Objective To characterize the resistance mechanisms of a clinical Shigella sonnei strain harboring blaCTX-M-55 .Methods A double-disk synergy test was conducted to detect ESBL.Antibiotic resistance genes were determined by PCR followed by amplicon sequencing.Conjugation experiments were performed to verify the transferability of the plasmids carrying ESBL genes.The minimum inhibitory concentration values were tested using VITEK 2.The transposition unit was confirmed by DNA sequencer,and the transcriptional start site was identified using primer extension assay.Results Strain #1083 produced CTX-M-55,which was encoded by plasmid p1083-CTXM that could be transferred into E.coli through conjugation experiments to confer corresponding antibiotic resistance to the transconjugant #1083-EC600.The transposition unit mediating the transfer of blaCTX-M-55 was ISEcp1-blaCTX-M-55 -Δorf477.ISEcp1 offered strong promoter regions for the resistance genes,facilitating their expressions.Besides,the expressions were constant,not induced by antibiotics.Conclusion BlaCTX-M-55 on plasmids is the major resistance genes for strain #1083.Their expressions and spread are mediated by the insertion sequence ISEcp1.

BACKGROUND:MicroRNA plays an important role in the process of growth and aging of living body. To know the role of let-7d in inducing bone marrow mesenchymal stem celldifferentiation into neurons can promote the stem celltransplantation. OBJECTIVE:To investigate the role of let-7d in inducing bone marrow mesenchymal stem celldifferentiation into neurons. METHODS:(1) The lentiviral vector of let-7d was constructed and transfected into rat bone marrow mesenchymal stem cells. The cells were divided into non-transfected group, negative control group (transfected with FU-RNAi-NC-LV), transfected enhancement group (transfected with let-7d-LV), transfected inhibition group ( transfected with let-7d-inhibition-LV). (2) Rat bone marrow mesenchymal stem cells were treated with fasudil as an inducer for triggering the cells to differentiate into neurons. The expression of neuron-specific markers, neuron-specific enolase and microtubule-associated protein 2, were measured by immunocytochemical method. The mRNA expression of microtubule-associated protein 2 was detected by RT-PCR. The viability of bone marrow mesenchymal stem cells was determined by MTT method. RESULTS AND CONCLUSION:Under inverted fluorescence microscope, the cells were successful y transfected with let-7d. Fasudil induced bone marrow mesenchymal stem cells to differentiate into neurons. The transfection efficiency and expression levels of neuron-specific enolase and microtubule-associated protein 2 in transfected enhancement group were higher than those in the negative control group (P<0.05);while in the inhibition group, they were lower than those in the negative control group (P<0.05). These findings indicate that let-7d can promote the differentiation of bone marrow mesenchymal stem cells into neurons induced by fasudil, and by control ing the expression of let-7d we can influence the differentiation efficiency from bone marrow mesenchymal stem cells to neurons.

OBJECTIVETo evaluate the intermediate and long-term follow-up effect of posterior dynamic lumbar stabilization in lumbar degenerative disease.

METHODSThe clinical outcomes of 96 patients (male 51, female 45, age from 21 to 68 years, mean 41.5 years) whose follow-up time were more than 2 years with lumbar degenerative disease treated by posterior decompression with Wallis posterior dynamic lumbar stabilization implant or combined with posterior lumbar fusion from August 2007 to January 2010 were retrospectively studied, and assessed with visual analogue scale (VAS) and spinal operative standard of Chinese Medical Association. The early and long-term follow-up effect and complications associated with Wallis posterior dynamic lumbar stabilization were recorded. The height of intervertebral space at the treated level in lateral plain film were measured at preoperatively, 3 month postoperatively and last follow-up, respectively. The finds of MRI obtained at over 6 month postoperative were recorded.

