A living cell-based fluorescent reporter for high-throughput screening of anti-tumor drugs / 药物分析学报

Suppression of cellular O-linked β-N-acetylglucosaminylation (O-GlcNAcylation) can repress prolifera-tion and migration of various cancer cells,which opens a new avenue for cancer therapy.Based on the regulation of insulin gene transcription,we designed a cell-based fluorescent reporter capable of sensing cellular O-GlcNAcylation in HEK293T cells.The fluorescent reporter mainly consists of a reporter (green fluorescent protein (GFP)),an internal reference (red fluorescent protein),and an operator (neuronal differentiation 1),which serves as a "sweet switch" to control GFP expression in response to cellular O-GlcNAcylation changes.The fluorescent reporter can efficiently sense reduced levels of cellular O-GlcNAcylation in several cell lines.Using the fluorescent reporter,we screened 120 natural products and obtained one compound,sesamin,which could markedly inhibit protein O-GlcNAcylation in HeLa and human colorectal carcinoma-116 cells and repress their migration in vitro.Altogether,the present study demonstrated the development of a novel strategy for anti-tumor drug screening,as well as for con-ducting gene transcription studies.

Arthroscopy-guided core decompression and bone grafting combined with selective arterial infusion for treatment of early stage avascular necrosis of femoral head / 中国骨伤

<p><b>OBJECTIVE</b>To observe the clinical effects of arthroscopy-guided core decompression and bone grafting combined with selective arterial infusion for early stage avascular necrosis of femoral head.</p><p><b>METHODS</b>From January 2010 to December 2014, 76 patients(76 hips) diagnosed as Ficat II stage avascular necrosis of femoral head were randomly divided into experimental group and control group. In the experimental group, there were 27 males and 8 females aged from 24 to 55 years old with an average of (43.96±6.81) years, treated with arthroscopic-guided core decompression and bone grafting combined with selective arterial infusion. Along the direction of the femoral neck, an 8 mm-diameter tunnel to necrotic areas was drilled, then curettage of necrotic bone was performed under arthroscope, and the iliac bone was grafted. In the control group, there were 29 males and 12 females aged from 26 to 56 years old with an average of (44.62±7.33) years, treated with percutaneous core decompression combined with selective arterial infusion. The preoperative and postoperative Harris scores were recorded and the changes of X-rays were analyzed.</p><p><b>RESULTS</b>All the patients were followed up with an average of 30 months. Postoperative follow-up at 12 months showed that there was significant difference in imaging outcome between two groups(<0.05), the experimental group was better than that of control group. According to Harris hip score system, at the final follow-up, Harris score of the experimental group was 86.72±4.37 on average, 6 cases got excellent results, 24 good, 4 fair and 1 poor. Harris score of the control group was 78.62±5.62 on average, 2 cases got excellent results, 20 good, 15 fair and 4 poor. After Ridit analysis, there was significant difference in the effect between the two groups(<0.05). By pairing sampletest, there was significant difference between the preoperative and postoperative Harris score in the both groups(<0.05). Between the two groups, there was no significant difference in preoperative Harris score(>0.05), but there was significant difference in postoperative Harris score(<0.05).</p><p><b>CONCLUSIONS</b>The two surgical procedures for early femoral head necrosis are effective. Using arthroscopic-guided core decompression method, the necrotic bone can be positioned and scraped more accurately, and can obtain better results.</p>

Observation on the effect of Kangfu Xiaoyan suppository combined with ceftriaxone sodium in treating chronic pelvic inflammatory disease / 中国生化药物杂志

