Objective To investigate the prevalence of 16S rRNA methylase genes, armA and rmtB, which mediate high level aminoglycoside resistance, in gram-negative bacteria isolated from 2 hospitals in Dalian and study the mechanism of aminoglycoside resistance.Methods A total of 134 amikacin-resistant clinical isolates of gram-negative bacteria were collected. Two 16S rRNA methylase genes, armA and rmtB, were identified by PCR-based assays. PCR products were extracted for DNA sequencing analysis. armA and rmtB gene mapping were conducted by plasmid extraction, conjugation and transformation. The MICs of amikacin, gentamicin and tobramycin were determined for the positive isolates, transconjugants and the resultant strain of transformation using agar dilution technique. Results Overall armA was identified in 21 strains of Acinetobacter baumannii, rmtB in 5 strains of Escherichia coli and 5 strains of Klebsiella pneumoniae. Plasmid extraction and conjugation experiments were only successful for rmtB-positive isolates. Transconjugant and DH5a (pMDarmA) exhibited high-level resistance to aminoglycosides.Conclusions The 16S rRNA methylase genes, armA and rmtB are identified in Dalian. armA gene is identified in A. baumannii. rmtB gene is located on the plasmid of E. coli and K. pneumoniae. armA and rmtB can induce high-level resistance to aminoglycosides.

Objective To investigate the types of commonly used anti-hypertensive drugs and hypoglycemic drugs and treatment schemes to provide a basis for rationally treating hypertension complicating type 2 diabetes mellitus(T2DM).Methods The medication situation in 191 patients with hypertension complicating T2DM in our hospital were analyzed and investigated.The use of anti-hypertensive drugs and hypoglycemic drugs and treatment scheme were statistically analyzed.Results In 191 patients with hypertension complicating T2DM,the single drug use in the anti-hypertensive scheme accounted for 9.42 ％,two-drug combined use accounted for 64.91％,three-drug combined use for 17.8 ％ and four-drug combined use for 7.85 ％.The medication of ARB+CCB had the highest use frequency;the anti-hypertensive total effective rate was 96.86 ％.In the hypoglycemic schemes,the insulin use accounted for 30.37％,and the use frequency of metformin+ gliclazide was highest,the hypoglycemic total effective rate was 98.43％.Conclusion The medication for the patients suffering from hypertension complicating T2DM is relatively reasonable,and worth of being popularized and applied in clinical practice.

Objective To detect the expression of cyclooxygenase(COX) in human gastric cancer cell lines and its subcellular location of the isoforms. Methods Immunohistochemistry、RTPCR combined with laser scanning cofocal microscopy (LSCM) were used to investigate expression and distribution of COX. Results Positive staining of COX2 and COX1 protein was seen in human gastric cancer cell line MKN45、SGC7901 and AGS. However, the COX2 staining was absent and COX1 staining was weak in MGC803; though their mRNA in all the four cell lines. When compared to COX1,COX2 showed a stronger signal at both protein and mRNA levels of the gastric cancer cell lines, which was confirmed by double labeling and LSCM. The quantitative analysis of fluorescein intensity indicated the pixel intensity peak of COX2 reached 50～70, while COX1 only 10. LSCM also showed that COX2 was both cytoplasmic and nuclear envelope staining but COX1 only in cytoplasm. Conclusions In human gastric cancer, there is stronger expression of COX2 than COX1, and different distribution of the two isoforms implies their distinct roles in cell function.