RESULTSThe operative procedure of Wallis posterior dynamic lumbar stabilization implant was easy and less invasive. The VAS scores were 78 ± 24, 28 ± 16 and 14 ± 12 preoperatively, 3 month postoperatively and last follow-up, respectively. The good or excellent result was 91.7% at the last follow-up. No complication related with Wallis posterior dynamic lumbar stabilization was found. The rate of patient's satisfaction with the Wallis implant operation was 95.8%. The disc height at the treated level in lateral plain film were (8.2 ± 3.7), (10.4 ± 2.6) and (10.1 ± 1.9) mm at preoperatively, 3 month postoperatively and last follow-up, respectively. There is no further degenerative change found in MRI obtained at over 6 month postoperative. MRI 1 year after Wallis procedure showed rehydration of the formerly black disc at the treated level.

CONCLUSIONSIt is easy and safe to use Wallis posterior dynamic lumbar stabilization in treatment of degenerative lumbar disease, and the effect of the intermediate and long-term follow-up more than 2 years is good. The Wallis system provides an alternative for treatment of lumbar degenerative disease.

Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Internal Fixators ; Intervertebral Disc Degeneration ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Spinal Fusion ; instrumentation ; methods ; Treatment Outcome ; Young Adult

OBJECTIVETo observe the relation of androgen levels and atherosclerosis (AS) in elderly males.

METHODSBoth carotid arteries and arteries of lower extremity were examined with Doppler ultrasonography. Those with arteriosclerosis and much atheromatous plaque were designated as case group, and those with normal results formed control group. Total testosterone (TT), free testosterone (FT) and estradiol (E2) were measure by radioimmunoassay, HDL-C, LDL-C, TC and TG were assayed by colorimetry, vascular endothelium growth factor (VEGF), tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), intercellular adhesion molecule (sICAM-1) were determined by ELISA.

RESULTFT was significantly lower in case group than in control group (P<0.01), no differences were found in TT, E2. HDL-C in control group was higher than that in case group (P<0.01), TC and TG were higher in case group than those in control group (P<0.05). HDL-C was correlated positively and LDL-C was negatively with FT level, while both TC and TG in case group had negative relation with FT. VEGF was higher in case group (P<0.05), and it had negative relation with FT in both groups. TNF-alpha and IL-6 were significantly higher in case group (P<0.05), and they had negative relation with FT. sICAM-1 was significantly lower in control group than it in case group (P<0.01).

CONCLUSIONThe normal androgen levels, especially FT, have beneficial effect in AS development in elderly males. Low FT level may be an independent risk factor in AS development.

Aged ; Androgens ; blood ; Atherosclerosis ; blood ; diagnostic imaging ; Carotid Arteries ; diagnostic imaging ; Humans ; Male ; Middle Aged ; Risk Factors ; Testosterone ; blood ; Ultrasonography, Doppler

OBJECTIVETo observe the expression of Resistin mRNA in peripheral blood mononuclear cells and its gene polymorphism in coding region in a small range population in Zhejiang Province of China.

METHODSEighty-three cases of type 2 diabetes mellitus and 53 healthy people were included. The expression of Resistin mRNA in peripheral blood mononuclear cells was detected by RT-PCR and semi-quantitative PCR assay. The sequencing work was done in Resistin cDNA and gene polymorphism was analyzed.

RESULTSAt the same condition, in 83 diabetes patients, Resistin mRNA was detected in 23 cases (11 males and 12 females). There was no Resistin mRNA expression in 53 healthy people. The ratio of PCR products between Resistin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was from 0.564 to 1.238, averaging 0.804+/-0.436. The sequence of Resistin cDNA is almost identical with each other and with that in GenBank with no single nucleotide polymorphism being found.

CONCLUSIONResistin mRNA is expressed in human peripheral blood mononuclear cells in some type 2 diabetes mellitus, but its expression is at a low level. Among the experiment population we did not find polymorphism phenomenon in Resistin coding region. The different individual's Resistin coding region is highly coincident.

