Objective To evaluate the capability of multilocus variable-number tandem-repeat ( VNTR) analysis ( MLVA) and pulsed-field gel electrophoresis ( PFGE) for genotyping Salmonella enterica serovar Typhi (S.Typhi) isolates.Methods Five polymorphic VNTRs (SAL02,SAL11,SAL16,SAL20, and TR4699 ) that were observed in S.Typhi strains from previous studies were selected to establish MLVA . Twenty-one epidemiologically unrelated S.Typhi strains that were isolated from Shenzhen ,China from 2002 to 2007 were genotyped by the established MLVA , and the results were compared with those by PFGE . Results The Simpson′s index of diversity ( D value ) for all five different VNTRs ranged from 0.838 to 0.943 .A total of 19 MLVA profiles and 19 PFGE profiles were found , respectively .The D value for both MLVA and PFGE were 0.99 and the test results from two analyses were identical .Conclusion The five polymorphic VNTRs analysis could be used as an alternative typing scheme for epidemiologic investigation of S.Typhi infection .

Objective To investigate the prevalence and molecular characteristics of the extended -spectrum β-lactamase ( ESBL) and AmpC enzyme-producing Proteus mirabilis ( P.mirabilis) strains isola-ted in Shenzhen People′s Hospital.Methods The production of ESBLs and AmpC enzymes by P.mirabilis isolates were detected by a screening and confirmatory test for ESBLs and AmpC disk test , respectively .The PCR assays followed by DNA sequencing of the products were employed to analyze the multiple genes inclu -ding the ESBLs genes, AmpC genes, insertion sequences (ISs) upstream of the ESBLs or AmpC genes, plasmid -mediated quinolone resistance ( PMQR ) determinants , quinolone resistance-determining region (QRDR) genes , the integrase genes, and class1 integron cassette.The epidemiological analysis of the iso-lates was performed by pulsed field gel electrophoresis .Results There were 130 P.mirabilis clinical iso-lates collected from Shenzhen People′s Hospital in China during the year 2004 to 2010.Among them, 13 isolates (10%) produced ESBLs, that accounted for 0%-9.1%in the year 2004-2009 and up to 29.4%in 2010, and 3 isolates (2.3%) produced AmpC enzymes.The predominant genotype of ESBLs -producing isolateswas b al CTX-M-14(n=7), followed by blaCTX-M-65(n=3), blaCTX-M-55(n=1), blaCTX-M-24(n=1) and blaPER-1 (n =1).The clinical isolate of PER-1-producing P.mirabilis was reported for the first time in China.Twoisolates carried an AmpC β-lactamase gene of blaCMY-2 and one isolate carried an unidentified AmpC gene .ISEcp1 located upstream of blaCTX-M and blaCMY-2 were detected in 91.7% (11/12) of CTX-M-producing isolatesand one CMY-2-producing isolate, respectively.ISPa12 was present upstream of blaPER-1 in one studiedisolate.Approximately 66.7% (10/15) of ESBL and /or AmpC-producing isolates harbored PMQR genes including2 carrying qnrD, 5 carrying aac-Ib-cr and 3 carrying both qnrD and aac-Ib-cr.Twelve ESBL and /orAmpC-producers with high level of resistance to ciprofloxacin carried the similar mutation profiles of S 83I inGyrA, S80I or S80R in ParC and among them, six strains showed E466D mutation in GyrB.Approximately86.7% (13/15) of ESBL and/or AmpC-producing isolates carried class 1 integron.Fourteen PFGE typeswere observed among 15 ESBL and/or AmpC-producers.Conclusion The prevalence of CTX-M β-lactamasesin P.mirabilis isolates contributed to the increased resistance to extended -spectrum cephalosporins.The qnrD and/or aac-Ib-cr genes were detected among the most of ESBL and /or AmpC-producing P.mirabilis clinical isolates.