Objective To observe the clinical efficacy of treatment of chronic pelvic inflammatory disease with Kangfu Xiaoyan suppository combined ceftriaxone.Methods84 cases of chronic pelvic inflammatory disease in our hospital from June 2015 to August 2016 were randomly divided into observation group and control group, with 42 cases in each group.The control group were treated with ceftriaxone sodium alone,on the basis of the control group, the observation group was treated with Kangfu Xiaoyan suppository.After treatment, the changes tumor necrosis factor-α (TNF-α), transforming growth factor β1 (TGF-β1), interleukin 8,1β (IL-8, IL-1β) and clinical symptom score in the two groups were recorded and analyzed.Results①After two weeks of treatment, the total effective rate of observation group was 92.86%, significantly higher than that of the control group(76.19%), and the difference was statistically significant (P<0.05).②After treatment,the levels of TNF-α, TGF-β1 in the observation group[(0.80±0.23)μg/L, (12.43±1.98)μg/mL) were better than the control group[(1.81±0.35)μg/L,(19.01±2.24)μg/mL], and the differences were statistically significant (P<0.05);③After treatment,the abdominal pain, vaginal symptom score of observation group[(1.57±0.11),(1.45±0.15) points] were lower than the control group[(2.05±0.17),(2.16±0.20) points], and the differences were statistically significant (P<0.05).ConclusionKangfute Xiaoyan Suppository combined with ceftriaxone sodium in the treatment of chronic pelvic inflammatory disease, curative effect is exact, compared with the single application of ceftriaxone sodium in the treatment without obvious adverse reactions, can effectively improve the symptoms of patients with chronic pelvic inflammatory disease, it is worthy of further research and application.

Application of optical coherence tomography in thyroid surgery / 中华内分泌外科杂志

Objective To investigate the feasibility of using optical coherence tomography (OCT)in thy-roid surgery .Methods Surgical specimen including parathyroid gland , lymph nodes , adipose tissue and thyroid tissue from patients receiving thyroid surgery in Tianjin Medical University Cancer Institute and Hospital were col -lected.OCT images were collected and the results were compared with the routine examination .Results Para-thyroid glands , lymph nodes , adipose tissue , and thyroid tissue can be obtained high-resolution OCT images .Dif-ferent tissues and metastatic lymph node can be clearly distinguished .OCT imaging took about 1 minute and fro-zen examination cost about 25 minutes.Conclusions OCT is a time-saving, non-invasive and cross-sectional imaging technique .It can obtain clear structural images of the internal microstructure of bio-tissues.OCT technol-ogy used in thyroid surgery including parathyroid identification and determination of lymph node metastasis is fea -sible.

A prospective randomized control clinical trial about clopidogrel combined with warfarin versus clopidogrel alone in the prevention of restenosis after femoral-popliteal artery angioplasty / 中华外科杂志

<p><b>OBJECTIVE</b>Using two antithrombotic treatment (clopidogrel vs. clopidogrel combined warfarin) strategies after femoral-popliteal artery angioplasty prospectively, to evaluate which strategy is more effective for the restenosis prevention.</p><p><b>METHODS</b>Totally 50 patients referred for endovascular treatment (including the percutaneous transluminal angioplasty (PTA) and stent implantation) of the superficial femoral artery and popliteal artery from January 2008 to May 2009 were randomly divided into clopidogrel group (group A, 25 cases, 30 limbs) and clopidogrel plus warfarin group (group B, 25 cases, 33 limbs) before operation. Clinical outcomes and restenosis rate of the target lesions were evaluated at 3, 6 and 12 months after operation.</p><p><b>RESULTS</b>Totally 88 patients were screened for participation in the study, 56 patients were included after the follow-up of 12 months. At 3 months, the rates of restenosis were 16.7% in group A and 18.2% in group B (χ² = 0.025, P = 0.874). At 6 months, the accumulated restenosis rates were 36.7% in group A and 36.4% in group B (χ² = 0.001, P = 0.98). At 12 months, the accumulated restenosis rates were 53.3% in group A and 42.4% in group B (χ² = 0.75, P = 0.387). Analysis for the critical limb ischemia sub-group showed that follow-up of 12 months, the accumulated restenosis rate was 8/10 in group A and 6/12 in group B (χ² = 1.023, P = 0.312).</p><p><b>CONCLUSION</b>The clopidogrel alone treatment for PTA or PTA plus stent implantation of femoral popliteal artery has no statistically significant difference in comparison with the clopidogrel combined warfarin treatment in terms of the cumulative vascular restenosis rate at 3, 6, 12 months postoperatively.</p>