China ; epidemiology ; DNA Mutational Analysis ; Female ; Gene Expression ; genetics ; Gene Frequency ; Genetics, Population ; Humans ; Leukocytes, Mononuclear ; Male ; Middle Aged ; Polymorphism, Genetic ; Polymorphism, Single Nucleotide ; genetics ; Resistin ; genetics

OBJECTIVETo evaluate clinical outcomes after laparoscopic total mesorectal excision (TME) combined with intersphincteric resection (ISR) for ultra-low rectal tumors.

METHODSClinical data of 36 patients with ultra-low rectal tumor undergoing laparoscopic TME combined with ISR were analyzed retrospectively.

RESULTSThe median distance from the inferior margin of the tumor to the anal verge was 3.4 (2.0-5.0) cm. There were 33 cases of well/moderately differentiated adenocarcinoma and 3 rectal malignant villous adenoma. There were 16 patients with stage I disease, 15 with stage II A, 3 with stage III A, and 1 with III B. Postoperatively, one patient developed stenosis at the end ileostomy and 3 anastomotic leakage. After a median follow-up of 16(4-49) months, one patient developed local recurrence at the anastomosis and one case died of liver metastasis. In the 19 patients who had a minimum follow-up of one year, the bowel movements frequency ranged from 1-4 times per day, and these patients were able to withhold defecation for more than 5 minutes.

CONCLUSIONSLaparoscopic TME combined with ISR can achieve oncologic clearance, sphincter preservation, and minimal invasiveness for ultra-lower rectal cancer. However, patients selection should be cautious.

Adult ; Aged ; Aged, 80 and over ; Anal Canal ; surgery ; Female ; Follow-Up Studies ; Humans ; Laparoscopy ; Male ; Mesentery ; surgery ; Middle Aged ; Rectal Neoplasms ; surgery ; Rectum ; surgery ; Retrospective Studies ; Treatment Outcome

Objective:To detect gene mutations in two patients with xeroderma pigmentosum (XP) .Methods:Clinical data were collected from two patients with XP. DNA was extracted from peripheral blood of the patients and their parents, and high-throughput whole-exome sequencing was performed in the patient to identify pathogenic gene mutations. Then, Sanger sequencing was performed to bidirectionally verify the mutations in the patients and their parents, with a focus on mutations in the candidate genes XPA, ERCC3, XPC, ERCC2, DDB2, ERCC4, ERCC5 and POLH.Results:Case 1, a 3-year-old male patient, presented with brown spots and hypopigmented macules on the face, ears, neck and back of both hands for 2 years, as well as gait instability for 1 year. The skin lesions were mainly distributed in sun-exposed areas and worsened in summer. Case 2, a male patient aged 1 year and 5 months, presented with scattered brown spots and a few hypopigmented macules on the face for 1 year. Genetic testing showed compound heterozygous mutations in the ERCC2 gene of case 1, including c.1805G>A (p.Gly602Asp) mutation inherited from his father and c.586C>T (p.Arg196Ter) mutation inherited from the his mother, so case 1 was diagnosed with XP group D (XPD) . Compound heterozygous mutations were identified in the ERCC5 gene of case 2, including c.2533+2T>C mutation inherited from his father and c.2453C>T (p.Ala818Val) mutation inherited from his mother, so case 2 was diagnosed with XP group G (XPG) . The c.586C>T (p.Arg196Ter) and c.2533+2T>C mutations had not been reported before. Sun protection was suggested. After a follow-up of 2 years, skin lesions increased in the 2 patients, but no malignant tumors occurred.Conclusions:The compound heterozygous mutations in the ERCC2 gene can lead to XPD, mainly manifesting as cutaneous and neurological symptoms, while the compound heterozygous mutations in the ERCC5 gene can lead to XPG, manifesting as a mild phenotype. Early clinical features combined with genetic testing are helpful for accurate diagnosis and classification of XP.