The aim of the present study was to investigate the changes in single-channel currents of large conductance calcium-activated potassium channels (BK(Ca) channels) in cerebral vascular smooth muscle cells (VSMCs) of rats after 1-week simulated microgravity. Sprague-Dawley rats were subjected to tail-suspension (SUS) to simulate cardiovascular deconditioning due to microgravity. Cytosolic calcium ([Ca(2+)](i)) was examined by laser-scanning confocal microscopy with calcium-sensitive-dye Fluo-3/AM as fluorescent probe. Single-channel currents of BK(Ca) channels were measured with cell-attached membrane patches bathed in symmetrical high potassium solution. The [Ca(2+)](i)i level was significantly higher in cerebrovascular myocytes of SUS than that of control (CON) rats. The probability of open (Po) and the mean open time (To) of BK(Ca) channels in cerebral VSMCs significantly increased in SUS as compared with CON. However, there were no significant differences in the unitary conductance and mean close time (Tc) between the two groups. The results obtained suggest that both the elevated [Ca(2+)](i) and enhanced single-channel activities of BK(Ca) channels in cerebral VSMCs might be among the electrophysiological mechanisms that mediate the increased vasoreactivity and hypertrophic change in cerebral arteries during adaptation to simulated microgravity in rats.

OBJECTIVETo investigate the injury in the corneal nerve and cornea of rats exposed to n-hexane.

METHODSThirty-two SD male rats were randomly divided into one control group and four n-hexane groups. The four n-hexane groups inhaled 35.2 g/m(3) n-hexane statically for 1, 3, 7 and 14 d respectively, while the rats in the control group inhaled air. The corneal nerve damage was investigated with golden staining and transmission electron microscope. Histopathological and ultrastructure changes of cornea were analyzed also.

RESULTSThe concentration of n-hexane in blood of rats in different experimental groups was (242.91 +/- 59.68), (668.77 +/- 221.74), (1021.21 +/- 545.71) and (1140.42 +/- 468.44) microg/L, increased gradiently with time exposed to n-hexane. In the rats exposed to n-hexane for 7 and 14 d, there appeared fewer corneal nerve bundles and lower density of nerve fiber at the center of cornea, under electron microscope, the lamellar sheath of nerve fiber in the corneal epitheliums appeared intermittent, the neuroplasm of endings was partly lysed and became vacuolar, the microfilament and racuole of neuraxon decreased. In the group exposed to n-hexane for 14 d, the microvillus of cornea epithelium were decreased. In some basal cells there appeared pyknotic nucleus and vacuole, mitochondria were swollen or disappeared.

CONCLUSIONThe structure of corneal nerve and cornea is damaged in the rats exposed to n-hexane, thus leading to dysfunction of cornea.

Animals ; Cornea ; Nerve Tissue ; Rats

The present study was designed to investigate the effects of various extracellular matrix (ECM) proteins on the biological characteristics of late endothelial progenitor cells (EPCs). Density gradient centrifugation-isolated rat bone marrow mononuclear cells were cultured in complete M199 medium, which contained 15% fetal calf serum, 10 μg/L vascular endothelial growth factor (VEGF) and 5 μg/L basic fibroblast growth factor (bFGF). EPCs were plated on substrates containing fibronectin (Fn), laminin (Ln) or rat tail tendon collagen (Col), and the corresponding cells were defined as Fn, Ln and Col groups. The 3rd generation EPCs, namely late EPCs, were harvested. The proliferation, adhesion, migration and the ability of forming tubes were assayed using CCK-8, adhesion test, wound healing assay and Matrigel, respectively. The mRNA expressions of endothelial cell differentiation markers, vWF and CD31, were analyzed by real time RT-PCR. The apoptosis was assayed by flow cytometry (FCM). The results showed that cell proliferation ability of Fn and Col groups were higher than that of Ln group; Fn group showed increased adhesion compared to Col and Ln groups (P < 0.01); The migration ability of Fn and Col groups were higher than that of Ln group. Moreover, Fn group showed increased tube formation abilities compared to Col and Ln groups (P < 0.05). Although 24-hour free-serum-induced apoptosis in Ln group was the highest, there was no difference of auto-apoptosis among the three groups. Furthermore, the mRNA expressions of vWF and CD31 exhibited no difference among the three groups. These results suggest the ECM affects the biological functions of late EPCs, which would have a high probability of providing new directions that lead to the development of artificial heart and blood vessels.
Animals ; Cell Proliferation ; Cells, Cultured ; Collagen ; chemistry ; Endothelial Progenitor Cells ; cytology ; Extracellular Matrix ; physiology ; Extracellular Matrix Proteins ; chemistry ; Fibroblast Growth Factor 2 ; chemistry ; Fibronectins ; chemistry ; Rats ; Vascular Endothelial Growth Factor A ; chemistry

OBJECTIVETo analyze the expression of deleted in liver cancer 1 (DLC1) and phosphorelated focal adhesion kinase (p-FAK) in breast cancer tissue to further understand the molecular mechanisms of the carcinogenesis and metastasis of breast cancer.