Detection on Phenotype of Extended-spectrum ?-Lactamases and Genotype of ?-Lactamases in Klebsiella pneumoniae / 中华医院感染学杂志

OBJECTIVE To investigate the produce of extended-spectrum ?-lactamases(ESBLs) and the presence of genotype of the ?-lactamases-encoding genes in Klebsiella pneumoniae isolated from the 98th Hospital of PLA,Huzhou,Zhejiang Province,China.METHODS Twenty-five strains of K.pneumoniae were isolated from the inpatients between Sep 2005 and Apr 2006.ESBLs were tested by phenotypic confirmatory tests recommended by CLSI.Twenty-one kinds of ?-lactamases genes of blaTEM,blaSHV,blaLEN,blaOKP,blaCTX-M-1 group,blaCTX-M-2 group,blaCTX-M-9 group,blaOXA-1 group,blaOXA-2 group,blaOXA-10 group,blCARB,blaPER,blaVEB,blaGES,blaLAP,blaDHA,blaACT/MIR,blaCMY/MOX,blaFOX,blaCMY/LAT,and blaACC were analyzed by PCR and verified by DNA sequencing.RESULTS In 25 strains of K.pneumoniae,the positive,negative,and "uncertainty" rates of ESBLs were 56.0%,20.0%,and 24.0%,respectively.The positive rate of genes of blaTEM,blaSHV,blaCTX-M-1 group,blaOXA-10 group,blaLAP,and blaDHA were 80.0%,4.0%,4.0%,80.0%,4.0% and 32.0%,respectively.The 15 kinds of rest genes were all tested negative.The total positive rate of 21 kinds of ?-lactamases gene was 92.0%.Among them,the blaLAP-2 gene sequence of the HZ12593 strain has been registered in GenBank(GenBank Accession Number: EU529981).CONCLUSIONS There are higher rate of ESBLs-producing strains in K.pneumoniae isolated from the inpatients,and at least 6 kinds of ?-lactamases gene existed.Both genes of blaTEM and blaOXA-10 group are the most common genotypes.Carring blaDHA Gene may influence the result of phenotypic confirmatory test for ESBLs in K.pneumoniae.

Emergence of 16S rRNA methylase gene rmtB in Klebsiella pneumoniae isolates from the inpatients in Zhejiang province, China / 中华流行病学杂志

Objective To investigate the presence and genetic background of 16S rRNA methylase gene and Aminoglycoside modifying enzymes(AMEs) genes in Klebsiella pneumoniae isolated from the People's Liberation Army 98th HospitaI,Huzhou district,Zhejiang province,China.Methods 25 strains of Klebsiella pneumoniae were isolated from the inpatienta between September,2005 and April,2006.6 kinds of 16S rRNA methylase gene (including armA,rmtA,rmtB,rmtC,rmtD and npmA ),6 kinds of AMEs genes [ including aac (3)-Ⅰ,sac (3)-Ⅱ,sac (6')-Ⅰ,aac (6')-Ⅱ,ant (3")-Ⅰand ant(2")-Ⅰ],intI1,intI2,intI3,mercuric reductase gene merA (merA gene were the collective genetic markers of transposona of Tn21 and Tn501 ) and tnpA ( tnpA gene were the collective genetic markers of transposons of Tn1,Tn2,Tn3 and Tn1000) were analyzed by PCR and verificated by DNA sequencing.Results In 25 strains of Klebsiella pneumoniae,the positive rate of genes of rmtB,sac (3)-Ⅱ,sac (6')-Ⅰ,ant(3")-Ⅰ and intI1 were 60.0%(15/25),4.0%(1/25),48.0%(12/25),60.0%(15/25) and 96.0%(24/25),respectively.The rest 12 kinds of genes were all tested negative.The total positive rate of 6 kinds of AMEs gene was 84.0%(21/25).Conclusion There were very high positive rate on both genes of rmtB and AMEs genotypes in Klebsiella pneumoniae isolated from inpatients,and this was the first report of the emergence of 16S rRNA methylase gene rmtB in Klebsiella pneumoniae identified in Zhejiang province,China.