Objective To analyze the outcome indexes selected in the randomized controlled trials(RCTs)on traditional Chinese medicine(TCM)for skin rash caused by EGFR-TKI.Methods Seven databases were researched and the literature was screened according to the inclusion and exclusion criteria,then summarize and categorize outcome indexes to calculate frequency of outcomes and analyze.Results 46 out of 2241 papers were included.The outcome indexes of 40 RCTs were mainly divided into seven categories,that is physical symptoms/signs(37.97%),quality of life(17.65%),safety indicators(13.37%),traditional Chinese medicine symptoms/syndromes(16.58%),long-term prognosis(1.60%),blood biochemical index(9.09%)and others(3.74%).Problems as follow:First,there is a high risk of clinical trial bias in the treatment of EGFR-TKI-associated rash with TCM.Besides,the measurement time points was largely different with the severity of rash not considered.What's more,oversimplify the reports of compound outcome indicators and safety evaluation,lack of treatment time window and full process observation.Last,grading criteria for rash were inconsistent and the function of quality of life assessment tool was relatively simple.Conclusion At present,the use of outcome indicators has not been standardized in the RCTs for skin rash related EGFR-TKI by TCM,and relevant construction work should be carried out in the future to build a core index set with the characteristics of TCM treatment.

OBJECTIVEUsing nested polymerase chain reaction (PCR) to perform preimplantation gender diagnosis.

METHODSOne (or two) lymphocyte and blastomere (n=50/group) were collected and prepared under the following conditions: (1) water only (H(2)O); (2) freeze-thaw liquid nitrogen, then boiling; (3) potassium hydroxide/dithiotheriol, heated to 65 degree centigrade, followed by acid neutralization (KOH). Cells were analyzed by PCR using nested primers amplification with amelogenin gene.

RESULTSThe amplification rate and allele dropout (ADO) rate for male lymphocytes by the three methods were 83%, 94%, 95% and 24%, 12%, 4%, respectively. Using two cells per reaction did not increase the amplification rate for the KOH method.

CONCLUSIONThe KOH method for DNA preparation is superior to the other methods evaluated. Dual blastomere biopsy and independent blastomere analysis may improve preimplantation diagnostic reliability.

Amelogenin ; Blastocyst ; cytology ; metabolism ; Blastomeres ; cytology ; metabolism ; DNA ; genetics ; Dental Enamel Proteins ; genetics ; Female ; Genotype ; Humans ; Lymphocytes ; cytology ; metabolism ; Male ; Polymerase Chain Reaction ; methods ; Sex Determination Analysis ; methods

OBJECTIVETo evaluate the clinical value of high frequency ultrasound in diagnosing peripheral nerve diseases (PNDs).

METHODSFrom January 2003 to December 2006, 64 cases of PNDs were analyzed retrospectively. The ultrasound diagnosis was compared with the operative and pathological diagnosis.

RESULTSBased on the operative and histopathological results, in 38 patients with trauma or entrapment, 38 among 45 traumatic nerves were rightly diagnosed by ultrasound. The coincidence rate was 84.4%. In 26 patients with original peripheral nerve tumors (PNTs), including 20 neurilemmomas, 4 neurofibromas and 2 malignant neurilemmomas, 16 cases were diagnosed by ultrasound with a coincidence rate of 61.5%. The coincidence rates in limbs and trunk were 86.7% (13/15) and 27.3% (3/11) respectively.

CONCLUSIONSThe study suggests that high frequency ultrasound can locate peripheral nerve trauma precisely, assess the impair degree correctly and provide useful information for clinic diagnosis. The high frequency ultrasound brings better diagnosis outcome in limbs nerve tumors than in trunk.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Male ; Middle Aged ; Peripheral Nerve Injuries ; Peripheral Nervous System Diseases ; diagnostic imaging ; Retrospective Studies ; Trauma, Nervous System ; diagnostic imaging ; Ultrasonography