METHODSImmunohistochemistry was employed to determine the protein level of DLC1 and p-FAK in 61 breast cancer, 30 benign breast disease and the adjacent normal breast tissues.

RESULTSThe positivity rates of DLC1 differed significantly between breast cancer, benign and normal tissues (34.43%, 80.00% and 76.67%, respectively, P<0.001). The positivity rates of p-FAK in the 3 tissues were 77.05%, 33.33% and 26.67%, also showing significant differences (P<0.001). The aberrant expression of DLC1 showed an inverse correlation to p-FAK (κ=-0.4591). Both DLC1 and p-FAK were closely correlated to the carcinogenesis, clinical stage, PR and lymphatic metastasis of breast cancer (P<0.05), but not to the patients age, pathological subtype, familial history, ER or CerbB-2 (P>0.05).

CONCLUSIONThe abnormal expression of DLC1 and p-FAK might participate in the carcinogenesis, progression, and metastasis of breast cancer. The role of DLC1 and p-FAK might be related to the regulation of progestone. DLC1 and p-FAK may serve as candidate markers for early diagnosis, prognostic evaluation and target treatment of breast cancer.

Adult ; Breast Neoplasms ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Female ; Focal Adhesion Kinase 1 ; metabolism ; GTPase-Activating Proteins ; metabolism ; Humans ; Lymphatic Metastasis ; Middle Aged ; Phosphorylation ; Prognosis ; Receptors, Progesterone ; metabolism ; Tumor Suppressor Proteins ; metabolism

OBJECTIVETo study the efficacy of metformin intervention on insulin resistance during catch-up growth in mice with fetal growth restriction (FGR).

METHODSMouse models of FGR were established by low protein diet feeding of the pregnant mice. Both the newborn female mice with FGR and normal control (NC) mice were randomized for feeding with a standard diet (SF) or a high-fat diet (HF) after weaning and treatment with gavage of either metformin or normal saline. The mice were examined for vaginal opening time and the estrous cycle at the age of 8 weeks. At the age of 12 weeks, 6 mice in anestrus from each group were fasted for 12 h for measurement of body weight, height, poundera index (PI), fasting blood glucose (FBG), fasting insulin (Fins), follicle stimulating hormone (FSH) and anti-Mullerian hormone (AMH), and the HOMA-IR was calculated. The reproductive capacity of female mice was assessed by mixing them with male mice at the ratio of 2:1. The 3 × 2 factorial analysis was conducted to determine the interactions between FGR, high-fat feeding and metformin.

RESULTSFactorial analysis showed that FGR and high-fat feeding had significant effects on the PI index, Fins, HOMA-IR, vaginal opening time, and AMH (P<0.05). Metformin significantly affected the factors related to high-fat feeding including weight, PI, FPG, Fins, HOMA-IR and estrous cycle (P<0.05) and the factors related to FGR with the exception of height and FSH (P<0.05). FGR significantly affected the factors tested except for body weight (P<0.05); high-fat feeding affected all the factors but the FSH (P<0.05); metformin affected all the factors but the height and FSH (P<0.05). In the female mice treated with saline, the pregnancy rates differed significantly between FGR mice with high-fat feeding and control mice with standard feeding, and between FGR mice with standard feeding and high-fat feeding (P<0.05).

CONCLUSIONFGR mice can present with delayed puberty with rare ovulation and adulthood insulin resistance, and high-fat feeding after birth can promote the catch-up growth of FGR mice. Metformin intervention is effective for improving insulin resistance and reproductive-endocrine disorders in FGR mice during catch-up growth.