Objective To sum up main syndrome differentiation points of liver depression and spleen asthenia syndrome of chronic hepatitis B(CHB).Methods Logistic regression and discriminant analysis were used for analysis of symptoms,tongue and pulse of liver depression and spleen asthenia syndrome,liver depression syndrome and spleen asthenia syndrome.Results The symptoms selected by logistic regression of liver depression and spleen asthenia syndrome were:abdominal distention or pain,lassitude,poor appetite,loose stool,hypochondriac pain or discomfort and tense pulse;while that selected by discriminant analysis were:abdominal distention or pain,lassitude,loose stool and yellowish fur.Conclusion Main syndrome differentiation points of liver depression and spleen asthenia syndrome of CHB include hypochondriac pain or discomfort,abdominal distention or pain,lassitude,poor appetite and loose stool.Logistic regression and discriminant analysis are feasible for analysis of main diagnosis points of liver depression and spleen asthenia syndrome of CHB.

OBJECTIVE:To observe clinical efficacy and safety of lanthanum carbonate in the treatment of hyperphosphatemia in uremic patients underwent maintenance hemodialysis. METHODS:A total of 142 uremic patients with hyperphosphatemia under-went maintenance hemodialysis were selected from Xinqiao Hospital of Third Military Medical University and Chongqing Banan District Hospital of TCM during Jan. 2012-Dec. 2016. They were given Lanthanum carbonate chewable tablets with initial dose of 250 mg,tid,when serum phosphate level ranged 1.78-2.26 mmol/L or with initial dose of 500 mg,tid,when serum phosphate lev-el was >2.26 mmol/L. Drug dosage was adjusted according the level of serum phosphate during treatment. After 1,2,3 months of treatment,the levels of serum phosphate,serum calcium,albumin,parathyroid hormone(PTH)and alkaline phosphatase were de-tected and the corrected product of serum calcium and calcium phosphorus was calculated. The occurrence of ADR was observed. RESULTS:After 1,2,3 months of treatment,the levels of serum phosphate and the product of serum calcium and calcium phos-phorus were all decreased significantly compared to before trreatment,with statistical significance (P<0.05). After 3 months of treatment,the level of corrected serum calcium was decreased significantly compared to before treatment,with statistical signifi-cance(P<0.05). There was no statistical significance in the levels of plasma albumin,calcium phosphorus or PTH before and af-ter treatment (P>0.05). Total response rate was 93.7%. There were 19 cases of patients with mild adverse reactions. CONCLU-SIONS:Lanthanum carbonate in the treatment of hyperphosphatemia in uremic patients underwent maintenance hemodialysis shows good clinical response rate,can significantly reduce the levels of serum phosphate and the product of serum calcium and calcium phosphorus with good safety.

Objective To investigate the bone marrow supernatant expression of sclerostin in the patients with multiple myeloma (MM) and to clarify its clinical signification .Methods The sclerostin level was quantified by using ELISA ,and the gene expression of sclerostin was determined by RT-PCR .Results The sclerostin level was (0 .54 ± 0 .21)pg/mL in the MM group ,which was sig-nificantly higher than (0 .31 ± 0 .06)pg/mL in the control group (t=5 .67 ,P<0 .01) .The sclerostin level was (0 .65 ± 0 .17) pg/mL in the recurrent and refractory MM group ,which was significantly higher than (0 .47 ± 0 .21) pg/mL in the control group and the newly diagnosed group (t=8 .44 ,3 .27 ,P<0 .01) ,RT-PCR verified that the BMMNC of most patients expressed sclerostin gene .The expression of sclerostin in the MM group was negatively correlated with alkaline phosphatase (ALP)(r= -0 .379 ,P=0 .005) ,and positively related with the correction blood calcium ,bone loss points ,serum β2-micro globulin(β2-GM ) ,proportion of serum M protein and clinical International Staging System (ISS) stages .The median follow-up periods were 29(6-65) months ,the low sclerostin group had the median survival period of 48(6-65) months and the high sclerostin group had the median survival pe-riod of 24(6-52) months ,the difference between them had statistical significance (χ2 = 12 .74 ,P< 0 .01) .Conclusion Its level may reflect the bone destruction ,osteogenesis inhibition degree and myeloma burden ,and reflect the median survival period of MM patients to some extent .