Analysis on 16S rRNA methylase genes and aminoglycoside modifying enzymes genes in Enterobacter cloacae in China / 中华流行病学杂志

Objective To investigate the 16S rRNA methylase genes and Aminoglycoside modifying enzymes(AMEs)genes in Enterobacter cloacae isolated from the People's Liberation Army 98th Hospital,Huzhou district,Zhejiang province,China.Methods 40 strains of Enterobacter cloacae were isolated from the inpatients between September,2003 and November,2004.5 kinds of 16S rRNA methylase gene (including armA,rmtA,rmtB,rmtC and rmtD)and 9 kinds of AMEs gene[including aac(3)-Ⅰ,aac(3)-Ⅱ,aac(3)-Ⅲ,aac(3)-Ⅳ,aac(6')-Ⅰ b,aac(6')-Ⅱ,ant(3'')-Ⅰ,ant(2'')-Ⅰ and aph(3')-Ⅵ]were analyzed by PCR and verificated by DNA sequencing.Results In 40 strains of Enterobacter cloacae,the positive rates of genes of rmtB,aac(3)-Ⅱ,aac(6')-Ⅰ b,ant(3'')-Ⅰ,ant(2'')-Ⅰ and aph(3')-Ⅵ were 12.5%(5/40),27.5%(11/40),72.5%(29/40),32.5%(13/40),5.0%(2/40)and 5.0%(2/40),respectively.8 kinds of the rest of genes were all tested negative.The total positive rate of AMEs gene was 85.0%(34/40).Among 29 strains of Enterobacter cloacae that the aac(6')-Ⅰ b gene was positive,through PCR and verification by DNA sequencing,7 strains(24.1%)were confirmed to take the aac(6')-Ⅰ b-cr(the GenBank register number:EF375620,EU159121)alone,18 strains(62.1%)were confirmed to take the aac(6')-Ⅰ b-Suzhou(EU085533)alone,3 strains(10.3%)were confirmed to take both aac (6')-Ⅰ b-Suzhou and aac(6')-Ⅰ b-cr while only 1(3.4%)was aac(6')-Ⅰ b(the classical type).Conclusion There was lower positive rate of 16S rRNA methylase gene but very high AMEs genotypes in Enterobacter cloacae isolated from inpatients and the finding of rmtB gene was reported for the first time in the world.At least 5 kinds of AMEs gene existed in Enterobacter cloacae were isolated and they were the new host of both gene of aac(6')-Ⅰ b-cr and aac(6')-Ⅰ b-Suzhou,with aac(6')-Ⅰ b-Suzhou gene was the predominance subtype in aac(6')-Ⅰ b.

?-Lactamase Genes in Four Kinds of Nonfermenting Gram-negative Bacilli / 中华医院感染学杂志