We propose a method using total variation (TV) regularization in deconvolution for partial volume correction in PET imaging. In the degraded image model, we used TV regularization procedure in Van Cittert (VC) and Richardson-Lucy (RL) deconvolution algorithms. These methods were tested in simulated NCAT images and images of NEMA NU4-2008 IQ phantom and tumor-bearing mouse scanned by Simens Invoen microPET. The simulated experiment and tumor-bearing mouse experiment showed that the algorithms using TV regularization provided superior qualitative and quantitative appearance compared with traditional VC and RL algorithms. When the mean intensity of the tumor increased by (10±1.8)%, the SD increase percentage was decreased from 49.98% to 14.26% and from 42.76% to 4.70%, suggesting the efficiency of the proposed algorithms for reducing PVEs in PET.
Algorithms ; Animals ; Image Processing, Computer-Assisted ; Imaging, Three-Dimensional ; Mice ; Phantoms, Imaging ; Positron-Emission Tomography

OBJECTIVETo observe therapeutic effect of electroacupuncture (EA) at Jiaji (EX-B 2) combined with laser needle-knife on lumbar disc herniation.

METHODSOne hundred and twenty cases of lumbar disc herniation were divided into an Jiaji EA group, a laser needle-knife group and a combination group (Jiaji EA plus laser needle-knife) according to random number table. Changes of symptoms at different stages before and after treatment were investigated with SF-MPQ cumulative scores.

RESULTSAll the 115 cases completed all of the study, SF-MPQ score in the combination group was significantly lower than those in the Jiaji EA group and the laser needle-knife group (P<0. 01, P<0.05), with a significant difference at the end of treatment of 2 weeks (P<0.01) between the laser needle-knife group and the Jiaji EA group, and with no significant difference at treatment of 1 week and 3 weeks (P>0.05). The recurrence rate at a half year later in the combination group was significantly lower than those in laser needle-knife group and the Jiaji EA group (P<0.05).

CONCLUSIONEA at Jiaji (EX-B 2) combined with laser needle-knife can significantly increase clinical therapeutic effect, alleviate pain of the patient and reduce recurrence.

Acupuncture Points ; Acupuncture Therapy ; methods ; Adult ; Aged ; Combined Modality Therapy ; Electroacupuncture ; Female ; Humans ; Intervertebral Disc Displacement ; therapy ; Lumbar Vertebrae ; Male ; Middle Aged

OBJECTIVETo investigate the influence of baicalin on the IL-1beta induced pro-MMP-1 in HGF and the effects of baicalin on MMP-3 expression in periodontal ligament cells (PDLCs).

METHODSThe amount of secreted pro-MMP-1 and MMP-3 expression was detected by ELISA and cell immunochemistry.

RESULTS(1) The amount of secreted pro-MMP-1 (3.333 +/- 0.123) microg/L increased significantly following 1 microg/L of IL-1beta, compared with control group (1.960 +/- 0.180) microg/L. Addition of baicalin to cell culture medium for 1 hour following IL-1beta decreased pro-MMP-1 secretion in a dose-dependent manner in the range of 10 approximately 1,000 microg/L. (2) 1 microg/L IL-1beta could significantly stimulate the synthesis and secretion of MMP-3 in PDLCs. (3) The baicalin could not interfere the synthesis of MMP-3, but could inhibit the release of MMP-3 from PDLCs.

CONCLUSIONSBaicalin could inhibit the secretion of pro-MMP-1 and MMP-3 expression in IL-1beta induced HGF and PDLCs, which suggests that baicalin may play an important role in preventing and treating periodontal disease.

Collagenases ; biosynthesis ; genetics ; Enzyme Precursors ; biosynthesis ; genetics ; Fibroblasts ; enzymology ; pathology ; Flavonoids ; pharmacology ; Gingiva ; enzymology ; pathology ; Humans ; Interleukin-1 ; pharmacology ; Interleukin-1beta ; Matrix Metalloproteinase 1 ; Metalloendopeptidases ; biosynthesis ; genetics ; Peptide Fragments ; pharmacology ; Periodontal Ligament ; enzymology ; pathology ; Periodontitis ; enzymology ; pathology ; Scutellaria ; chemistry