anti-tumor effect.