OBJECTIVE To investigate the ?-lactamase genes in 4 kinds of nonfermenting Gram-negative bacilli isolated from the 98th Hospital of PLA. METHODS Sixty strains of Acinetobacter baumannii,30 strains of Pseudomonas aeruginosa,19 strains of Stenotrophomonas maltophilia,and 15 strains of Flavobacterium were isolated from hospitalized patients.Nine kinds of ?-lactamases genes of TEM,SHV,OXA,CTX-M,PER,VEB,IMP,VIM and GES were analyzed by PCR and verified by DNA sequencing. RESULTS In A.baumannii and(P.aeruginosa),the positive rates of gene of TEM were 100.0% and 66.7%,respectively.SHV gene was positive in 18 of 60 strains of A.baumannii tested,17 of which were SHV-12 subtype ESBLs.The other was a new SHV type ?-lactamase nominated SHV-48.OXA gene was positive in 1 of 30 strains of P.aeruginosa tested,it was an OXA-10 subtype ESBLs.But the rest of genes were all negative. CONCLUSIONS There exist 4 kinds of(?-lactamase) genes at least in nonfermenting Gram-negative bacilli including TEM-1,SHV-12,SHV-48,and(OXA-10.)

bla_(LAP-2),A Novel Subtype of ?-Lactamases Gene in Enterobacter cloacae in China / 中华医院感染学杂志

OBJECTIVE To investigate the resistance genes in HZB9055 strain of Enterobacter cloacae isolated from the PLA 98th Hospital,Huzhou District,Zhejiang Province,China.METHODS HZB9055 Strain of E.cloacae was isolated from the inpatient in May,2004,41 kinds(or groups) of genes were analyzed by PCR and verified by DNA sequencing.RESULTS In HZB9055 strain of E.cloacae,3 kinds of genes were positive including blaLAP,blaMIR and qnrS.The 38 kinds of rest genes were all tested negative.The blaLAP gene sequence including 858 nucleotides,which had the amino acid mutation in position 193 compared with LAP-1 type narrow-spectrum ?-lactamase(GenBank accession number:EF026092),and was nominated LAP-2(GenBank accession number:EU159120).CONCLUSIONS At least 3 kinds of resistance genes exist in HZB9055 strain of E.cloacae,and blaLAP-2 is a novel subtype of ?-lactamases gene.

Antimicrobial-resistant Genes and Consanguinity in Multi-resistant Pseudomonas aeruginosa / 中华医院感染学杂志

OBJECTIVE To investigate the antimicrobial-resistant genes and consanguinity in multi-resistant Pseudomonas aeruginosa(MRPA) isolated from the 98th Hospital of PLA,Huzhou,Zhejiang Province,China.METHODS Thirty strains of MRPA were isolated from hospitalized patients between Sep 2003 and Oct 2004.Twenty four kinds of genes of blaTEM,blaSHV,blaOXA-10 group,blaPER,blaVEB,blaIMP,blaVIM,blaGES,blaCARB,blaDHA,blaMIR,aac(3)-Ⅰ,aac(3)-Ⅱ,aac(3)-Ⅲ,aac(3)-Ⅳ,aac(6′)-Ⅰ,aac(6′)-Ⅱ,ant(3″)-Ⅰ,ant(2″)-Ⅰ,aph(3′)-Ⅵ,oprD,qacE△1-sul1,catB,and cml1 were analyzed by PCR and verified by DNA sequencing.Resistant-genes cluster analysis was performed by Average.RESULTS In 30 strains of MRPA the positive rate of genes of blaTEM,blaOXA-10 group,blaCARB,aac(3)-Ⅱ,aac(6′)-Ⅰ,aac(6′)-Ⅱ,ant(3″)-Ⅰ,ant(2″)-Ⅰ,qacE△1-sul1,and cml1 were 66.7%,3.3%,3.3%,76.7%,3.3%,33.3%,53.3%,26.7%,83.3%,and 3.3%,respectively,and the deficiency rate of oprD gene was 90.0%.The gene of blaOXA-10 group was sequenced and determined to be blaOXA-10 subtype ESBL gene.But the rest of genes were all negative.According to the cluster analysis of resistant-gene,30 strains of MRPA isolated could be classified into four subgroups,which were caused by the infection in hospital.CONCLUSIONS At least 10 kinds of antimicrobial-resistant genes exist in MRPA isolates,and the deficiency rate of oprD gene is very high.MRPA can induce clone transmitted hospital infection and it has fulminant prevalence.