Objective: To understand the effect of Carvedilol on mitochondrial oxidative phosphorylation function during the development of pressure overload induced left ventricular hypertrophy in rats and its mechanism.Methods: Male SD rats were randomized into 6 groups: 5-and 15-week coarctation of the abdominal aorta(H5,H15),5-and 15-week Carvedilol intervention(HR5,HR15) and 5-and 15-week sham operation(S5,S15).Hemodynamics and ventricular remodeling parameters were measured,and the mitochondrial respiratory function was detected by Clark oxygen electrode.Results: Compared with S5,mitochondrial state 3 and 4 respiration and the oxidative phosphorylation rate(OPR) were increased and the respiratory control rate(RCR) decreased significantly in the H5 group.In comparison with S15,state 3 respiration,OPR and RCR were reduced significantly in the H15group.Carvedilol increased the three parameters and restored them to the level of the S15.Conclusion: Mitochondrial respiratory function decreased during the development of pressure overload induced left ventricular hypertrophy in rats.Carvedilol could protect mitochondrial respiratory function and improve myocardial energy metabolism,which might be a mechanism underlying its protective effect on myocardium.

Objective This research is to observe the treatment effects of hip bath and oxygen blowing on severe neonatal diaper rash.Methods 289 neonates with severe diaper rash were randomly divided into three groups.91 cases in the flat tube group,102 cases in the round tube group and 96 cases in the control group.Three groups of neonates were cleaned on the hips and perineuma after poops and then dried with wet tissues and Mupirocin Ointment.The control group was treated with the above-mentioned method.The oxygen blowing groups were treated with hip bath of 1:5000 chameleon solution twice a day.Blowing the hips with oxygen five minutes every time after hip bath.The method of blowing oxygen was that oxygen humidifying containers as normal oxygen aspiration facilities was not filled with water,whose oxygen flowing volume standed at 10 L/min and whose tube blew at the afflicted parts until being dry.The oxygen blower held the oxygen exit and blowed at the afflicted parts in the round tube group and the oxygen blower flattened the oxygen exit and blew at the afflicted parts in the fiat tube group.The treatment effects will be compared among the three groups four days later.Results The cure period of the round tube group was obviously shorter than that of the control group,and the cure period of the fiat tube group was remarkably shorter than that of the round tube group.The total effective rate in the round tube group was obviously higher than that of the control group and the total effective rate in the fiat tube group was obviously higher than that in the round tube group.The difference had a statistical significance.Conclusions The treatment effects for the severe neonatal diaper rash with hip bath and oxygen blowing are remarkable and the oxygen blowing effects with fiat tubes are better than those with round tubes.

Objective To study the expression of the fused proteasome activator REGγ(11S regulator complex gamma subunit) using gene recombination technology and to further study the interaction between REGγ and casein kinase-2 interacting protein-1(CKIP-1)in vitro.Methods Firstly, the full length cDNA fragment of REGγ was amplified through PCR using the plasmid pCMV-Myc-REGγ as template and subcloned into the prokaryotic expression vector pGEX-4T-2 before being transformed into E.coli BL21 cells. The protein expression was induced by isopropyl-β-D-thiogalactoside(IPTG) .Secondly, the protein expression was monitored by SDS-PAGE and Western blotting after ultrasonication. Finally, the GST Pull-down assay was performed to investigate the interaction between REGγ and CKIP-1 in vitro.Results The prokaryotic expression construct pGEX-4T-2-REGγ was generated successfully and confirmed by DNA sequencing. Expression analysis showed that the GST-REGγ protein was easily expressed and isolated mainly in the lysate supernatant after sonication and centrifugation. The GST Pull-down assay revealed the strong mutual interaction between REGγ and CKIP-1 in vitro.Conclusion The proteasome activator REGγ could interact with the negative regulator of osteoblastogenesis CKIP-1 in vitro and the current study has shed light on further investigations of their physiological relevance.